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

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 thera...

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Main Authors: Shixin Gong, Nan Li, Qinqing Peng, Feng Wang, Rulong Du, Boyang Zhang, Jian Wang, Le Han, Yu Zhang, Zemin Ning, Shengjiang Tan, Yuchun Gu, Lida Wu
Format: Article
Language:English
Published: BMC 2025-08-01
Series:Stem Cell Research & Therapy
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Online Access:https://doi.org/10.1186/s13287-025-04507-y
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author Shixin Gong
Nan Li
Qinqing Peng
Feng Wang
Rulong Du
Boyang Zhang
Jian Wang
Le Han
Yu Zhang
Zemin Ning
Shengjiang Tan
Yuchun Gu
Lida Wu
author_facet Shixin Gong
Nan Li
Qinqing Peng
Feng Wang
Rulong Du
Boyang Zhang
Jian Wang
Le Han
Yu Zhang
Zemin Ning
Shengjiang Tan
Yuchun Gu
Lida Wu
author_sort Shixin Gong
collection DOAJ
description 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.
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spelling doaj-art-02871e6a7f764451bd0f18dd681e6a772025-08-20T03:04:30ZengBMCStem Cell Research & Therapy1757-65122025-08-0116111910.1186/s13287-025-04507-yA scalable platform for EPSC-Induced MSC extracellular vesicles with therapeutic potentialShixin Gong0Nan Li1Qinqing Peng2Feng Wang3Rulong Du4Boyang Zhang5Jian Wang6Le Han7Yu Zhang8Zemin Ning9Shengjiang Tan10Yuchun Gu11Lida Wu12Allife Medicine Co., LtdAllife Medicine Co., LtdAllife Medicine Co., LtdAllife Medicine Co., LtdAllife Medicine Co., LtdAllife Medicine Co., LtdAllife Medicine Co., LtdAllife Medicine Co., LtdAllife Medicine Co., LtdThe Wellcome Sanger InstituteCambridge Institute for Medical ResearchAllife Medicine Co., LtdAllife Medicine Co., LtdAbstract 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.https://doi.org/10.1186/s13287-025-04507-yExtracellular vesiclesExtended pluripotent stem cellsInduced mesenchymal stem cellsScalable biomanufacturingPulmonary fibrosis therapyRegenerative medicine
spellingShingle Shixin Gong
Nan Li
Qinqing Peng
Feng Wang
Rulong Du
Boyang Zhang
Jian Wang
Le Han
Yu Zhang
Zemin Ning
Shengjiang Tan
Yuchun Gu
Lida Wu
A scalable platform for EPSC-Induced MSC extracellular vesicles with therapeutic potential
Stem Cell Research & Therapy
Extracellular vesicles
Extended pluripotent stem cells
Induced mesenchymal stem cells
Scalable biomanufacturing
Pulmonary fibrosis therapy
Regenerative medicine
title A scalable platform for EPSC-Induced MSC extracellular vesicles with therapeutic potential
title_full A scalable platform for EPSC-Induced MSC extracellular vesicles with therapeutic potential
title_fullStr A scalable platform for EPSC-Induced MSC extracellular vesicles with therapeutic potential
title_full_unstemmed A scalable platform for EPSC-Induced MSC extracellular vesicles with therapeutic potential
title_short A scalable platform for EPSC-Induced MSC extracellular vesicles with therapeutic potential
title_sort scalable platform for epsc induced msc extracellular vesicles with therapeutic potential
topic Extracellular vesicles
Extended pluripotent stem cells
Induced mesenchymal stem cells
Scalable biomanufacturing
Pulmonary fibrosis therapy
Regenerative medicine
url https://doi.org/10.1186/s13287-025-04507-y
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