CoChat
Sign Up
Cited by 28Extracellular vesicles in diseaseMesenchymal stem cell researchPluripotent Stem Cells Research

A scalable platform for EPSC-induced MSC extracellular vesicles with therapeutic potential

Open access
Gong, Shixin, Li, Nan, Peng, Qinqing, Wang, Feng, Du, Rulong, Zhang, Boyang, Wang, Jian, Han, Le +5 more·2025·Stem Cell Research & Therapy
DOI 10.1186/s13287-025-04507-yPMC PMC12326622
Open this paper in CoChat — free account

Abstract

BACKGROUND: Extracellular Vesicles (EVs) derived from mesenchymal stem cells (MSCs) have gained recognition as promising therapeutic and drug delivery agents in regenerative medicine. However, their clinical application is limited by donor variability, low scalability, and inconsistent therapeutic quality. To overcome these challenges, a robust and standardized production platform is urgently needed. METHODS: We developed a scalable biomanufacturing strategy by generating and expanding MSCs from extended pluripotent stem cells (EPSC) using a suspension bioreactor culture system. A fixed-bed bioreactor was integrated for automated, continuous expansion of iMSCs and downstream EV harvesting. EVs were isolated through a streamlined protocol and characterized for size, morphology, surface markers, and bioactivity. Therapeutic efficacy was assessed in a bleomycin-induced pulmonary fibrosis mouse model. RESULTS: iMSC-derived EVs (iMSC-EVs) exhibited comparable characteristics to primary MSC-EVs, including a size distribution of 70-80 nm, cup-shaped morphology, and expression of canonical EV markers (CD63, CD81, TSG101). iMSCs were expanded for up to 20 days in 3D culture, yielding > 5 × 10⁸ cells per batch using a suspension bioreactor culture system and producing ~ 1.2 × 10¹³ EV particles/day in a fixed-bed bioreactor. In vivo, iMSC-EVs significantly reduced Ashcroft fibrosis scores and bronchoalveolar lavage fluid protein levels in bleomycin-injured lungs, with therapeutic efficacy comparable to primary MSC-EVs. CONCLUSIONS: This study establishes a scalable and standardized platform for producing high-quality iMSC-EVs using bioreactor-based systems. Our approach addresses key limitations in traditional EV production and sets the stage for AI-integrated, fully automated, GMP-compliant manufacturing of therapeutic EVs suitable for clinical translation.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1186/s13287-025-04507-y

Cites

2 of 2 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

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?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 39

  1. [1]Keshtkar S, Azarpira N, Ghahremani MH. Mesenchymal stem cell-derived extracellular vesicles: novel frontiers in regenerative medicine. Stem Cell Res Ther. 2018;9. 10.1186/s13287-018-0791-7. PMC5845209 29523213 · DOI
  2. [2]Weng Z, et al. Therapeutic roles of mesenchymal stem cell-derived extracellular vesicles in cancer. J Hematol Oncol. 2021;14. 10.1186/s13045-021-01141-y. PMC8414028 34479611 · DOI
  3. [3]Tan F, et al. Clinical applications of stem cell-derived exosomes. Signal Transduct Target Ther. 2024;9. 10.1038/s41392-023-01704-0. PMC10784577 38212307 · DOI · Paper page
  4. [4]Wu X, et al. The mechanism of adipose mesenchymal stem cells to stabilize the immune microenvironment of pelvic floor injury by regulating pyroptosis and promoting tissue repair. Mater Today Bio. 2024;24:100910. 10.1016/j.mtbio.2023.100910. 38204481 PMC10776425 · DOI
  5. [5]Kolios G, Paspaliaris V. Mesenchyme stem Cell-Derived conditioned medium as a potential therapeutic tool in idiopathic pulmonary fibrosis. Biomedicines. 2022;10. 10.3390/biomedicines10092298. PMC9496127 36140399 · DOI
  6. [6]Huang R, et al. Plant exosomes fused with engineered mesenchymal stem cell-derived nanovesicles for synergistic therapy of autoimmune skin disorders. J Extracell Vesicles. 2023;12:e12361. 10.1002/jev2.12361. 37859568 PMC10587508 · DOI
  7. [7]Kundu D, Shin SY, Chilian WM, Dong F. The potential of mesenchymal stem Cell-Derived exosomes in cardiac repair. Int J Mol Sci. 2024;25. 10.3390/ijms252413494. PMC11727646 39769256 · DOI
  8. [8]Zhou B, et al. Therapeutic potential of adipose-derived stem cell extracellular vesicles: from inflammation regulation to tissue repair. Stem Cell Res Ther. 2024;15:249. 10.1186/s13287-024-03863-5. 39113098 PMC11304935 · DOI
  9. [9]Monguio-Tortajada M, et al. Nanosized UCMSC-derived extracellular vesicles but not conditioned medium exclusively inhibit the inflammatory response of stimulated T cells: implications for nanomedicine. Theranostics. 2017;7:270–84. 10.7150/thno.16154. 28042333 PMC5197063 · DOI
  10. [10]Abdelgawad M, Bakry NS, Farghali AA, Abdel-Latif A, Lotfy A. Mesenchymal stem cell-based therapy and exosomes in COVID-19: current trends and prospects. Stem Cell Res Ther. 2021;12:469. 10.1186/s13287-021-02542-z. 34419143 PMC8379570 · DOI
  11. [11]Lotfy A, AboQuella NM, Wang H. Mesenchymal stromal/stem cell (MSC)-derived exosomes in clinical trials. Stem Cell Res Ther. 2023;14:66. 10.1186/s13287-023-03287-7. 37024925 PMC10079493 · DOI · Paper page
  12. [12]Monguio-Tortajada M, et al. Acellular cardiac scaffolds enriched with MSC-derived extracellular vesicles limit ventricular remodelling and exert local and systemic Immunomodulation in a myocardial infarction Porcine model. Theranostics. 2022;12:4656–70. 10.7150/thno.72289. 35832072 PMC9254233 · DOI
  13. [13]Monguio-Tortajada M, et al. Local administration of Porcine immunomodulatory, chemotactic and angiogenic extracellular vesicles using engineered cardiac scaffolds for myocardial infarction. Bioact Mater. 2021;6:3314–27. 10.1016/j.bioactmat.2021.02.026. 33778207 PMC7973387 · DOI
  14. [14]Zhang R, et al. An optimized exosome production strategy for enhanced yield while without sacrificing cargo loading efficiency. J Nanobiotechnol. 2022;20:463. 10.1186/s12951-022-01668-3. PMC9618217 36309712 · DOI
  15. [15]Yang YK, Ogando CR, Wang See C, Chang TY, Barabino GA. Changes in phenotype and differentiation potential of human mesenchymal stem cells aging in vitro. Stem Cell Res Ther. 2018;9:131. 10.1186/s13287-018-0876-3. 29751774 PMC5948736 · DOI
  16. [16]Zhou X, et al. The application potential of iMSCs and iMSC-EVs in diseases. Front Bioeng Biotechnol. 2024;12:1434465. 10.3389/fbioe.2024.1434465. 39135947 PMC11317264 · DOI
  17. [17]Giancaterino S, Boi C. Alternative biological sources for extracellular vesicles production and purification strategies for process scale-up. Biotechnol Adv. 2023;63:108092. 10.1016/j.biotechadv.2022.108092. 36608746 · DOI
  18. [18]Costa MHG, et al. Enhanced bioprocess control to advance the manufacture of mesenchymal stromal cell-derived extracellular vesicles in stirred-tank bioreactors. Biotechnol Bioeng. 2023;120:2725–41. 10.1002/bit.28378. 36919232 · DOI
  19. [19]Goral VN, et al. Innovative fixed bed bioreactor platform: enabling linearly scalable adherent cell biomanufacturing with real-time biomass prediction from nutrient consumption. Biotechnol J. 2024;19:e2300635. 10.1002/biot.202300635. 39167554 · DOI
  20. [20]Li R et al. Generation of Blastocyst-like Structures from Mouse Embryonic and Adult Cell Cultures. Cell 179, 687–702 e618. 10.1016/j.cell.2019.09.029 (2019). PMC7359735 31626770 · DOI
  21. [21]Liu B, et al. Chemically defined and xeno-free culture condition for human extended pluripotent stem cells. Nat Commun. 2021;12:3017. 10.1038/s41467-021-23320-8. 34021145 PMC8139978 · DOI
  22. [22]Xu RH, et al. BMP4 initiates human embryonic stem cell differentiation to trophoblast. Nat Biotechnol. 2002;20:1261–4. 10.1038/nbt761. 12426580 · DOI
  23. [23]Wang X, et al. Immune modulatory mesenchymal stem cells derived from human embryonic stem cells through a trophoblast-like stage. Stem Cells. 2016;34:380–91. 10.1002/stem.2242. 26523849 · DOI
  24. [24]Schulz LC, et al. Human embryonic stem cells as models for trophoblast differentiation. Placenta. 2008;29:10–6. 10.1016/j.placenta.2007.10.009. Suppl A. PMC2330081 18054384 · DOI
  25. [25]Koczkowska M, et al. Identifying differentiation markers between dermal fibroblasts and adipose-derived mesenchymal stromal cells (AD-MSCs) in human visceral and subcutaneous tissues using single-cell transcriptomics. Stem Cell Res Ther. 2025;16:64. 10.1186/s13287-025-04185-w. 39934849 PMC11818286 · DOI
  26. [26]Okolicsanyi RK, et al. Human mesenchymal stem cells retain multilineage differentiation capacity including neural marker expression after extended in vitro expansion. PLoS ONE. 2015;10:e0137255. 10.1371/journal.pone.0137255. 26356539 PMC4565666 · DOI
  27. [27]Rao Y, et al. Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses. Stem Cell Res Ther. 2022;13:380. 10.1186/s13287-022-03038-0. 35906661 PMC9338462 · DOI
  28. [28]Kalluri R, LeBleu VS. The biology, function, and biomedical applications of exosomes. Science. 2020;367. 10.1126/science.aau6977. PMC7717626 32029601 · DOI · Paper page
  29. [29]Welsh JA, et al. Minimal information for studies of extracellular vesicles (MISEV2023): from basic to advanced approaches. J Extracell Vesicles. 2024;13:e12404. 10.1002/jev2.12404. 38326288 PMC10850029 · DOI · Paper page
  30. [30]Tsangaris I, Lekka ME, Kitsiouli E, Constantopoulos S, Nakos G. Bronchoalveolar lavage alterations during prolonged ventilation of patients without acute lung injury. Eur Respir J. 2003;21:495–501. 10.1183/09031936.03.00037902. 12662008 · DOI
  31. [31]Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006;126:663–76. 10.1016/j.cell.2006.07.024. 16904174 · DOI · Paper page
  32. [33]Cichocki F, van der Stegen SJC, Miller JS. Engineered and banked iPSCs for advanced NK- and T-cell immunotherapies. Blood. 2023;141:846–55. 10.1182/blood.2022016205. 36327161 PMC10023718 · DOI
  33. [34]Meng W, et al. Engineered mesenchymal stem cell-derived extracellular vesicles constitute a versatile platform for targeted drug delivery. J Control Release. 2023;363:235–52. 10.1016/j.jconrel.2023.09.037. 37739016 · DOI
  34. [36]Batsali AK, et al. Differential expression of cell cycle and WNT pathway-related genes accounts for differences in the growth and differentiation potential of wharton’s jelly and bone marrow-derived mesenchymal stem cells. Stem Cell Res Ther. 2017;8:102. 10.1186/s13287-017-0555-9. 28446235 PMC5406919 · DOI
  35. [37]Lopez-Fernandez A, Garcia-Gragera V, Lecina M, Vives J. Identification of critical process parameters for expansion of clinical grade human wharton’s jelly-derived mesenchymal stromal cells in stirred-tank bioreactors. Biotechnol J. 2024;19:e2300381. 10.1002/biot.202300381. 38403461 · DOI
  36. [38]Dixon AG, Partopour B. Computational fluid dynamics for fixed bed reactor design. Annu Rev Chem Biomol Eng. 2020;11:109–30. 10.1146/annurev-chembioeng-092319-075328. 32151159 · DOI
  37. [39]Klyachko NL, Arzt CJ, Li SM, Gololobova OA, Batrakova EV. Extracellular Vesicle-Based Therapeutics: Preclinical and Clinical Investigations. Pharmaceutics 12. 10.3390/pharmaceutics12121171 (2020). PMC7760239 33271883 · DOI
  38. [40]Fang SB, et al. Small extracellular vesicles derived from human mesenchymal stromal cells prevent group 2 innate lymphoid cell-dominant allergic airway inflammation through delivery of miR-146a-5p. J Extracell Vesicles. 2020;9:1723260. 10.1080/20013078.2020.1723260. 32128074 PMC7034457 · DOI
  39. [41]Haga H, Yan IK, Takahashi K, Matsuda A, Patel T. Extracellular vesicles from bone Marrow-Derived mesenchymal stem cells improve survival from lethal hepatic failure in mice. Stem Cells Transl Med. 2017;6:1262–72. 10.1002/sctm.16-0226. 28213967 PMC5442843 · DOI

Cited by

Sorted by citations

Authors
Gong, S., Li, N., Peng, Q., Wang, F., Du, R., Zhang, B., Wang, J., Han, L., Zhang, Y., Ning, Z., Tan, S., Gu, Y., & Wu, L.
Affiliations
  • Wellcome Sanger Institute (GB)
  • Wellcome/MRC Cambridge Stem Cell Institute (GB)
  • Peking University (CN)
Corresponding author
  • Gong, Shixin
  • Gu, Yuchun
  • Wu, Lida
ORCID
Venue
Stem Cell Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2025-08-05
Volume
16
Issue
1
Pages
426-426
ISSN
1757-6512 (ISSN-L)
DOI
10.1186/s13287-025-04507-y
PMC
PMC12326622
OpenAlex
W4412991157
Cited by
28
References
2
Citations by year
2026: 24 · 2025: 4
FWCI
7.70
Type
journal article
Language
en
Open access
Licence
Locations
  • Stem Cell Research & Therapy (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Apollo (University of Cambridge) (submittedVersion)
Funders
  • Beijing Nova Program (20230484348)
  • National Natural Science Foundation of China (81170236, 31127001)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
Keywords
extracellular vesicles, mesenchymal stem cells, fixed-bed bioreactor, pulmonary fibrosis, bleomycin-induced lung injury, GMP manufacturing, regenerative medicine
MeSH terms
Extracellular Vesicles, Animals, Bleomycin, Humans, Mice, Inbred C57BL, Pulmonary Fibrosis, Cell Culture Techniques, Bioreactors, Pluripotent Stem Cells, Mesenchymal Stem Cell Transplantation, Mice, Mesenchymal 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.

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.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review