Sweat Gland Organoids Originating from Reprogrammed Epidermal Keratinocytes Functionally Recapitulated Damaged Skin

Signaling pathways involved in regulation of stem cell self-renewal, including MAPK, JAK-STAT, Hippo, and FoxO pathways, were upregulated in ISO-treated HEKs.

“some important signaling pathways involved in regulation of stem cell self‐renewal, including MAPK,[ 32 , 33 ] JAK‐STAT,[ 34 ] Hippo,[ 35 ] and FoxO[ 36 ] pathways, were upregulated in ISO‐treated HEKs (Figure S5b, Supporting Information).”

• cites [32] Q.‐L. Ying, J. Wray, J. Nichols, L. Batlle‐Morera, B. Doble, J. Woodgett, P. Cohen, A. Smith, Nature 2008, 453, 519. 18497825 PMC5328678 sourceSupported
• cites [33] H. Chen, R. Guo, Q. Zhang, H. Guo, M. Yang, Z. Wu, S. Gao, L. Liu, L. Chen, Proc. Natl. Acad. Sci. U. S. A. 2015, 112, E5936. 26483458 PMC4640739 sourceSupported
• cites [34] R. R. Stine, E. L. Matunis, Adv. Exp. Med. Biol. 2013, 786, 247. 23696361 sourceNot checked — no full text of the source
• cites [35] B. Zhao, K. Tumaneng, K.‐L. Guan, Nat. Cell Biol. 2011, 13, 877. 21808241 PMC3987945 sourceSupported

SwGs are maintained by stem and progenitor cells within the ducts and coiled glands.

“SwGs are maintained by stem and progenitor cells within the ducts and coiled glands.[ 2 ]”

• cites [2] C. P. Lu, L. Polak, A. S. Rocha, H. A. Pasolli, S.‐C. Chen, N. Sharma, C. Blanpain, E. Fuchs, Cell 2012, 150, 136. 22770217 PMC3423199 sourcePartly supported

  The Discussion section “Four Populations of Stem Cells in Adult Pawskin” reports homeostatic turnover in sweat ducts and identifies progenitors in the coiled glands. It also says the glands show little, if any, turnover during normal homeostasis; glandular progenitors proliferated in response to targeted injury. Thus, the source does not establish that progenitors in both compartments actively maintain sweat glands under normal conditions. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

EDA signaling has a key role in SwG initiation and development.

“Ectodysplasin‐A (EDA) signaling has a key role in SwG initiation and development.[ 13 , 14 ]”

• cites [13] C.‐Y. Cui, M. Kunisada, D. Esibizione, E. G. Douglass, D. Schlessinger, J. Invest. Dermatol. 2009, 129, 984. 18923450 PMC5155335 sourceSupported
• cites [14] C.‐Y. Cui, M. Yin, J. Sima, V. Childress, M. Michel, Y. Piao, D. Schlessinger, Development 2014, 141, 3752. 25249463 PMC4197578 sourceSupported

A previous study demonstrated the conversion of bone marrow-derived mesenchymal stem cells into SwG cells via forced EDA gene expression.

“Our previous study demonstrated the conversion of bone marrow‐derived mesenchymal stem cells into SwG cells via forced EDA gene expression,[ 4 ]”

• cites [4] S. Sun, J. Xiao, J. Huo, Z. Geng, K. Ma, X. Sun, X. Fu, Stem Cell Res. Ther. 2018, 9, 8. 29329593 PMC5766979 sourceSupported

Several studies have suggested an unanticipated level of cellular plasticity within epidermal keratinocytes, resulting in cell-fate flexibility during wound repair.

“Several studies have suggested an unanticipated level of cellular plasticity within epidermal keratinocytes, resulting in cell‐fate flexibility during wound repair.[ 8 , 9 , 10 , 11 , 12 ]”

• cites [8] X. Sun, X. Fu, W. Han, M. Zhao, L. Chalmers, Expert Opin. Biol. Ther. 2013, 13, 901. 23517553 sourceNot checked — no full text of the source
• cites [9] C. Blanpain, E. Fuchs, Science 2014, 344, 1242281. 24926024 PMC4523269 sourceSupported
• cites [10] A. Trond, Nat. Biotechnol. 2008, 26, 1276. 18931654Not matched

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

• cites [11] N. Maherali, T. Ahfeldt, A. Rigamonti, J. Utikal, C. Cowan, K. Hochedlinger, Cell Stem Cell 2008, 3, 340. 18786420 PMC3987901 sourcePartly supported

  The Discussion reports that neonatal foreskin keratinocytes were experimentally reprogrammed to a pluripotent state, supporting cellular fate flexibility under induced conditions. It does not address keratinocyte fate changes during wound repair, so it cannot support that setting-specific part of the claim. Our copy provides section labels but no page numbers. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [12] K. A. U. Gonzales, E. Fuchs, Dev. Cell 2017, 43, 387. 29161590 PMC5797699 sourceSupported

Organoids developed in artificial in vitro conditions retain the organ identity.

“Organoids developed in the artificial in vitro conditions retain the organ identity,[ 16 , 17 ]”

• cites [16] A. Fatehullah, S. H. Tan, N. Barker, Nat. Cell Biol. 2016, 18, 246. 26911908 sourceSupported
• cites [17] E. R. Shamir, A. J. Ewald, Nat. Rev. Mol. Cell Biol. 2014, 15, 647. 25237826 PMC4352326 sourceSupported

The formation of organoids recapitulates a 3D in vivo-like microenvironment that may compensate for the lack of structural support for tissue formation in the damaged extracellular matrix environment when used as xenografts.

“the formation of organoids also recapitulate a 3D in vivo‐like microenvironment that may well compensate for the lack of structural support for tissue formation in the damaged extracellular matrix environment when used as xenografts.[ 18 , 19 ]”

• cites [18] S. Sugimoto, Y. Ohta, M. Fujii, M. Matano, M. Shimokawa, K. Nanki, S. Date, S. Nishikori, Y. Nakazato, T. Nakamura, T. Kanai, T. Sato, Cell Stem Cell 2018, 22, 171. 29290616 sourceNot checked — no full text of the source
• cites [19] H. Hu, H. Gehart, B. Artegiani, C. Löpez‐Iglesias, F. Dekkers, O. Basak, J. Van Es, S. M. Chuva De Sousa Lopes, H. Begthel, J. Korving, M. Van Den Born, C. Zou, C. Quirk, L. Chiriboga, C. M. Rice, S. Ma, A. Rios, P. J. Peters, Y. P. De Jong, H. Clevers, Cell 2018, 175, 1591. 30500538 sourcePartly supported

  The Summary (p. 3) describes a 3D organoid culture that retains key hepatocyte features, and the engraftment results (p. 13) show that its cells repopulate damaged mouse livers. The paper does not say that organoids compensate for deficient structural support in damaged extracellular matrix. Indeed, the engraftment methods (pp. 12, 24) say the organoids were digested into single cells before transplantation, rather than used as intact 3D xenografts. (LLM, conf=0.89)

  Experimental — accuracy not yet measured

Human SwG organoids derived from SwG epithelial cells contributed to regeneration of epidermis and SwGs in vivo.

“Human SwG organoids derived from SwG epithelial cells contributed to the regeneration of epidermis and SwGs in vivo.[ 22 ]”

• cites [22] J. Diao, Cell Death Dis. 2019, 10, 238. 30858357 PMC6411741 sourcePartly supported

  The Results section reports that transplanted sweat-gland-organoid-derived cells participated in epidermal and sweat gland regeneration in vivo. However, the organoids were generated from mouse sweat gland epithelial cells and transplanted into mice; the paper does not report this result for human organoids. (LLM, conf=0.92)

  Experimental — accuracy not yet measured

PCA and hierarchical clustering revealed that iSwGCs generated from the reprogramming procedure were clustered with hSwGCs but separated from untreated HEKs and HEK-EDA cells without SGM treatment in the PC1 dimension.

“principle component analysis (PCA) and hierarchical clustering analysis revealed that iSwGCs generated from our reprogramming procedure were clustered with hSwGCs but separated from the untreated HEKs and HEK‐EDA cells without SGM treatment in the PC1 dimension (Figure 3d,e)”

Long-term exposure to SGM upregulated CK18, AQP5 and α-SMA transcription in HEK-EDA cells.

“although long‐term exposure to SGM upregulated CK18, aquaporin 5 (AQP5) and α‐SMA transcription in HEK‐EDA cells (Figure S2c, Supporting Information)”

Functional sweating can be induced in mouse paws upon β2-AR activation by intradermal injection of 5 μg/mL isoproterenol.

“functional sweating could be induced in mouse paws upon β 2‐AR activation by intradermal injection of 5 µg mL−1 isoproterenol (ISO, Figure S4c, Supporting Information).”

ISO stimulation enriched genes related to gland morphogenesis and development in HEKs.

“ISO stimulation enriched genes related to gland morphogenesis and development in HEKs (Figure S7b, Supporting Information).”

After 18 days' culture, iSwGOs maintained in microfluidic chambers formed typical tubular structures with hollow lumens lined with a simple epithelium.

“After 18 days’ culture, the iSwGOs maintained in microfluidic chambers formed typical tubular structures with hollow lumens that were lined with a simple epithelium (Figure 4l and Figure S14d, Supporting Information).”

RNA-seq showed that iSwGCs expressed major SwG genes, channel genes, and carbohydrate metabolic genes.

“the iSwGCs expressed major SwG genes, channel genes, and carbohydrate metabolic genes (Figure S11, Supporting Information)”

iSwGOs could expand to 200 μm in diameter within 4 weeks.

“These iSwGC‐derived organoids (iSwGOs) could expand to 200 µm in diameter within 4 weeks.”

HEKs are efficiently converted into SwG cells (iSwGCs) by forced expression of EDA combined with specific SwG culture medium.

“by forced expression of the ectodysplasin‐A in human epidermal keratinocytes (HEKs) combined with specific SwG culture medium, HEKs are efficiently converted into SwG cells (iSwGCs).”

EDA overexpression in LGR5hi HEKs conferred significantly higher levels of BMP5, CEA, CK18, CK19, CK5, α-SMA, and AQP5 expression when cultured in SGM compared with LGR5lo HEKs.

“EDA overexpression in LGR5hi cells conferred significantly higher levels of BMP5, CEA, CK18, CK19, CK5, α‐SMA, and AQP5 expression when cells were cultured in SGM (Figure S3b,c, Supporting Information).”

iSwGOs elongated with a bipolar shape along the bFGF gradient direction 8 days after exposure to bFGF gradients.

“the elongation of iSwGOs with a bipolar shape along the gradient direction was observed 8 days after exposure to bFGF gradients (Figure S14c, Supporting Information).”

β2-AR is strongly expressed in CK18+/CK19+ luminal glandular cells in adult SwGs.

“β 2‐AR was strongly expressed in the CK18+/CK19+ luminal glandular cells in adult SwGs (Figure 1c and Figure S4a,b, Supporting Information).”

Tet-on CRISPR conditional EDA activation in HaCaT keratinocytes reprogrammed them into SwG-like cells after 2 days' exposure to SGM supplemented with ISO.

“After 2 days’ exposure to SGM supplemented with ISO, the expression of dCas9‐E and EDA targeting guides reprogrammed HaCaT cells into SwG‐like cells (Figures S9c–f and S10a–d, Supporting Information).”

β2-AR inhibition by ICI-118551 or β2-AR siRNAs attenuated the effect of ISO on upregulating stemness markers in HEKs.

“Importantly, β 2‐AR inhibition by ICI‐118551 (Figure 1g,i,j) or β 2‐AR siRNAs (Figure 1h,k,l) attenuated the effect of ISO, which further demonstrates the importance of β 2‐ARs in upregulating the stemness of HEKs.”

Stimulating β2-AR with ISO significantly increased the stemness of HEKs.

“these data showed that stimulating β 2‐AR with ISO significantly increased the stemness of HEKs”

iSwGCs were similar in morphology to native SwG cells.

“Given that the iSwGCs were similar in morphology to native SwG cells (Figure 3a )”

Genes critical to gland development, actin cytoskeleton organization and regulation, and cell secretion were upregulated in iSwGCs.

“genes critical to gland development, actin cytoskeleton organization and regulation, and cell secretion were also upregulated in iSwGCs (Figure S11, Supporting Information)”

BMP5, EDA, EDAR, FGF18, WNT5A ligand and its downstream target ID1 were expressed at relatively higher levels in ISO-treated HEKs compared with controls.

“Of these, BMP5, EDA, EDAR, FGF18, WNT5A ligand and its downstream target ID1 were expressed at relatively higher levels as compared to controls (Figure S7c, Supporting Information).”

Single iSwGCs embedded in matrigel and cultured in SGM induction medium initially formed a solid ball and self-organized into monoclonal spheres within 2 weeks.

“single iSwGCs were embedded in matrigel and cultured in SGM induction medium. Under this situation, single cells initially formed a solid ball and self‐organized into monoclonal spheres within 2 weeks (Figure 4a ).”

Spherical organoids derived from HEKs gradually progressed into a lumen-containing morphology with visible lumen surrounded by single- or multiple-layered epithelial cells after 2 to 3 weeks' culture in SGM supplemented with ISO.

“the spherical organoids derived from HEKs gradually progressed into a lumen‐containing morphology with visible lumen surrounded by single‐ or multiple‐layered epithelial cells after 2 to 3 weeks’ culture in SGM supplemented with ISO (Figure 4b–e).”

Only a small proportion of reprogrammed HEK-EDA cells expressed ductal (CK5+/CK10+, 5.01±1.47%), luminal (CK18+/AQP5+, 6.31±1.64%) or myoepithelial (CK5+/α-SMA+, 3.37±0.87%) markers.

“only a small proportion of the reprogrammed cells could be induced to express the ductal (CK5+/CK10+, 5.01±1.47%), luminal (CK18+/AQP5+, 6.31±1.64%) or myoepithelial (CK5+/α‐SMA+, 3.37±0.87%) markers”

Intracellular Ca2+ activity was present in the epithelial cells of iSwGOs after exposure to a Ca2+ fluorescent indicator.

“we observed the presence of intracellular Ca2+ activity in the epithelial cells of iSwGOs after exposure to a Ca2+ fluorescent indicator (Figure 4k).”

After 8 days of induction, qRT-PCR revealed a significant increase in levels of CK18, AQP5, and α-SMA but not CK5 in iSwGCs compared with primary isolated HEKs.

“After 8 days of induction, RNA expression profiling by qRT‐PCR revealed a significant increase in levels of key SwG genes including CK18, AQP5, and α‐SMA but not CK5 in iSwGCs as compared with primary isolated HEKs (Figure 2c).”

Inhibition of β2-AR with ICI-118551 reversed the ISO-induced BMP5, EDA, CK5, CK18, CK19, CEA expression.

“inhibition of β 2‐AR with ICI‐118551 reversed the ISO‐induced BMP5, EDA, CK5, CK18, CK19, CEA expression (Figure S7e,f, Supporting Information)”

A panel of stemness-related genes including LGR5, OCT4, ESRG, MYC, KLF2, and FOXQ1 were significantly enriched in HEKs after ISO stimulation (p < 0.05, FDR ≤ 0.05, |FC| ≥ 2).

“a panel of stemness‐related genes including LGR5, OCT4, ESRG, MYC, KLF2, and FOXQ1were significantly enriched (p < 0.05, [FDR] ≤ 0.05, and |FC| ≥ 2) in HEKs after ISO stimulation (Figure 1d–f).”

Organoids with distinctive layers could be consistently passaged and stably maintained under the 3D culture condition for more than 3 months.

“The organoids with distinctive layers could be consistently passaged and stably maintained under the 3D culture condition for more than 3 months.”

HEKs were converted into iSwGCs by combining β2-AR stimulation, forced transgenic expression of EDA, and SGM culture.

“we designed the strategy in Figure 2a to systemically reprogram HEKs into iSwGCs by combining stimulation with β 2‐AR, forced transgenic expression of EDA, and SGM culture.”

iSwGCs showed the gene expression patterns of functional SwGs for a set of genes involved in iron transport, protein transport, carbohydrate metabolism, and glycoprotein metabolism.

“iSwGCs showed the gene expression patterns of functional SwGs for a set of genes involved in iron transport, protein transport, carbohydrate metabolism, and glycoprotein metabolism (Figure 3f).”

ISO added in the medium could support the growth of large organoids in iSwGCs.

“Also, ISO added in the medium could support the growth of large organoids in iSwGCs (Figure S12a,b, Supporting Information).”

ISO treatment of HEKs caused significant changes in RNA-seq results, identifying 5314 known DEGs, 2823 upregulated and 2491 downregulated (FDR ≤ 0.05 and |FC| ≥ 2.0).

“we used the β 2‐AR agonist ISO to stimulate HEKs and found significant changes in RNA‐seq results, identifying a total of 5314 known DEGs, 2823 upregulated and 2491 downregulated, in ISO‐treated HEKs (Figure S5a, Supporting Information, false discovery rate [FDR] ≤ 0.05 and fold change |FC| ≥ 2.0).”

iSwGCs had a similar level of cytoplasmic free Ca2+ intensity as hSwGCs.

“we measured the intracellular Ca2+ signals by using an orange fluorescent calcium indicator and found a similar level of cytoplasmic free Ca2+ intensity in iSwGCs as in hSwGCs (Figure 3c).”

Upregulation of some specific SwG markers was observed in ISO-treated HEKs.

“Upregulation of some specific SwG markers was also observed in ISO‐treated HEKs (Figure S7e,f, Supporting Information).”

Hair follicle-related genes were downregulated in iSwGCs.

“In contrast, hair follicle‐related genes were downregulated (Figure 2c), which indicated that most HEK‐converted cells engaged a SwG fate.”

ISO treatment upregulated LGR5, LGR6, OCT4, and SOX9 expression in HEKs.

“Consistently, ISO treatment upregulated LGR5, LGR6, OCT4, and SOX9 expression in HEKs (Figure 1g–l).”

After single-cell plating, the 3D culture system mimicking the paracrine environment of in vivo SwGs led to rapid expansion of the engineered organoids.

“After single‐cell plating, the 3D culture system mimicking the paracrine environment of in vivo SwGs led to rapid expansion of the engineered organoids.”

• AssumptionRests on an assumption: The 3D culture system mimics the paracrine environment of in vivo SwGs.

Nascent ductal and glandular structures were seen in the dermis of recipient mice at 21 days after transplantation.

“nascent ductal and glandular structures were seen in the dermis of recipient mice at 21 days after transplantation (Figure 5d).”

iSwGOs formed typical tubular structures with hollow lumens lined with a simple epithelium.

“formed typical tubular structures with hollow lumens that were lined with a simple epithelium (Figure 4l and Figure S14d, Supporting Information).”

Rete ridges in iSwGO-engrafted paw skin were elongated and intertwined with underlying dermal tissues comparable to those derived from normal skin.

“Note that the rete ridges in iSwGO‐engrafted paw skin were elongated and intertwined with underlying dermal tissues comparable to those derived from normal skin.”

EDA-GFP+ cells were seen in newly generated glandular structures.

“EDA‐GFP+ cells were seen in the newly generated glandular structures.”

Systemic transcriptome profiling and function enrichment analysis showed that gene expression patterns of related GO terms in iSwGCs were similar to those of native SwG cells.

“Systemic transcriptome profiling and function enrichment analysis showed consistently that the gene expression patterns of related GO terms in iSwGCs were similar to those of native SwG cells.”

The number of mice positive for starch-iodine reaction increased time-dependently.

“The number of mice positive for starch–iodine reaction increased time‐dependently”

iSwGOs were able to self-assemble into tubular structures along a bFGF gradient in vitro.

“An important feature is that the iSwGOs were able to self‐assemble into tubular structures along a bFGF gradient in vitro.”

On starch-iodine sweat tests, only paws of iSwGO-treated mice showed indigo-black dots at day 21 after iSwGO transplantation.

“Strikingly, on starch–iodine sweat tests, only paws of iSwGO‐treated mice showed indigo‐black dots at day 21 after iSwGO transplantation (Figure 5b)”

Immunostaining of human-specific histone protein confirmed the human origin of GFP+ glandular structures in the mouse paw skin.

“Of note, immunostaining of human‐specific histone protein further confirmed the human origin of GFP+ glandular structures in the mouse paw skin (Figure 5f).”

GFP-labeled cells were robustly observed in the regenerative epidermis and nascent glands.

“GFP‐labeled cells were robustly observed in the regenerative epidermis and nascent glands.”

The presence of BMP4 and CHIR99021 improved the efficiency for the formation of CK5+/α-SMA+ iSwGOs during 3D cultures.

“The presence of BMP4 and CHIR99021 improved the efficiency for the formation of CK5+/α‐SMA+ iSwGOs during 3D cultures.”

Increasing the stemness of HEKs could promote the reprogramming efficiency.

“This finding suggested that increasing the stemness of HEKs could promote the reprogramming efficiency.”

Activation of β2-AR signaling plays a role in priming HEKs for the SwG fate.

“so activation of β 2‐AR signaling plays a role on priming HEKs for the SwG fate.”

The resulting iSwGOs retained the typical cyto-organization and histological features closely resembling the native SwG bulk in vivo.

“The resulting iSwGOs retained the typical cyto‐organization and histological features closely resembling the native SwG bulk in vivo.”

The two-step strategy efficiently reprogrammed HEKs into SwG cells.

“Taken together, these data demonstrate that we developed an efficient two‐step strategy to reprogram HEKs into SwG cells.”

Most HEK-converted cells engaged a SwG fate.

“which indicated that most HEK‐converted cells engaged a SwG fate.”

iSwGCs could develop into iSwGOs via long-term 3D culture.

“Collectively, these results demonstrate that iSwGCs could develop into iSwGOs via long‐term 3D culture.”

β2-ARs are important in upregulating the stemness of HEKs.

“which further demonstrates the importance of β 2‐ARs in upregulating the stemness of HEKs.”

Stimulating β2-AR with ISO might have the potential to promote the efficiency of cell-fate conversion to SwG cells via EDA reprogramming.

“and might have the potential to promote the efficiency of the cell‐fate conversion to SwG cells via EDA reprogramming.”

EDA alone was not sufficient to reprogram HEKs into SwG cells.

“Therefore, EDA alone was not sufficient to reprogram HEKs into SwG cells.”

The indigo-black dots reflect functional restoration of SwGs in thermally injured mice.

“reflecting a functional restoration of SwGs in the thermally injured mice.”

iSwGOs were capable of responding to the chemotaxis of extracellular signaling molecules to form tubular structures in vitro.

“which suggests that the iSwGOs were capable of responding to the chemotaxis of extracellular signaling molecules to form tubular structures in vitro.”

iSwGOs derived from reprogrammed HEKs successfully repopulated damaged skin with fully restored SwGs in vivo, structurally and functionally.

“Collectively, our data demonstrate that iSwGOs derived from reprogrammed HEKs successfully repopulated damage skin with fully restored SwGs in vivo.”

• Also stated in: § Discussion · plain

  “Another key feature of the iSwGOs is their ability to repopulate damaged skin structurally and functionally.”

Deep partial-thickness scald burns in foot pads of mice produced deep dermal injury with complete destruction of SwGs.

“we created deep partial‐thickness scald burns in foot pads of mice, which produced a deep dermal injury with complete destruction of the SwGs.”

EDA can act as an intrinsic factor to re-establish a SwG cell identity during SwG reprogramming.

“which suggests that EDA can act as an intrinsic factor to re‐establish a SwG cell identity during SwG reprogramming.”

EDA expression primed an intrinsic possibility for driving the fate conversion of HEKs.

“EDA expression primed an intrinsic possibility for driving the fate conversion of HEKs”

iSwGCs could self-assemble into organoids in a special 3D culture system.

“Taken together, these results demonstrate that iSwGCs could self‐assemble into organoids in a special 3D culture system”

In our study, we established functional human SwG organoids from iSwGCs.

“In our study, we established functional human SwG organoids from iSwGCs.”

We demonstrate a practical and feasible strategy for achieving functional SwG regeneration by using engineering human SwG organoids in vivo.

“In conclusion, we demonstrate a practical and feasible strategy for achieving functional SwG regeneration by using engineering human SwG organoids in vivo.”

• AssumptionRests on an assumption: Lack of donor tissues and low ex vivo expansion capacity of primary SwG cells hamper transplantation of autologous or xenogenous SwG-derived cells for therapy in the clinic.
• AssumptionRests on an assumption: Wound-resident SwG progenitor cells are not sufficient to orchestrate the healing process for promotion of de novo SwG morphogenesis.
• AssumptionRests on an assumption: In patients with large skin defects, the pathological microenvironment within the wound bed has been a major roadblock for functional reconstruction of damaged skin.

Activation of β2-AR signaling efficiently improved the SwG cell conversion from stably transduced human keratinocytes by priming cellular stemness and gland development-associated gene expression programs within the initial HEKs.

“Specifically, activation of β 2‐AR signaling efficiently improved the SwG cell conversion from stably transduced human keratinocytes by priming cellular stemness and gland development‐associated gene expression programs within the initial HEKs.”

Regeneration of functional SwG organoids from reprogrammed HEKs highlights great translational potential for personalized SwG regeneration in patients with large skin defects.

“Regeneration of functional SwG organoids from reprogrammed HEKs highlights the great translational potential for personalized SwG regeneration in patients with large skin defects.”

Spatio-temporal biological cues from the extracellular microenvironment may play crucial roles in guiding and extending the morphogenetic process of SwGs in vitro and in vivo.

“so spatio‐temporal biological cues from the extracellular microenvironment may play crucial roles in guiding and extending the morphogenetic process of SwGs in vitro and in vivo.”

Transplantation of SwG organoids for regeneration is highly feasible because their initial size is close to that of native sweat buds during SwG development.

“for SwG organoids, the initial size is close to that of native sweat buds during SwG development, so their transplantation for regeneration is highly feasible.”

• AssumptionRests on an assumption: The initial size of SwG organoids is close to that of native sweat buds during SwG development.

Generation of iSwGCs highly depended on the addition of SwG conditional medium.

“whereas generation of iSwGCs highly depended on the addition of SwG conditional medium.”

With this strategy, it is possible to generate human SwG organoids at a sufficiently large scale for clinical use.

“With this strategy, it is possible to generate human SwG organoids at a sufficiently large scale for clinical use.”

De novo SwG regeneration rarely occurs during wound healing.

“As a result, de novo SwG regeneration rarely occurs during wound healing.”

Reprogrammed SwG cells can be generated from HEKs transduced with a single factor, EDA, under defined SwG induction conditions.

“we first showed that reprogrammed SwG cells can be generated from HEKs transduced with a single factor, EDA, under defined SwG induction conditions.”

iSwGO engraftment promoted wound closure in vivo.

“iSwGO engraftment also promoted wound closure in vivo.”

iSwGOs were able to respond to extracellular biological cues for further development toward functional SwGs.

“and were able to respond to the extracellular biological cues for further development toward functional SwGs.”