Alternating 2D and 3D culture reduces cell size and extends the lifespan of placenta-derived mesenchymal stem cells
BackgroundMesenchymal stem cells (MSCs) hold great promise for treating a variety of human diseases; however, their clinical translation is hindered by challenges in large‐scale expansion while preserving therapeutic potency and maintaining small cell size. Conventional 2D culture on rigid substrate...
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Frontiers Media S.A.
2025-08-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2025.1632810/full |
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| author | Ying Pan Li Han Li Han Yakun Yang Xinran Wu Aijun Wang Aijun Wang Aijun Wang Liangqi Xie Wuqiang Zhu Shue Wang Yuguo Lei Yuguo Lei |
| author_facet | Ying Pan Li Han Li Han Yakun Yang Xinran Wu Aijun Wang Aijun Wang Aijun Wang Liangqi Xie Wuqiang Zhu Shue Wang Yuguo Lei Yuguo Lei |
| author_sort | Ying Pan |
| collection | DOAJ |
| description | BackgroundMesenchymal stem cells (MSCs) hold great promise for treating a variety of human diseases; however, their clinical translation is hindered by challenges in large‐scale expansion while preserving therapeutic potency and maintaining small cell size. Conventional 2D culture on rigid substrates induces MSC senescence and enlargement, compromising their function and biodistribution.MethodsWe present an alternating 2D/3D culture strategy that combines adherent monolayer expansion with transient spheroid formation to mitigate these limitations. Placenta‐derived MSCs were cultured under optimized spheroid conditions, with extracellular matrix supplementation and chemically defined media to enhance viability. To address scalability, we developed RGD-functionalized alginate hydrogel tubes (AlgTubes) that enable dynamic transitions between adherent and spheroid states for continuous culture.ResultsSpheroid culture significantly reduced cell size and enhanced immunomodulatory function. The alternating 2D/3D protocol slowed MSC enlargement and senescence over multiple passages while preserving anti-inflammatory activity. Extracellular matrix supplementation and chemically defined media further improved cell viability. AlgTubes successfully supported the alternating culture strategy in a continuous and scalable format.Conclusions The alternating 2D/3D culture system effectively overcomes limitations of conventional MSC expansion by mitigating enlargement, delaying senescence, and preserving both proliferative capacity and immunoregulatory potency. Combined with AlgTube technology, this work demonstrates a promising strategy for MSC manufacturing |
| format | Article |
| id | doaj-art-10aee8dcb7f64bcda646294b5ea4f88e |
| institution | Kabale University |
| issn | 2296-4185 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj-art-10aee8dcb7f64bcda646294b5ea4f88e2025-08-22T05:26:49ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852025-08-011310.3389/fbioe.2025.16328101632810Alternating 2D and 3D culture reduces cell size and extends the lifespan of placenta-derived mesenchymal stem cellsYing Pan0Li Han1Li Han2Yakun Yang3Xinran Wu4Aijun Wang5Aijun Wang6Aijun Wang7Liangqi Xie8Wuqiang Zhu9Shue Wang10Yuguo Lei11Yuguo Lei12Department of Biomedical Engineering, Pennsylvania State University, University Park, PA, United StatesDepartment of Biomedical Engineering, Pennsylvania State University, University Park, PA, United StatesHuck Institutes of Life Sciences, Pennsylvania State University, University Park, PA, United StatesDepartment of Biomedical Engineering, Pennsylvania State University, University Park, PA, United StatesDepartment of Biomedical Engineering, Pennsylvania State University, University Park, PA, United StatesDepartment of Biomedical Engineering, University of California, Davis, CA, United StatesDepartment of Surgery, Center for Surgical Bioengineering, School of Medicine, University of California, Davis, Sacramento, CA, United StatesInstitute for Pediatric Regenerative Medicine, Shriners Hospitals for Children, Sacramento, CA, United StatesCancer Biology and Infection Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, United StatesDepartment of Cardiovascular Medicine, Physiology and Biomedical Engineering, Center for Regenerative Medicine, Mayo Clinic Arizona, Scottsdale, AZ, United StatesDepartment of Chemistry, Chemical and Biomedical Engineering, University of New Haven, West Haven, CT, United StatesDepartment of Biomedical Engineering, Pennsylvania State University, University Park, PA, United StatesHuck Institutes of Life Sciences, Pennsylvania State University, University Park, PA, United StatesBackgroundMesenchymal stem cells (MSCs) hold great promise for treating a variety of human diseases; however, their clinical translation is hindered by challenges in large‐scale expansion while preserving therapeutic potency and maintaining small cell size. Conventional 2D culture on rigid substrates induces MSC senescence and enlargement, compromising their function and biodistribution.MethodsWe present an alternating 2D/3D culture strategy that combines adherent monolayer expansion with transient spheroid formation to mitigate these limitations. Placenta‐derived MSCs were cultured under optimized spheroid conditions, with extracellular matrix supplementation and chemically defined media to enhance viability. To address scalability, we developed RGD-functionalized alginate hydrogel tubes (AlgTubes) that enable dynamic transitions between adherent and spheroid states for continuous culture.ResultsSpheroid culture significantly reduced cell size and enhanced immunomodulatory function. The alternating 2D/3D protocol slowed MSC enlargement and senescence over multiple passages while preserving anti-inflammatory activity. Extracellular matrix supplementation and chemically defined media further improved cell viability. AlgTubes successfully supported the alternating culture strategy in a continuous and scalable format.Conclusions The alternating 2D/3D culture system effectively overcomes limitations of conventional MSC expansion by mitigating enlargement, delaying senescence, and preserving both proliferative capacity and immunoregulatory potency. Combined with AlgTube technology, this work demonstrates a promising strategy for MSC manufacturinghttps://www.frontiersin.org/articles/10.3389/fbioe.2025.1632810/fullmesenchymal stem cellsalternating 2D/3D culturespheroid culturecell sizesenescence |
| spellingShingle | Ying Pan Li Han Li Han Yakun Yang Xinran Wu Aijun Wang Aijun Wang Aijun Wang Liangqi Xie Wuqiang Zhu Shue Wang Yuguo Lei Yuguo Lei Alternating 2D and 3D culture reduces cell size and extends the lifespan of placenta-derived mesenchymal stem cells Frontiers in Bioengineering and Biotechnology mesenchymal stem cells alternating 2D/3D culture spheroid culture cell size senescence |
| title | Alternating 2D and 3D culture reduces cell size and extends the lifespan of placenta-derived mesenchymal stem cells |
| title_full | Alternating 2D and 3D culture reduces cell size and extends the lifespan of placenta-derived mesenchymal stem cells |
| title_fullStr | Alternating 2D and 3D culture reduces cell size and extends the lifespan of placenta-derived mesenchymal stem cells |
| title_full_unstemmed | Alternating 2D and 3D culture reduces cell size and extends the lifespan of placenta-derived mesenchymal stem cells |
| title_short | Alternating 2D and 3D culture reduces cell size and extends the lifespan of placenta-derived mesenchymal stem cells |
| title_sort | alternating 2d and 3d culture reduces cell size and extends the lifespan of placenta derived mesenchymal stem cells |
| topic | mesenchymal stem cells alternating 2D/3D culture spheroid culture cell size senescence |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2025.1632810/full |
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