Facile bead-to-bead cell-transfer method for serial subculture and large-scale expansion of human mesenchymal stem cells in bioreactors.
bead-to-bead transfer
bioreactor
mesenchymal stem cell
microcarrier
nucleosides
scale up
Journal
Stem cells translational medicine
ISSN: 2157-6580
Titre abrégé: Stem Cells Transl Med
Pays: England
ID NLM: 101578022
Informations de publication
Date de publication:
09 2021
09 2021
Historique:
revised:
29
03
2021
received:
10
11
2020
accepted:
21
04
2021
pubmed:
20
5
2021
medline:
7
4
2022
entrez:
19
5
2021
Statut:
ppublish
Résumé
The conventional planar culture of adherent cells is inefficient for large-scale manufacturing of cell and gene therapy products. We developed a facile and efficient bead-to-bead cell-transfer method for serial subculture and large-scale expansion of human mesenchymal stem cells (hMSCs) with microcarriers in bioreactors. We first compared culture medium with and without nucleosides and found the former maintained the expression of surface markers of hMSCs during their prolonged culture and enabled faster cell proliferation. Subsequently, we developed our bead-to-bead cell transfer method to subculture hMSCs and found that intermittent agitation after adding fresh microcarriers to cell-populated microcarriers could promote spontaneous cell migration to fresh microcarriers, reduce microcarrier aggregation, and improve cell yield. This method enabled serial subculture of hMSCs in spinner flasks from passage 4 to passage 9 without using proteolytic enzymes, which showed faster cell proliferation than the serial planar cultures undergoing multiple enzyme treatment. Finally, we used the medium containing nucleosides and our bead-to-bead cell transfer method for cell culture scale-up from 4- to 50-L cultures in single-use bioreactors. We achieved a 242-fold increase in the number of cells to 1.45 × 10
Identifiants
pubmed: 34008349
doi: 10.1002/sctm.20-0501
pmc: PMC8380445
doi:
Substances chimiques
Culture Media
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1329-1342Informations de copyright
© 2021 Showa Denko Materials Co., Ltd. STEM CELLS TRANSLATIONAL MEDICINE published by Wiley Periodicals LLC on behalf of AlphaMed Press.
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