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Engineering 3D micro-compartments for highly efficient and scale-independent expansion of human pluripotent stem cells in bioreactors

Philippe J. R. Cohen, Elisa Luquet, Justine Pletenka, Andrea Leonard, Elise Warter, Basile Gurchenkov, Jessica Carrere, Clement Rieu, Jerome Hardouin, Fabien Moncaubeig, et al.
(2023) BIOMATERIALS. 295.
Author
Organization
Abstract
Human pluripotent stem cells (hPSCs) have emerged as the most promising cellular source for cell therapies. To overcome the scale-up limitations of classical 2D culture systems, suspension cultures have been developed to meet the need for large-scale culture in regenerative medicine. Despite constant improvements, current protocols that use microcarriers or generate cell aggregates only achieve moderate amplification performance. Here, guided by reports showing that hPSCs can self-organize in vitro into cysts reminiscent of the epiblast stage in embryo development, we developed a physio-mimetic approach for hPSC culture. We engineered stem cell niche microenvironments inside microfluidics-assisted core-shell microcapsules. We demonstrate that lumenized three-dimensional colonies significantly improve viability and expansion rates while maintaining pluripotency compared to standard hPSC culture platforms such as 2D cultures, microcarriers, and aggregates. By further tuning capsule size and culture conditions, we scale up this method to industrial-scale stirred tank bioreactors and achieve an unprecedented hPSC amplification rate of 277-fold in 6.5 days. In brief, our findings indicate that our 3D culture system offers a suitable strategy both for basic stem cell biology experiments and for clinical applications.
Keywords
Human induced pluripotent stem cells, 3D culture, Cell expansion, Stem cell niche, Biomicrofluidics, DIFFERENTIATION, CULTURE, PROLIFERATION, MORPHOGENESIS, QUALITY, OXYGEN, CYCLE

Citation

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MLA
Cohen, Philippe J. R., et al. “Engineering 3D Micro-Compartments for Highly Efficient and Scale-Independent Expansion of Human Pluripotent Stem Cells in Bioreactors.” BIOMATERIALS, vol. 295, 2023, doi:10.1016/j.biomaterials.2023.122033.
APA
Cohen, P. J. R., Luquet, E., Pletenka, J., Leonard, A., Warter, E., Gurchenkov, B., … Alessandri, K. (2023). Engineering 3D micro-compartments for highly efficient and scale-independent expansion of human pluripotent stem cells in bioreactors. BIOMATERIALS, 295. https://doi.org/10.1016/j.biomaterials.2023.122033
Chicago author-date
Cohen, Philippe J. R., Elisa Luquet, Justine Pletenka, Andrea Leonard, Elise Warter, Basile Gurchenkov, Jessica Carrere, et al. 2023. “Engineering 3D Micro-Compartments for Highly Efficient and Scale-Independent Expansion of Human Pluripotent Stem Cells in Bioreactors.” BIOMATERIALS 295. https://doi.org/10.1016/j.biomaterials.2023.122033.
Chicago author-date (all authors)
Cohen, Philippe J. R., Elisa Luquet, Justine Pletenka, Andrea Leonard, Elise Warter, Basile Gurchenkov, Jessica Carrere, Clement Rieu, Jerome Hardouin, Fabien Moncaubeig, Michael Lanero, Eddy Quelennec, Helene Wurtz, Emilie Jamet, Maelle Demarco, Celine Banal, Paul Van Liedekerke, Pierre Nassoy, Maxime Feyeux, Nathalie Lefort, and Kevin Alessandri. 2023. “Engineering 3D Micro-Compartments for Highly Efficient and Scale-Independent Expansion of Human Pluripotent Stem Cells in Bioreactors.” BIOMATERIALS 295. doi:10.1016/j.biomaterials.2023.122033.
Vancouver
1.
Cohen PJR, Luquet E, Pletenka J, Leonard A, Warter E, Gurchenkov B, et al. Engineering 3D micro-compartments for highly efficient and scale-independent expansion of human pluripotent stem cells in bioreactors. BIOMATERIALS. 2023;295.
IEEE
[1]
P. J. R. Cohen et al., “Engineering 3D micro-compartments for highly efficient and scale-independent expansion of human pluripotent stem cells in bioreactors,” BIOMATERIALS, vol. 295, 2023.
@article{01HQ136CHSDSV1MYJ8M5SBNEHM,
  abstract     = {{Human pluripotent stem cells (hPSCs) have emerged as the most promising cellular source for cell therapies. To overcome the scale-up limitations of classical 2D culture systems, suspension cultures have been developed to meet the need for large-scale culture in regenerative medicine. Despite constant improvements, current protocols that use microcarriers or generate cell aggregates only achieve moderate amplification performance. Here, guided by reports showing that hPSCs can self-organize in vitro into cysts reminiscent of the epiblast stage in embryo development, we developed a physio-mimetic approach for hPSC culture. We engineered stem cell niche microenvironments inside microfluidics-assisted core-shell microcapsules. We demonstrate that lumenized three-dimensional colonies significantly improve viability and expansion rates while maintaining pluripotency compared to standard hPSC culture platforms such as 2D cultures, microcarriers, and aggregates. By further tuning capsule size and culture conditions, we scale up this method to industrial-scale stirred tank bioreactors and achieve an unprecedented hPSC amplification rate of 277-fold in 6.5 days. In brief, our findings indicate that our 3D culture system offers a suitable strategy both for basic stem cell biology experiments and for clinical applications.}},
  articleno    = {{122033}},
  author       = {{Cohen, Philippe J. R. and  Luquet, Elisa and  Pletenka, Justine and  Leonard, Andrea and  Warter, Elise and  Gurchenkov, Basile and  Carrere, Jessica and  Rieu, Clement and  Hardouin, Jerome and  Moncaubeig, Fabien and  Lanero, Michael and  Quelennec, Eddy and  Wurtz, Helene and  Jamet, Emilie and  Demarco, Maelle and  Banal, Celine and Van Liedekerke, Paul and  Nassoy, Pierre and  Feyeux, Maxime and  Lefort, Nathalie and  Alessandri, Kevin}},
  issn         = {{0142-9612}},
  journal      = {{BIOMATERIALS}},
  keywords     = {{Human induced pluripotent stem cells,3D culture,Cell expansion,Stem cell niche,Biomicrofluidics,DIFFERENTIATION,CULTURE,PROLIFERATION,MORPHOGENESIS,QUALITY,OXYGEN,CYCLE}},
  language     = {{eng}},
  pages        = {{13}},
  title        = {{Engineering 3D micro-compartments for highly efficient and scale-independent expansion of human pluripotent stem cells in bioreactors}},
  url          = {{http://doi.org/10.1016/j.biomaterials.2023.122033}},
  volume       = {{295}},
  year         = {{2023}},
}
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