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Cited by 17Pluripotent Stem Cells ResearchHair Growth and DisordersTissue Engineering and Regenerative Medicine

Coordinated Development of Immune Cell Populations in Vascularized Skin Organoids from Human Induced Pluripotent Stem Cells

Open access
Mostina, Mitchell, Sun, Jane, Sim, Seen Ling, Ahmed, Imaan A., Souza‐Fonesca‐Guimaraes, Fernando, Wolvetang, Ernst J., Brown, Jason, Kumari, Snehlata +2 more·2025·Advanced Healthcare Materials
DOI 10.1002/adhm.202502108PMC PMC12683213
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Abstract

Human skin is a highly vascularized organ, where blood vessels perform essential roles in maintaining skin homeostasis and participate in thermal regulation. Currently, there is an unmet challenge to generate vascularized, functional skin models. In this study, human fetal placental endothelial colony forming cells (ECFC) are incorporated into human induced pluripotent stem cell (hiPSC)‐derived skin organoids (SKO), forming capillary‐like structures and generating endothelialized SKOs. However, this approach is limited by the inability of ECFCs to establish a complete vascular network, and impeding full epidermal stratification and hair follicle morphogenesis. In independent experiments, hiPSC‐derived vascular organoids (VO), are incorporated into the SKO, forming complex vascular structures and generating fully vascularized skin organoids (VSKO) with resident immune cell populations. Immunofluorescence microscopy and flow cytometric analyses reveal the transfer and integration of endothelial, mural, hematopoietic, and mesenchymal cells from VOs into the skin components of VSKOs. This study pioneers the establishment of VSKOs as a transformative platform for studying human skin biology, and immune‐skin interactions with applications in investigating inflammatory and other immune‐mediated skin disorders.

AI snapshot

Written from the paper’s full text.

TL;DR

The paper reports a hiPSC-derived vascularized skin organoid (VSKO) made by co-culturing one skin organoid with two hiPSC-derived vascular organoids on an air-liquid interface, yielding hair-bearing skin with perifollicular vessels and migrating immune cells. It contrasts this with an ECFC-based endothelialized skin organoid model that formed capillary-like structures but impaired hair follicle and keratinization programs.

Key findings

  • ECFC co-culture endothelialized SKOs: GFP-ECFCs formed CD31+/CD34+ capillary-like structures around SKOs, but the SKO + GFP-ECFC group lacked hair follicles and K10+ keratinization, unlike SKO Alone; GFR-M also prevented hair follicle development in SKO + Matrigel and SKO + GFP-ECFC groups.
  • hiPSC-derived VOs contained heterogeneous vascular cell populations: flow cytometry on day 42 showed that within the CD31−/CD45− population, ≈60% were CD90+/PDGFRβ+ and 36% were CD90+/PDGFRα+; VOs also had CD31+ endothelial, CD45+ hematopoietic, CD90+ mesenchymal, and PDGFRβ+ pericyte populations, plus DLL4+ arterial, EMCN+ venous, and α-SMA+ mural cells.
  • VSKOs generated from P111 and C32 hiPSC lines showed no necrotic cores at day 115, a thick K10+ epidermal layer, hair follicles, and CD34+/CD31+/NG2+ capillaries; GFP+ VO-derived vascular structures migrated into the dermis and perifollicular regions, with vascular-like structures reported as significantly greater than 1 mm in length.
  • VSKOs contained immune cells: CD45+ cells were distributed throughout the VSKO, predominantly in dermis, and all CD45+ cells were GFP+ when two GFP+ VOs were co-cultured with one GFP− SKO; CD68+ macrophages, CD207+ cells near hair follicles, and CD66b+ neutrophils were detected.

Methods

  • Two hiPSC lines, P111 and C32, were used to generate SKOs, VOs, and VSKOs. SKOs were differentiated as previously described; ECFCs were GFP-tagged, primed with increasing OMM/VEGF, and co-cultured with SKOs at day 18 in a GFR-M droplet until day 115.
  • VOs were differentiated from 3D hiPSC aggregates with Chir99021 and BMP-4 at day 0, SB431542 from day 2, and a GFR-M droplet at day 7; VOs were maintained in EGM2-microvascular medium then OMM with VEGF. VSKOs were made by placing one SKO with two VOs in a 30 µL GFR-M droplet on a Transwell air-liquid interface at day 18 of SKO differentiation, maintained to day 115.
  • Characterization used brightfield microscopy, immunofluorescence microscopy on cryosections and whole mounts, hematoxylin and eosin staining, and flow cytometric analysis of day 42 VOs for PDGFRβ, PDGFRα, CD31, CD45, and CD90.

Limitations

  • The paper states that ECFC-based endothelialized SKOs were limited by inability to establish a complete vascular network, impaired epidermal stratification and hair follicle morphogenesis, and an allogenic ECFC component that increases immune rejection risk for downstream grafting.
  • The paper states that further characterization is required to understand ECFC-SKO molecular interactions, that immune cell identity and function require further studies at extended time points, that the impact of vascularization on folliculogenesis needs further assessment, and that drug screening studies were not conducted.
  • Evident from the text: the work is in vitro and marker-based, with no functional immune challenge, transplantation, or graft integration experiments reported; immune populations are identified by immunofluorescence markers rather than functional assays.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1002/adhm.202502108

Cites

79 of 79 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

64 of 92 cited sources checked · 24 supported · 1 flagged · accuracy not yet measured

  • Interpretationcitation

    The increased cartilage and mesenchymal lineage formation in SKOs receiving ECFCs may be attributed to secretion of growth factors such as platelet-derived growth factor and transforming growth factor-β by ECFCs, which promote mesenchymal lineage differentiation.

    “that may be attributed to the secretion of growth factors such as platelet‐derived growth factor and transforming growth factor‐β by ECFCs, that promote mesenchymal lineage differentiation.[ 69 , 70 , 71 ]”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites [69] C. H. Lin, B. Lilly, Stem Cells Dev. 2014, 23, 2581. 24914692 PMC4201243 source

      Split into 2 atoms — 0 confirmed, 1 partially supported, 1 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [70] T. Liang, L. Zhu, W. Gao, M. Gong, J. Ren, H. Yao, K. Wang, D. Shi, FEBS Open Bio. 2017, 7, 1722. PMC5666384 29123981 source

      Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured
    • cites [71] F. Ng, S. Boucher, S. Koh, K. S. Sastry, L. Chase, U. Lakshmipathy, C. Choong, Z. Yang, M. C. Vemuri, M. S. Rao, V. Tanavde, Blood 2008, 112, 295. 18332228 source
  • Premisecitation

    There has been a complete absence of an endogenously differentiating vascularized, hair-bearing human skin equivalent that contains immune cell populations.

    “While many attempts to generate complex 3D skin equivalents have achieved vascularization, hair follicle generation, and immune cell co‐cultures, there has been a complete absence of endogenously differentiating a vascularized, hair‐bearing human skin equivalent that contains immune cell populations.[ 21 , 22 , 23 , 24 , 25 , 26 , 27 ]”
    Quote in the paperSubstantial claim4 of 7 sources checked
    • cites [21] S. Sun, L. Jin, Y. Zheng, J. Zhu, Nat. Commun. 2022, 13, 5481. 36123328 PMC9485166 source

      The Introduction (page unavailable) describes separate advances in vascularized skin models and models incorporating immune cells or dermal papilla cells. It does not address whether a human skin equivalent combining endogenous differentiation, vasculature, hair, and immune cell populations was entirely absent. A search for hair-bearing, endogenously differentiating models found no passage establishing that claim. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [22] D. M. Supp, A. P. Supp, S. M. Bell, S. T. Boyce, J. Invest. Dermatol. 2000, 114, 5. 10620108 source
    • cites [23] R. Rimal, S. Muduli, P. Desai, A. B. Marquez, M. Möller, I. Platzman, J. Spatz, S. Singh, Adv. Healthcare Mater. 2024, 13, 2303351. PMC11468127 38277705 source

      The Skin Organoids section describes hair-bearing models derived from human iPSCs but identifies vascularization and immune competence as challenges. Challenges and Outlook also notes a lack of immune cells and blood vessels in skin organoids, while Resemblance with Native Human Skin says a complete functional model is lacking. These passages support the combination as an unmet challenge, but do not establish the categorical claim that no endogenously differentiating model with all three features exists. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [24] E. Hofmann, A. Schwarz, J. Fink, L. P. Kamolz, P. Kotzbeck, Biomedicines 2023, 11, 794. 36979772 PMC10045055 source

      Sections 4.1, 4.2, and 4.5 describe separate advances in immune-cell incorporation, vascularization, and hair-follicle formation, while section 5 says current models still face substantial limitations. The paper does not establish the claimed complete absence of a single endogenously differentiating human skin equivalent combining vascularization, hair, and immune-cell populations. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [25] A. Shafiee, J. Sun, I. A. Ahmed, F. Phua, G. R. Rossi, C.‐Y. Lin, F. Souza‐Fonseca‐Guimaraes, E. J. Wolvetang, J. Brown, K. Khosrotehrani, Small 2024, 20, 2304879. 38044307 source
    • cites [26] J. Lee, C. C. Rabbani, H. Gao, M. R. Steinhart, B. M. Woodruff, Z. E. Pflum, A. Kim, S. Heller, Y. Liu, T. Z. Shipchandler, K. R. Koehler, Nature 2020, 582, 399. 32494013 PMC7593871 source

      The paper reports hair-bearing human skin organoids but says it did not detect progenitors of Langerhans cells, which are resident skin immune cells (section labelled Abstract, page unavailable). It also reports that vasculature grew into the organoids only after grafting into mice. These findings support a gap in this model, but the paper does not establish the claimed complete absence of any endogenously differentiating human skin equivalent combining all three features. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [27] D. Marino, J. Luginbuhl, S. Scola, M. Meuli, E. Reichmann, Sci. Transl. Med. 2014, 6, 221. 24477001 source
  • Premisecitation

    The physiological relevance of hiPSC-derived SKOs remains limited by the absence of vascular and immune components, as SKOs are derived from differentiation of the surface ectoderm.

    “Despite their experimental advantages over existing skin equivalents, the physiological relevance of hiPSC‐derived SKOs remains limited by the absence of vascular and immune components, as SKOs are derived from differentiation of the surface ectoderm.[ 36 , 37 , 38 ]”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites [36] J. Lee, K. R. Koehler, Exp. Dermatol. 2021, 30, 613. 33507537 PMC8265774 source

      The section “Increasing Complexity: Skin Organoids with Vasculature, Immune Cells, and Sweat Glands” says skin organoids lack endothelial cells, pericytes, and immune cells, supporting the stated limitation. But “The Challenge of Growing Skin Appendages in a Dish” says the organoids are generated by co-inducing surface ectoderm and cranial neural crest cells, not by differentiating surface ectoderm alone. The proposed explanation for the missing components is therefore not supported as stated. No page numbers are available in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [37] Z. X. Hong, S. T. Zhu, H. Li, J. Z. Luo, Y. Yang, Y. An, X. Wang, K. Wang, Mil. Med. Res. 2023, 10, 40. 37605220 PMC10463602 source

      The Limitations section (page unavailable) identifies incomplete vasculature and difficulty establishing immune cell clusters as limitations, and says missing immune components may compromise the relevance of hiPSC-derived skin organoid models. But the Hair follicle organoid section describes co-induction of surface ectoderm and cranial neural crest cells; the paper does not attribute the vascular and immune limitations to differentiation from surface ectoderm alone. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [38] M. Hosseini, K. R. Koehler, A. Shafiee, Adv. Healthcare Mater. 2022, 11, 2201626. PMC11469047 36063498 source

      The Conclusions and Outlook section says human PSC-derived skin organoids differ from skin in structure and function and that current skin models lack a vascular network. Figure 4 describes coinduction of surface ectoderm and cranial neural crest cells, rather than derivation from surface ectoderm alone. The cited passages do not establish that these organoids lack immune components. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
  • Premisecitation

    The air-liquid interface has been shown to enhance differentiation of hiPSC-derived SKOs.

    “Exposing the SKO to air recapitulates the external environment human skin is exposed to and has shown to enhance differentiation of hiPSC‐derived SKOs.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] J. Sun, I. Ahmed, J. Brown, K. Khosrotehrani, A. Shafiee, Burns Trauma 2025, 13, 070. PMC11736897 39822647 source
    • Rests on an assumption: Exposing the SKO to air recapitulates the external environment human skin is exposed to.
  • Findingcitation

    Hair follicles in VSKOs are SOX2+, K14+, K15+, and K17+, as seen in the current SKO model.

    “Figure 7c also shows that the hair follicles in VSKOs are SRY‐box transcription factor 2 (SOX2)+, K14+, cytokeratin 15 (K15)+, and K17+, as seen in the current SKO model.[ 25 , 26 ]”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [25] A. Shafiee, J. Sun, I. A. Ahmed, F. Phua, G. R. Rossi, C.‐Y. Lin, F. Souza‐Fonseca‐Guimaraes, E. J. Wolvetang, J. Brown, K. Khosrotehrani, Small 2024, 20, 2304879. 38044307 source
    • cites [26] J. Lee, C. C. Rabbani, H. Gao, M. R. Steinhart, B. M. Woodruff, Z. E. Pflum, A. Kim, S. Heller, Y. Liu, T. Z. Shipchandler, K. R. Koehler, Nature 2020, 582, 399. 32494013 PMC7593871 source

      The paper reports SOX2+ dermal papilla clusters in organoid hair follicles (Abstract, Fig. 1i–l) and KRT15+ stem-cell-like cells in the presumptive bulge of xenografted follicles (Abstract, Fig. 4j). Our copy does not establish K14+ or K17+ hair follicles, nor does this paper examine the citing authors’ VSKOs. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
  • Classified as probably not a substantial claim. 18
  • May combine two claims. 47
  • The quote only partly states this claim. 1

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    hiPSC-derived skin organoids have hair follicles, pigmentation, dermal stratification, appendage patterning, and innervation.

    “Recent advances in human induced pluripotent stem cell (hiPSC) technology have enabled the generation of skin organoids (SKO) with hair follicles, pigmentation, dermal stratification, appendage patterning, and innervation.[ 25 , 26 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 ]”
    Quote in the paperNot a substantial claim9 of 10 sources checked
    • cites [25] A. Shafiee, J. Sun, I. A. Ahmed, F. Phua, G. R. Rossi, C.‐Y. Lin, F. Souza‐Fonseca‐Guimaraes, E. J. Wolvetang, J. Brown, K. Khosrotehrani, Small 2024, 20, 2304879. 38044307 source
    • cites [26] J. Lee, C. C. Rabbani, H. Gao, M. R. Steinhart, B. M. Woodruff, Z. E. Pflum, A. Kim, S. Heller, Y. Liu, T. Z. Shipchandler, K. R. Koehler, Nature 2020, 582, 399. 32494013 PMC7593871 source

      The paper reports hair follicles, pigmentation, periodically spaced appendages, and innervation in skin organoids; it also reports hair-bearing organoids from a hiPSC line (Abstract; no page available). But it describes a stratified epidermis and a fat-rich dermis, not dermal stratification. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [28] S. Y. Jung, H. J. You, M. J. Kim, G. Ko, S. Lee, K. S. Kang, iScience 2022, 25, 105150. 36193049 PMC9526179 source
    • cites [29] P. Li, S. T. Pachis, G. Xu, R. Schraauwen, R. Incitti, A. C. de Vries, M. J. Bruno, M. P. Peppelenbosch, I. Alam, K. Raymond, Q. Pan, Nat. Microbiol. 2023, 8, 2067. 37828248 source

      The paper’s introductory section describes hiPSC-derived skin organoids with multilayer structure, hair follicles, and sebaceous glands; Skin organoid generation also describes producing hair-bearing organoids. Our copy does not establish pigmentation, appendage patterning, or innervation, so it supports only part of the cited list. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [30] J. Ma, J. liu, D. Gao, X. Li, Q. Zhang, L. Lv, Y. Wang, J. Li, Y. Zhu, Z. Wu, H. Hu, Y. Li, L. Ma, Q. Liu, Z. Hu, S. Zhang, Y. Zhou, M. Wang, L. Leng, Adv. Sci. 2022, 9, 2104192. PMC8895131 34971128 source

      The Results (‘Formation of Skin Organoids with Various hiPSC-Derived Skin Cells’; page unavailable) report mature hair follicles and a stratified epithelium. The Results (‘SARS-CoV-2 Can Infect the Nervous System of Skin’; page unavailable) also show a neuronal network around the follicles. But the paper identifies melanosome-related proteins rather than demonstrating visible pigmentation, and it does not establish dermal stratification or appendage patterning; epithelial stratification is not dermal stratification. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [31] V. Ramovs, H. Janssen, I. Fuentes, A. Pitaval, W. Rachidi, S. M. Chuva de Sousa Lopes, C. Freund, X. Gidrol, C. L. Mummery, K. Raymond, Stem Cell Rep. 2022, 17, 1279. PMC9213820 35561682 source
    • cites [32] I. Ahmed, J. Sun, J. Brown, K. Khosrotehrani, A. Shafiee, Biol. Methods Protoc. 2024, 9, 019. PMC11009018 38605978 source

      The Introduction and the Methods figure 4 caption describe hiPSC-derived skin organoids with hair follicles, other appendages, hair placodes, and neurites. Troubleshooting, Problem 4 says follicles can develop pigmentation. The copy describes epidermal layers and an epidermis–dermis arrangement, but does not establish dermal stratification or a defined pattern of appendages; pigmentation is described as conditional. Page numbers were not available in our copy. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [33] J. Sun, I. Ahmed, J. Brown, K. Khosrotehrani, A. Shafiee, Burns Trauma 2025, 13, 070. PMC11736897 39822647 source

      The Results sections (pages not specified in our copy) document hair follicles, pigmentation, sebaceous and sweat-gland structures, and innervation in hiPSC-derived skin organoids. They describe a multilayered epidermis and appendages adjacent to hair follicles, but do not establish dermal stratification or a distinct patterning of appendages. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [34] I. A. Ahmed, J. Sun, M. J. Kong, K. Khosrotehrani, A. Shafiee, Exp. Dermatol. 2024, 33, 70017. 39582396 source

      Figure 1 (Conclusions & Future Perspectives; page not provided) shows hiPSC-derived skin organoids developing hair placodes and hair follicles, supporting the hair-follicle portion of the claim. The copy does not establish pigmentation, dermal stratification, broader appendage patterning, or innervation; searches for those features found no supporting passage. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [35] N. H. Gopee, E. Winheim, B. Olabi, C. Admane, A. R. Foster, N. Huang, R. A. Botting, F. Torabi, D. Sumanaweera, A. P. Le, J. Kim, L. Verger, E. Stephenson, D. Adão, C. Ganier, K. Y. Gim, S. A. Serdy, C. Deakin, I. Goh, L. Steele, K. Annusver, M.‐U. Miah, W. M. Tun, P. Moghimi, K. A. Kwakwa, T. Li, D. Basurto Lozada, B. Rumney, C. L. Tudor, K. Roberts, et al., Nature 2024, 635, 679. 39415002 PMC11578897 source

      The Single-cell atlas of human prenatal skin section (page not supplied) says the stem-cell-derived skin organoid model recapitulates prenatal hair-follicle components, neural cells and dermal fibroblasts, and contains melanocytes. The paper does not establish pigmentation, dermal stratification, appendage patterning or innervation as organoid features; neural cells and melanocytes alone do not demonstrate innervation or pigmentation. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    ECFCs sourced from cord blood or placenta can create de novo vascular networks in vitro and maintain the capacity to integrate with host vasculature in vivo.

    “Endothelial colony forming cells (ECFC), sourced from cord‐blood or the placenta, demonstrate the ability to create de novo vascular networks in vitro, whilst maintaining the capacity to integrate with host vasculature in vivo.[ 46 , 47 , 48 ]”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites [46] C. L. O'Neill, K. J. McLoughlin, S. E. J. Chambers, J. Guduric‐Fuchs, A. W. Stitt, R. J. Medina, Front. Med. 2018, 5, 273. PMC6232760 30460233 source
    • cites [47] K. Gao, S. He, P. Kumar, D. Farmer, J. Zhou, A. Wang, World J. Stem Cells 2020, 12, 123. 32184937 PMC7062038 source

      The Results section ('CV-ECFCs are functionally comparable to CB-ECFCs'; page unavailable) reports that placenta- and cord-blood-derived ECFCs formed similar tubular structures in vitro. The Introduction says ECFCs can integrate into the circulatory system, but this paper does not demonstrate that its placental ECFCs integrate with host vasculature in vivo. That part of the claim is therefore not established for both cell sources. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [48] M. J. Javed, L. E. Mead, D. Prater, W. K. Bessler, D. Foster, J. Case, W. S. Goebel, M. C. Yoder, L. S. Haneline, D. A. Ingram, Pediatr. Res. 2008, 64, 68. 18360305 source

  • May combine two claims.

    Premisecitation

    Although macrophage co-culture with SKOs has been reported, a physiologically relevant vascularized hiPSC-derived SKO has yet to be achieved.

    “Although macrophage co‐culture with SKOs has been reported,[ 35 ] a physiologically relevant vascularized hiPSC‐derived SKO has yet to be achieved.[ 36 , 37 , 38 ]”
    Quote in the paperSubstantial claim4 of 4 sources checked
    • cites [35] N. H. Gopee, E. Winheim, B. Olabi, C. Admane, A. R. Foster, N. Huang, R. A. Botting, F. Torabi, D. Sumanaweera, A. P. Le, J. Kim, L. Verger, E. Stephenson, D. Adão, C. Ganier, K. Y. Gim, S. A. Serdy, C. Deakin, I. Goh, L. Steele, K. Annusver, M.‐U. Miah, W. M. Tun, P. Moghimi, K. A. Kwakwa, T. Li, D. Basurto Lozada, B. Rumney, C. L. Tudor, K. Roberts, et al., Nature 2024, 635, 679. 39415002 PMC11578897 source
    • cites [36] J. Lee, K. R. Koehler, Exp. Dermatol. 2021, 30, 613. 33507537 PMC8265774 source
    • cites [37] Z. X. Hong, S. T. Zhu, H. Li, J. Z. Luo, Y. Yang, Y. An, X. Wang, K. Wang, Mil. Med. Res. 2023, 10, 40. 37605220 PMC10463602 source
    • cites [38] M. Hosseini, K. R. Koehler, A. Shafiee, Adv. Healthcare Mater. 2022, 11, 2201626. PMC11469047 36063498 source

  • May combine two claims.

    Premisecitation

    The ALI system improved SKO development by sustaining hair follicle development and enhanced keratinocyte stratification, compared to floating culture.

    “The ALI system improved SKO development by sustaining hair follicle development and enhanced keratinocyte stratification, when compared to floating culture.[ 33 ]”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] J. Sun, I. Ahmed, J. Brown, K. Khosrotehrani, A. Shafiee, Burns Trauma 2025, 13, 070. PMC11736897 39822647 source

  • May combine two claims.

    Findingown data

    H&E staining demonstrated formation of a thick epithelium, hair follicles, and dermal papillae within the dermis in VSKOs, consistent with previous reports on SKOs.

    “Furthermore, hematoxylin and eosin staining demonstrated the formation of a thick epithelium, hair follicles, and dermal papillae within the dermis, consistent with previous reports on SKOs (Figure 7a).[ 32 ]”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [32] I. Ahmed, J. Sun, J. Brown, K. Khosrotehrani, A. Shafiee, Biol. Methods Protoc. 2024, 9, 019. PMC11009018 38605978 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    During human development, skin is derived from the surface ectoderm, while blood vessels and immune cells are both derived from the mesoderm.

    “During human development, skin is derived from the surface ectoderm, while blood vessels and the immune cells are both derived from the mesoderm.[ 51 , 60 , 61 ]”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [51] R. A. Wimmer, A. Leopoldi, M. Aichinger, D. Kerjaschki, J. M. Penninger, Nat. Protoc. 2019, 14, 85073082. 31554955 source
    • cites [60] X. Gao, C. Xu, N. Asada, P. S. Frenette, Development 2018, 145, 139691. PMC5825844 29358215 source

      The Introduction states that the hematopoietic system is of mesodermal origin and describes the mouse embryonic region containing the dorsal aorta as mesoderm-derived. It does not address skin’s origin from surface ectoderm, and its vessel-development example is in mice rather than humans. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [61] W. D. Tucker, Y. Arora, M. K. Anatomy, Blood Vessels, StatPearls, Treasure Island (FL) 2024.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Premisecitation

    Previous human SKO models were devoid of vascularization and immune cells.

    “Previous human SKO models were devoid of vascularization and immune cells.[ 25 , 26 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 59 ]”
    Quote in the paperSubstantial claim9 of 10 sources checked
    • cites [25] A. Shafiee, J. Sun, I. A. Ahmed, F. Phua, G. R. Rossi, C.‐Y. Lin, F. Souza‐Fonseca‐Guimaraes, E. J. Wolvetang, J. Brown, K. Khosrotehrani, Small 2024, 20, 2304879. 38044307 source
    • cites [26] J. Lee, C. C. Rabbani, H. Gao, M. R. Steinhart, B. M. Woodruff, Z. E. Pflum, A. Kim, S. Heller, Y. Liu, T. Z. Shipchandler, K. R. Koehler, Nature 2020, 582, 399. 32494013 PMC7593871 source

      The organoid-characterization passage (indexed as Abstract; page unavailable) says the authors did not detect progenitors of Langerhans cells, a type of skin immune cell. The grafting passage (also indexed as Abstract; page unavailable) reports that vasculature grew into organoids after transplantation. These findings support limitations of this model, but do not establish that all previous human skin organoid models lacked vascularization and immune cells. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [28] S. Y. Jung, H. J. You, M. J. Kim, G. Ko, S. Lee, K. S. Kang, iScience 2022, 25, 105150. 36193049 PMC9526179 source
    • cites [29] P. Li, S. T. Pachis, G. Xu, R. Schraauwen, R. Incitti, A. C. de Vries, M. J. Bruno, M. P. Peppelenbosch, I. Alam, K. Raymond, Q. Pan, Nat. Microbiol. 2023, 8, 2067. 37828248 source

      The Discussion states that immune cells are currently lacking in the authors’ skin organoids, supporting that part of the claim. Our copy does not establish that these or previous human skin organoid models lacked vascularization; a search for vascularization and blood vessels found no supporting passage. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [30] J. Ma, J. liu, D. Gao, X. Li, Q. Zhang, L. Lv, Y. Wang, J. Li, Y. Zhu, Z. Wu, H. Hu, Y. Li, L. Ma, Q. Liu, Z. Hu, S. Zhang, Y. Zhou, M. Wang, L. Leng, Adv. Sci. 2022, 9, 2104192. PMC8895131 34971128 source

      The Results section describes this skin organoid’s cell types and reports proteins associated with vascular-related cells, but does not establish whether it has vascularization. Neither the supplied passages nor searches for vascularization and immune cells establish that this model—or previous human skin organoid models—lacked both features. Page numbers were not available in our copy. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [31] V. Ramovs, H. Janssen, I. Fuentes, A. Pitaval, W. Rachidi, S. M. Chuva de Sousa Lopes, C. Freund, X. Gidrol, C. L. Mummery, K. Raymond, Stem Cell Rep. 2022, 17, 1279. PMC9213820 35561682 source

      The Introduction and Results and discussion describe the organoids’ epidermal-dermal junction and cell types, but do not state whether previous skin organoid models lacked vascularization or immune cells. Searches for vascularization, endothelial cells, and immune cells found no passage addressing both claimed absences; a reference to endothelialized skin equivalents does not establish them. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [32] I. Ahmed, J. Sun, J. Brown, K. Khosrotehrani, A. Shafiee, Biol. Methods Protoc. 2024, 9, 019. PMC11009018 38605978 source
    • cites [33] J. Sun, I. Ahmed, J. Brown, K. Khosrotehrani, A. Shafiee, Burns Trauma 2025, 13, 070. PMC11736897 39822647 source
    • cites [34] I. A. Ahmed, J. Sun, M. J. Kong, K. Khosrotehrani, A. Shafiee, Exp. Dermatol. 2024, 33, 70017. 39582396 source

      The Conclusions & Future Perspectives section says that current skin substitutes lack the complexity of human skin, but does not say that previous SKO models lacked vascularization or immune cells. The Results describe SKO dermis and hair follicles; searches for vascularization, vasculature, blood vessels, and immune cells found no passage addressing the claimed absence. (LLM, conf=0.75)

      Experimental — accuracy not yet measured
    • cites [35] N. H. Gopee, E. Winheim, B. Olabi, C. Admane, A. R. Foster, N. Huang, R. A. Botting, F. Torabi, D. Sumanaweera, A. P. Le, J. Kim, L. Verger, E. Stephenson, D. Adão, C. Ganier, K. Y. Gim, S. A. Serdy, C. Deakin, I. Goh, L. Steele, K. Annusver, M.‐U. Miah, W. M. Tun, P. Moghimi, K. A. Kwakwa, T. Li, D. Basurto Lozada, B. Rumney, C. L. Tudor, K. Roberts, et al., Nature 2024, 635, 679. 39415002 PMC11578897 source

      The Single-cell atlas of human prenatal skin section says the SkO model lacked immune cells, supporting that part of the claim. It says endothelial cells were markedly reduced, not absent; the Macrophages support prenatal skin angiogenesis section also describes SkO endothelial cells. The source therefore does not support saying these models were devoid of vascularization. (LLM, conf=0.91)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    Previous reports described the absence of immune and vascular components during hiPSC-derived SKO differentiation.

    “Previous reports described the absence of immune and vascular components during hiPSC‐derived SKO differentiation.[ 25 , 26 , 33 ]”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [25] A. Shafiee, J. Sun, I. A. Ahmed, F. Phua, G. R. Rossi, C.‐Y. Lin, F. Souza‐Fonseca‐Guimaraes, E. J. Wolvetang, J. Brown, K. Khosrotehrani, Small 2024, 20, 2304879. 38044307 source
    • cites [26] J. Lee, C. C. Rabbani, H. Gao, M. R. Steinhart, B. M. Woodruff, Z. E. Pflum, A. Kim, S. Heller, Y. Liu, T. Z. Shipchandler, K. R. Koehler, Nature 2020, 582, 399. 32494013 PMC7593871 source

      In the organoid cell-lineage analysis (section labelled Abstract; page unavailable), the paper says it did not detect progenitors of Langerhans cells, which are skin immune cells. It does not explicitly report an absence of vascular components during differentiation; it reports that vasculature grew into organoids after grafting into mice (section labelled Abstract; page unavailable). The claim therefore extends beyond what the paper establishes. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [33] J. Sun, I. Ahmed, J. Brown, K. Khosrotehrani, A. Shafiee, Burns Trauma 2025, 13, 070. PMC11736897 39822647 source

  • Classified as probably not a substantial claim.

    Premisecitation

    Vasculogenic potential of endothelial progenitors is enhanced when combined with a mesenchymal component.

    “Previous studies have shown that the vasculogenic potential of endothelial progenitors is enhanced when combined with a mesenchymal component.[ 64 , 73 ]”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [64] A. Shafiee, J. Patel, J. S. Lee, D. W. Hutmacher, N. M. Fisk, K. Khosrotehrani, Sci. Rep. 2017, 7, 13558. 29051567 PMC5648925 source
    • cites [73] A. Shafiee, J. Patel, H. Yi Wong, P. Donovan, D. W. Hutmacher, N. M. Fisk, K. Khosrotehrani, FASEB J. 2017, 31, 006619610. 28045376 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Endothelial cells have been used to endothelialize different organoids.

    “Endothelial cells have been used to endothelialize different organoids.[ 67 , 68 ]”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [67] J. C. Maggiore, R. LeGraw, A. Przepiorski, J. Velazquez, C. Chaney, T. Vanichapol, E. Streeter, Z. Almuallim, A. Oda, T. Chiba, A. Silva‐Barbosa, J. Franks, J. Hislop, A. Hill, H. Wu, K. Pfister, S. E. Howden, S. C. Watkins, M. H. Little, B. D. Humphreys, S. Kiani, A. Watson, D. B. Stolz, A. J. Davidson, T. Carroll, O. Cleaver, S. Sims‐Lucas, M. R. Ebrahimkhani, N. A. Hukriede, Kidney Int. 2024, 106, 1086. 38901605 PMC11912416 source
    • cites [68] A.‐L. Ament, M. Heiner, M. C. Hessler, I. Alexopoulos, K. Steeg, U. Gärtner, A. I. Vazquez‐Armendariz, S. Herold, Am. J. Respir. Cell Mol. Biol. 2025, 72, 124. 39226154 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Langerhans cells are a unique cell population to the skin organ, where fate-mapping of macrophages demonstrates in situ proliferation to Langerhans cells in the skin during embryonic development.

    “Langerhans cells are a unique cell population to the skin organ, where fate‐mapping of macrophages demonstrates in situ proliferation to Langerhans cells in the skin during embryonic development.[ 78 , 79 , 80 , 81 ]”
    Quote in the paperNot a substantial claim3 of 4 sources checked
    • cites [78] M. Collin, P. Milne, Curr. Opin. Hematol. 2016, 23, 28. 26554892 PMC4685746 source

      The section “A primitive origin of LCs from yolk sac macrophages” says fate-mapping experiments showed primitive macrophages proliferating in situ and giving rise to Langerhans cells. But the Introduction says Langerhans cells also reside in the corneal, buccal, gingival and genital mucosae, so they are not unique to the skin. (LLM, conf=0.90)

      Experimental — accuracy not yet measured
    • cites [79] F. Ginhoux, S. Jung, Nat. Rev. Immunol. 2014, 14, 392. 24854589 source
    • cites [80] Y. Wang, K. J. Szretter, W. Vermi, S. Gilfillan, C. Rossini, M. Cella, A. D. Barrow, M. S. Diamond, M. Colonna, Nat. Immunol. 2012, 13, 753. 22729249 PMC3941469 source

      The Introduction identifies Langerhans cells as resident cells of the epidermis, and the Results section 'The development of microglia and LCs require IL-34' says their primitive precursors populate the skin before birth and expand around birth (page unavailable). The paper does not report macrophage fate-mapping or demonstrate that macrophages proliferate in situ into Langerhans cells during embryonic development. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [81] M. Greter, I. Lelios, P. Pelczar, G. Hoeffel, J. Price, M. Leboeuf, T. M. Kündig, K. Frei, F. Ginhoux, M. Merad, B. Becher, Immunity 2012, 37, 1050. 23177320 PMC4291117 source

      The Results section, “IL-34 Controls the Embryonic Development of LCs but Not Microglia” (page unavailable), reports that precursors seed the skin before birth and that LC differentiation occurs locally during embryonic development. It does not report fate-mapping evidence of in situ proliferation during embryogenesis; the paper discusses local self-renewal in adult LCs instead. It also says adult LCs derive predominantly from fetal liver monocytes, with a smaller contribution from yolk sac macrophages. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Interpretationcitation

    On day 115 of differentiation (≈16-week of embryonic development), the presence of CD207+ immune cells in the dermal region suggests two possibilities: migration of Langerhans cell precursors from the dermis, or presence of dermal dendritic cells, both of which express CD207+.

    “On day 115 of differentiation (≈16‐week of embryonic development), the presence of CD207+ immune cells in the dermal region suggests two possibilities: first, the migration of Langerhans cell precursors from the dermis[ 82 , 83 ]; and second, the presence of dermal dendritic cells,[ 82 , 83 ] both of which express CD207+.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [82] G. Hoeffel, Y. Wang, M. Greter, P. See, P. Teo, B. Malleret, M. Leboeuf, D. Low, G. Oller, F. Almeida, S. H. Y. Choy, M. Grisotto, L. Renia, S. J. Conway, E. R. Stanley, J. K. Y. Chan, L. G. Ng, I. M. Samokhvalov, M. Merad, F. Ginhoux, J. Exp. Med. 2012, 209, 1167. 22565823 PMC3371735 source

      Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured
    • cites [83] M. Haniffa, M. Gunawan, L. Jardine, J. Dermatol. Sci. 2015, 77, 85. 25301671 PMC4728191 source

      Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured
    • Rests on an assumption: Day 115 of differentiation corresponds to approximately 16 weeks of embryonic development.

  • May combine two claims.

    Interpretationcitation

    VSKOs can be used as a tool to investigate the interaction of hair follicles with vasculature, as well as hair-related pathologies.

    “VSKOs can also be used as a tool to investigate the interaction of hair follicles with vasculature, as well as hair‐related pathologies.[ 75 , 76 ]”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [75] K. Yano, L. F. Brown, M. Detmar, J. Clin. Invest. 2001, 107, 409. 11181640 PMC199257 source

      The Introduction and Discussion (page unavailable) show that perifollicular blood vessels affect hair growth and discuss a possible role for vascularization in hair-loss disorders. The study used mice and isolated vibrissa follicles, not VSKOs, so it does not establish that VSKOs can be used to investigate those interactions or pathologies. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [76] C. K. Goldman, J. C. Tsai, L. Soroceanu, G. Y. Gillespie, J. Invest. Dermatol. 1995, 104, 18S 7738377 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Mature human SKOs resemble human fetal craniofacial skin at 18-week of gestation.

    “Notably, this observation aligns with previous studies that mature human SKOs resemble human fetal craniofacial skin at 18‐week of gestation.[ 26 , 78 ]”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [26] J. Lee, C. C. Rabbani, H. Gao, M. R. Steinhart, B. M. Woodruff, Z. E. Pflum, A. Kim, S. Heller, Y. Liu, T. Z. Shipchandler, K. R. Koehler, Nature 2020, 582, 399. 32494013 PMC7593871 source
    • cites [78] M. Collin, P. Milne, Curr. Opin. Hematol. 2016, 23, 28. 26554892 PMC4685746 source

      The ‘Bone marrow independence of human LCs’ section says human Langerhans cells are fully formed at around 18–24 weeks of gestation. It does not compare mature human SKOs with fetal craniofacial skin. Searches for skin organoids, craniofacial skin, and the 18-week comparison found no passage addressing that claim; page numbers were not available in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Findingcitation

    SKOs appeared to maintain hair follicle development after vascularization, with perifollicular vascularization consistently observed throughout the VSKO as seen in vivo.

    “Whilst the VSKOs were co‐cultured on an ALI to recapitulate the external environment human skin is exposed to, SKOs appeared to maintain hair follicle development after vascularization, where perifollicular vascularization was consistently observed throughout the VSKO as seen in vivo.[ 75 ]”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [75] K. Yano, L. F. Brown, M. Detmar, J. Clin. Invest. 2001, 107, 409. 11181640 PMC199257 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    ALI culture, facilitated using Transwell system, has previously been shown to enhance hair follicle development in SKOs.

    “The ALI culture, facilitated using Transwell system, has previously shown to enhance hair follicle development in SKOs.[ 33 ]”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] J. Sun, I. Ahmed, J. Brown, K. Khosrotehrani, A. Shafiee, Burns Trauma 2025, 13, 070. PMC11736897 39822647 source

  • May combine two claims.

    Premisecitation

    Currently, SKOs lack vasculature and immune cells in a physiologically relevant context.

    “Currently, SKOs lack vasculature and immune cells in a physiologically relevant context.[ 25 , 35 , 36 , 37 , 38 ]”
    Quote in the paperSubstantial claim4 of 5 sources checked
    • cites [25] A. Shafiee, J. Sun, I. A. Ahmed, F. Phua, G. R. Rossi, C.‐Y. Lin, F. Souza‐Fonseca‐Guimaraes, E. J. Wolvetang, J. Brown, K. Khosrotehrani, Small 2024, 20, 2304879. 38044307 source
    • cites [35] N. H. Gopee, E. Winheim, B. Olabi, C. Admane, A. R. Foster, N. Huang, R. A. Botting, F. Torabi, D. Sumanaweera, A. P. Le, J. Kim, L. Verger, E. Stephenson, D. Adão, C. Ganier, K. Y. Gim, S. A. Serdy, C. Deakin, I. Goh, L. Steele, K. Annusver, M.‐U. Miah, W. M. Tun, P. Moghimi, K. A. Kwakwa, T. Li, D. Basurto Lozada, B. Rumney, C. L. Tudor, K. Roberts, et al., Nature 2024, 635, 679. 39415002 PMC11578897 source
    • cites [36] J. Lee, K. R. Koehler, Exp. Dermatol. 2021, 30, 613. 33507537 PMC8265774 source
    • cites [37] Z. X. Hong, S. T. Zhu, H. Li, J. Z. Luo, Y. Yang, Y. An, X. Wang, K. Wang, Mil. Med. Res. 2023, 10, 40. 37605220 PMC10463602 source
    • cites [38] M. Hosseini, K. R. Koehler, A. Shafiee, Adv. Healthcare Mater. 2022, 11, 2201626. PMC11469047 36063498 source

      The Conclusions and Outlook section says that current skin models lack a vascular network and specifically discusses this limitation after describing human skin organoids. Our copy does not say that skin organoids lack immune cells in a physiologically relevant context, so that part of the claim is unsupported by this source. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Macrophage recruitment to the skin begins during primitive hematopoiesis in the yolk sac, where they mature into skin-resident cells.

    “This observation is consistent with the fact that macrophage recruitment to the skin begins during primitive hematopoiesis in the yolk sac, where they mature into skin‐resident cells.[ 77 ]”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [77] O. Bhattacharjee, U. Ayyangar, A. S. Kurbet, V. Lakshmanan, D. Palakodeti, F. Ginhoux, S. Raghavan, Front. Immunol. 2021, 12, 718005. 34721382 PMC8553113 source

  • May combine two claims.

    Findingown data

    Compared to the SKO + Matrigel control, the GFR-M droplet containing ECFCs impacted the cyst-like structure of the SKO, with some endothelialized SKOs containing an NG2+ cartilaginous structure in the center.

    “However, compared to the SKO + Matrigel control group, the GFR‐M droplet containing ECFCs impacted the cyst‐like structure of the SKO, where heterogeneity was observed with some endothelialized SKOs containing a neural/glial antigen 2 (NG2)+ cartilaginous structure in the center (Figure 2a).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    hiPSC-derived VOs incorporated into SKO form complex vascular structures and generate fully vascularized skin organoids with resident immune cell populations.

    “In independent experiments, hiPSC‐derived vascular organoids (VO), are incorporated into the SKO, forming complex vascular structures and generating fully vascularized skin organoids (VSKO) with resident immune cell populations.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CD31+ and NG2+ cells co-localized and penetrated toward the center of the VO with the GFR-M droplet, indicating formation of vascular structures.

    “Also, CD31+ and NG2+ cells were co‐localized and penetrated toward the center of the VO with the GFR‐M droplet indicating formation of vascular structures (Figure 3e).”
    Quote in the paperSubstantial claim

  • May combine two claims. The quote only partly states this claim.

    Findingnone given

    ECFCs and GFR-M impede full epidermal stratification and hair follicle morphogenesis in SKOs.

    “However, this approach is limited by the inability of ECFCs to establish a complete vascular network, and impeding full epidermal stratification and hair follicle morphogenesis.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    At day 7, VOs exhibited CD31+ and NG2+ cells around the periphery, with PDGFRα homogenously expressed throughout the center.

    “At day 7, VOs exhibited the presence of CD31+ and NG2+ cells around the periphery, with platelet‐derived growth factor receptor α (PDGFRα) homogenously expressed throughout the center (Figure 3e).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    ECFCs incorporated into hiPSC-derived SKOs form capillary-like structures and generate endothelialized SKOs.

    “In this study, human fetal placental endothelial colony forming cells (ECFC) are incorporated into human induced pluripotent stem cell (hiPSC)‐derived skin organoids (SKO), forming capillary‐like structures and generating endothelialized SKOs.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    There is a heterogeneous population of CD34+ and CD34− cells within VSKOs, with vascular-like structures exclusively formed by GFP+/CD34+ cells.

    “There is a heterogeneous population of CD34⁺ and CD34− cells within VSKOs, with vascular‐like structures exclusively formed by GFP⁺/CD34⁺ cells, indicating the contribution of VOs to vascularization (Figure 7b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Most VO cells are CD90+ cells, which could be PDGFRα+ or β+.

    “We found most of the VO cells are CD90+ cells, which could be PDGFRα+ or β+.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Endothelial differentiation occurred in VSKOs, with DLL4+ arterial capillaries and EMCN+ venous capillaries observed.

    “Additionally, endothelial differentiation occurred, where DLL4+ arterial capillaries and EMCN+ venous capillaries were observed (Figure 6b,c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    VOs demonstrated a heterogeneous phenotype of endothelial cells (CD31+), hematopoietic cells (CD45+), mesenchymal cells (CD90+), and pericytes (PDGFRβ+).

    “VOs demonstrated a heterogenous phenotype of endothelial cells (CD31+), hematopoietic cells (CD45+), mesenchymal cells (CD90+), and pericytes (platelet‐derived growth factor receptor β (PDGFRβ+)) (Figure 4a–c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Within the CD31−/CD45− population, approximately 60% were CD90+/PDGFRβ+ and 36% were CD90+/PDGFRα+.

    “Figure 4a,b shows that within the CD31−/CD45− population, ≈60% were CD90+/PDGFRβ+, and 36% were CD90+/PDGFRα+, respectively.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    VOs contain DLL4+ arterial capillaries, EMCN+ venous capillaries, and α-SMA+ mural cells.

    “Additional immunofluorescence microscopy analyses confirmed the presence of delta‐like protein 4 (DLL4)+ arterial capillaries, endomucin (EMCN)+ veinous capillaries, and α‐smooth muscle actin (α‐SMA)+ mural cells within VOs (Figure 4e–g).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Endothelial capillaries in VSKOs were structurally supported by NG2+ pericytes and contained α-SMA+ mural cells.

    “Further characterization demonstrated that the endothelial capillaries were structurally supported by NG2+ pericytes and contained α‐SMA+ mural cells (Figure 6a,c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Immunofluorescence confirmed presence of a hematopoietic cell lineage in VOs by day 21, with CD34+/CD45+ positive cells next to CD34+ vascular structures.

    “Immunofluorescence microscopy confirmed the presence of a hematopoietic cell lineage in VOs by day 21 of differentiation, where we observed CD34+/CD45+ positive cells next to the CD34+ vascular structures (Figure 8b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CD45+ cells were not visibly present in VOs at day 7, but their numbers significantly increased in subsequent weeks and dispersed throughout the organoids after visible generation of hematopoietic cells on day 21.

    “Although CD45+ cells were not visibly present in VOs at day 7, their numbers significantly increased in the subsequent weeks and dispersed throughout the organoids after the visible generation of hematopoietic cells on day 21 of VO differentiation (Figure 8b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CD68+ cells were present in the dermal region of VSKOs, and almost all CD45+ cells were CD68+.

    “The presence of CD68+ cells in the dermal region of VSKOs was confirmed, where almost all CD45+ cells were CD68+ (Figure 8c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CD45+ cells were homogenously distributed throughout the entire VSKO, predominantly in the dermis with a few cells only in the epidermis.

    “It was observed that the CD45+ cells were homogenously distributed throughout the entire VSKO, predominantly in the dermis with a few cells only in the epidermis.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The vascular-like structures of VSKOs had structural support from NG2+ pericytes and showed α-SMA+ VSMCs.

    “with structural support from NG2+ pericytes, and showed α‐SMA+ VSMCs.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    VSKOs were established from two different hiPSC lines derived from different origins, skin fibroblasts (C32 line) and CD34+ human fetal placental cells (P111 line), demonstrating reproducibility of the differentiation protocol among different hiPSC lines.

    “Here, we established VSKOs from two different hiPSC lines derived from different origins, skin fibroblasts (C32 line) and CD34+ human fetal placental cells (P111 line) and demonstrated the reproducibility of the differentiation protocol among different hiPSC lines.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingnone given

    Adding VEGF to OMM enabled the prolific generation and maintenance of VOs, VSKOs, and sustained VO-derived immune cell differentiation.

    “The conceptualization of adding VEGF to OMM enabled the prolific generation and maintenance of VOs, VSKOs, and sustained VO‐derived immune cell differentiation.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Generating and characterizing VSKOs on an ALI culture system recapitulated vascularized human skin in a physiologically relevant environment, demonstrating macrophages, CD207+ immune cells, neutrophils, and perifollicular vasculature with appropriate luminal structures.

    “Generating and characterizing VSKOs on an ALI culture system recapitulated vascularized human skin in a physiologically relevant environment, which demonstrated macrophages, CD207+ immune cells, neutrophils, and perifollicular vasculature with appropriate luminal structures.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    hiPSC-derived vascular organoids have been developed to provide the full range of cells composing blood vessels.

    “More recently, to provide the full range of cells composing blood vessels, hiPSC‐derived vascular organoids (VO) have been developed.[ 51 ]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [51] R. A. Wimmer, A. Leopoldi, M. Aichinger, D. Kerjaschki, J. M. Penninger, Nat. Protoc. 2019, 14, 85073082. 31554955 source

  • May combine two claims.

    Interpretationown data

    The necessary addition of the GFR-M droplet negatively impacted differentiation of cells derived from the surface ectoderm, affecting hair follicle formation.

    “Consequently, this experiment indicated that the ECFCs were the cause of endothelialization in SKOs (Figure 2b), but the necessary addition of the GFR‐M droplet negatively impacted the differentiation process of cells derived from the surface ectoderm, affecting hair follicle formation (Figure 2c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    VOs undergo heterogeneous differentiation into both arterial and venous lineages and contain mature vascular structures.

    “This data demonstrates a heterogeneous differentiation of VOs into both arterial and venous lineages and highlights the presence of mature vascular structures.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    The VSKO differentiation protocol promotes multi-lineage organoid differentiation and recapitulates the dynamic interplay between vascular and skin tissues.

    “These findings highlight the effectiveness of the VSKO differentiation protocol for promoting multi‐lineage organoid differentiation and recapitulating the dynamic interplay between vascular and skin tissues.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Both P111 and C32 VSKOs developed thick skin layers containing hair follicles, suggesting integration of vascularization and a supportive effect on skin layer formation.

    “In contrast to endothelialized skin organoids using ECFCs, both P111 and C32 hiPSC‐derived VSKOs developed thick skin layers containing hair follicles, suggesting the clear integration of vascularization and their supportive effect on skin layer formation.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    Control groups were established for the co-culture experiments.

    “Control groups were also established (Figure 1b).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationnone given

    The current study developed a VSKO from hiPSCs and showed its superiority to the current SKO and endothelialized SKO models.

    “In the current study, we have developed a VSKO from hiPSCs and showed its superiority to our current SKO and endothelialized SKO models.”
    • Also stated in: § Conclusion · plain
      “To address this limitation, we developed a VSKO from hiPSCs and showed its superiority to our endothelialized SKO model.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Autologous skin graft transplantation is an effective approach for treating major cutaneous wounds.

    “Autologous skin graft transplantation is an effective approach for treating major cutaneous wounds.[ 55 ]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [55] M. L. Ramsey, B. Walker, B. C. Patel, Full‐Thickness Skin Grafts, StatPearls, Treasure Island (FL) 2024. 30422470

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Interpretationown data

    The CD207+ immune cells in VSKOs are either migrating Langerhans cells from the dermis to the epidermis or dermal dendritic cells surrounding nearby hair follicles.

    “demonstrating the presence of either migrating Langerhans cells from the dermis to the epidermis or dermal dendritic cells that were surrounding nearby hair follicles.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Autologous skin grafts are effective in supporting wound closure and reducing contracture.

    “Although autologous skin grafts are effective in supporting wound closure and reducing contracture, their therapeutic benefits are limited by their inconsistency in graft take, lack of donor sites, and high metabolic demands requiring considerable vascular integration.[ 55 ]”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [55] M. L. Ramsey, B. Walker, B. C. Patel, Full‐Thickness Skin Grafts, StatPearls, Treasure Island (FL) 2024. 30422470

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Premisecitation

    Therapeutic benefits of autologous skin grafts are limited by inconsistency in graft take, lack of donor sites, and high metabolic demands requiring considerable vascular integration.

    “Although autologous skin grafts are effective in supporting wound closure and reducing contracture, their therapeutic benefits are limited by their inconsistency in graft take, lack of donor sites, and high metabolic demands requiring considerable vascular integration.[ 55 ]”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [55] M. L. Ramsey, B. Walker, B. C. Patel, Full‐Thickness Skin Grafts, StatPearls, Treasure Island (FL) 2024. 30422470

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Interpretationreasoning

    The SKO + GFP-ECFC co-culture was conducted in a floating culture system, which is suboptimal for skin folliculogenesis.

    “The SKO + GFP‐ECFC co‐culture was conducted in a floating culture system, which is suboptimal for skin folliculogenesis.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisenone given

    The VSKO experiments were maintained in an ALI co-culture system, which is known to promote appendage formation.

    “The VSKO experiments were maintained in an ALI co‐culture system, which is known to promote appendage formation.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    VOs provide not only endothelial cells, like the SKO + GFP-ECFC group, but also perivascular and stromal components that better resemble the dermal niche and may provide essential inductive signals for hair follicle development.

    “In addition, VOs not only provide endothelial cells, like the SKO + GFP‐ECFC group, but also perivascular and stromal components that better resemble the dermal niche and may provide essential inductive signals for hair follicle development.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    The VSKO model enhances the potential applications of vascularized skin constructs for regenerative medicine and opens new avenues for complete in vitro modelling of human skin.

    “Consequently, this model enhances the potential applications of vascularized skin constructs for regenerative medicine applications and opens new avenues for the complete in vitro modelling of human skin.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S2 (checkable). May combine two claims.

    Interpretationreasoning

    The VSKO model demonstrates potential for immunotherapy drug development and disease modelling.

    “In addition, this VSKO model demonstrates potential for immunotherapy drug development and disease modelling, considering drug screening is becoming increasingly popular for new therapeutic strategies, and proves to be a potential future application of the VSKO model.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing).

    Interpretationreasoning

    The recapitulation of hair follicles in the VSKO model can further the potential of current in vitro models.

    “The recapitulation of hair follicles in our VSKO model can further the potential of current in vitro models.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing), S4 (based).

    Interpretationnone given

    The vascularized skin models developed here represent a versatile platform for "clinical trials in a dish" studies.

    “While drug screening studies were not conducted in this study, the vascularized skin models developed here represent a versatile platform for “clinical trials in a dish” studies.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S2 (checkable).

    Interpretationreasoning

    These VSKOs can be used for a range of experimental purposes such as regenerative medicine, drug screening, disease modelling, developmental biology, and biomarker discovery.

    “These VSKOs can be used for a range of experimental purposes such as regenerative medicine, drug screening, disease modelling, developmental biology, and biomarker discovery.”
    Quote in the paperNot a substantial claim
58 of 58

  • The paper relies on this without support.

    Vasculogenesis occurs in human embryos on day 18.

    “Considering vasculogenesis occurs in human embryos on day 18, we recapitulated the process of human skin development and co‐cultured the primed ECFCs with SKOs at day 18 of SKO differentiation (Figure 1a).”

  • The paper relies on this without support.

    Exposing the SKO to air recapitulates the external environment human skin is exposed to.

    “Exposing the SKO to air recapitulates the external environment human skin is exposed to and has shown to enhance differentiation of hiPSC‐derived SKOs.”

  • The paper relies on this without support.

    Application of endothelial cells has limitations including poor integration and limited vascular stability that hinder the formation of functional vasculature in vitro.

    “However, there are identified limitations in the application of endothelial cells, including poor integration, and limited vascular stability which hinder the formation of functional vasculature in vitr o.”

  • The paper relies on this without support.

    Day 115 of differentiation corresponds to approximately 16 weeks of embryonic development.

    “On day 115 of differentiation (≈16‐week of embryonic development)”

  • The paper relies on this without support.

    The immune cells in VSKOs are predominantly myeloid-derived.

    “Considering the immune cells are predominantly myeloid‐derived”
5 of 5

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • hiPSC-derived blood vessel organoids yield self-assembled endothelial networks with lumina and mural support cells capable of forming perfusable vascular structures.

    “In parallel, advances in the generation of hiPSCderived blood vessel organoids (BVOs) have yielded selfassembled endothelial networks with lumina and mural support cells capable of forming perfusable vascular structures.”

    From Vascularized human skin assembloids model compartmental skin organization and immune responsiveness in vitro (2026) · cites this paper as [15]

    The Results section, “Human Vascular Organoids Recapitulate Broad Blood Vessel Phenotypes,” reports hiPSC-derived vascular organoids with endothelial cells, pericytes, mural cells, and capillary structures. The Results section on vascularized skin organoids also describes endothelial capillaries supported by pericytes. Our copy does not establish that the networks have lumina or can form perfusable structures. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • SkOs often display inverted polarity, with the epidermis positioned internally and the dermis externally, and frequently undergo ectopic cartilage differentiation.

    “They often display inverted polarity, in which the epidermis is positioned internally and the dermis externally, and frequently undergo ectopic cartilage differentiation 14,15 .”

    From Vascularized human skin assembloids model compartmental skin organization and immune responsiveness in vitro (2026) · cites this paper as [15]

    The Results section, “Endothelialized Skin Organoids Recapitulate Endothelial Structures in Vitro” (page unavailable), reports abundant cartilage in the SKO + GFP-ECFC group, supporting cartilage formation in that condition. It does not establish that cartilage differentiation is frequent across SKOs. The Results describe epithelial cells shedding into an organoid cyst but do not state that SKOs often have an internal epidermis and external dermis. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

Cites

Flagged first, then by citations

Research works printed but not linked: 12

  1. Yousef H., 2024, Skin (Integument), Epidermis
  2. Kim J. Y., 2025, Physiology, Integument
  3. Hoover E., 2025, Physiology, Hair
  4. Lopez‐Ojeda W., 2025, Skin (Integument)
  5. Brown T. M., 2024, Dermis
  6. Sun J., 2025, Burns Trauma, 13, 070
  7. Hong Z. X., 2023, Mil. Med. Res., 10, 40
  8. Kim K. L., 2009, Biomaterials, 30
  9. Kung E. F., 2008, Dermatol. Surg., 34, 137
  10. Sun X.‐Y., 2022, Elife, 11
  11. Ansari A., 2024, Ectoderm
  12. Tucker W. D., 2024, Blood Vessels

Cited by

Sorted by citations

Related

Authors
Mostina, M., Sun, J., Sim, S. L., Ahmed, I. A., Souza‐Fonesca‐Guimaraes, F., Wolvetang, E. J., Brown, J., Kumari, S., Khosrotehrani, K., & Shafiee, A.
Affiliations
  • The University of Queensland (AU)
  • The University of Queensland Frazer Institute The University of Queensland Woolloongabba Brisbane QLD 4102 Australia
  • Queensland Health (AU)
Corresponding author
  • Khosrotehrani, Kiarash
  • Shafiee, Abbas
ORCID
Venue
Advanced Healthcare Materials
Publisher
Wiley
Published
2025-08-16
Volume
14
Issue
31
Pages
e02108-e02108
ISSN
  • 2192-2640 (ISSN-L)
  • 2192-2659
DOI
10.1002/adhm.202502108
PMC
PMC12683213
OpenAlex
W4413258271
Cited by
17
References
77
Citations by year
2026: 16 · 2025: 1
FWCI
4.28
Type
journal article
Language
en
Open access
Licence
Locations
  • Advanced Healthcare Materials (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Health Sciences › Medicine › Urology › Hair Growth and Disorders
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
Keywords
induced pluripotent stem cells, endothelial colony-forming cells, vascular organoids, skin immune cells, endothelialization, hair follicle morphogenesis, epidermal stratification, skin inflammation
MeSH terms
Cell Differentiation, Humans, Organoids, Skin, Endothelial Cells, Induced Pluripotent Stem Cells

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-23.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-23.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

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