Engineering 3D micro-compartments for highly efficient and scale-independent expansion of human pluripotent stem cells in bioreactors.
3D culture
Biomicrofluidics
Cell expansion
Human induced pluripotent stem cells
Stem cell niche
Journal
Biomaterials
ISSN: 1878-5905
Titre abrégé: Biomaterials
Pays: Netherlands
ID NLM: 8100316
Informations de publication
Date de publication:
04 2023
04 2023
Historique:
received:
26
03
2022
revised:
12
12
2022
accepted:
31
01
2023
pubmed:
11
2
2023
medline:
11
3
2023
entrez:
10
2
2023
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 36764194
pii: S0142-9612(23)00041-8
doi: 10.1016/j.biomaterials.2023.122033
pii:
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
122033Informations de copyright
Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Feyeux has patent #WO2018096277A1 issued to assignee. Nassoy has patent #WO2018096277A1 issued to assignee. Alessandri has patent #WO2018096277A1 issued to assignee. MF and KA are the founders of TFT; MF, KA, PC and PN are shareholders of Treefrog Therapeutics. MF, KA and PN have a patent pertaining to discoveries presented in this manuscript. Patent no: WO2018096277A1.