Matrix-free human pluripotent stem cell manufacturing by seed train approach and intermediate cryopreservation.
Aggregate dissociation
GMP
Intermediate cryopreservation
STBR
Seed train
Suspension culture
hPSC
Journal
Stem cell research & therapy
ISSN: 1757-6512
Titre abrégé: Stem Cell Res Ther
Pays: England
ID NLM: 101527581
Informations de publication
Date de publication:
25 Mar 2024
25 Mar 2024
Historique:
received:
10
01
2024
accepted:
17
03
2024
medline:
26
3
2024
pubmed:
26
3
2024
entrez:
26
3
2024
Statut:
epublish
Résumé
Human pluripotent stem cells (hPSCs) have an enormous therapeutic potential, but large quantities of cells will need to be supplied by reliable, economically viable production processes. The suspension culture (three-dimensional; 3D) of hPSCs in stirred tank bioreactors (STBRs) has enormous potential for fuelling these cell demands. In this study, the efficient long-term matrix-free suspension culture of hPSC aggregates is shown. STBR-controlled, chemical aggregate dissociation and optimized passage duration of 3 or 4 days promotes exponential hPSC proliferation, process efficiency and upscaling by a seed train approach. Intermediate high-density cryopreservation of suspension-derived hPSCs followed by direct STBR inoculation enabled complete omission of matrix-dependent 2D (two-dimensional) culture. Optimized 3D cultivation over 8 passages (32 days) cumulatively yielded ≈4.7 × 10 Together, an entirely matrix-free, highly efficient, flexible and automation-friendly hPSC expansion strategy is demonstrated, facilitating the development of good manufacturing practice-compliant closed-system manufacturing in large scale.
Sections du résumé
BACKGROUND
BACKGROUND
Human pluripotent stem cells (hPSCs) have an enormous therapeutic potential, but large quantities of cells will need to be supplied by reliable, economically viable production processes. The suspension culture (three-dimensional; 3D) of hPSCs in stirred tank bioreactors (STBRs) has enormous potential for fuelling these cell demands. In this study, the efficient long-term matrix-free suspension culture of hPSC aggregates is shown.
METHODS AND RESULTS
RESULTS
STBR-controlled, chemical aggregate dissociation and optimized passage duration of 3 or 4 days promotes exponential hPSC proliferation, process efficiency and upscaling by a seed train approach. Intermediate high-density cryopreservation of suspension-derived hPSCs followed by direct STBR inoculation enabled complete omission of matrix-dependent 2D (two-dimensional) culture. Optimized 3D cultivation over 8 passages (32 days) cumulatively yielded ≈4.7 × 10
CONCLUSIONS
CONCLUSIONS
Together, an entirely matrix-free, highly efficient, flexible and automation-friendly hPSC expansion strategy is demonstrated, facilitating the development of good manufacturing practice-compliant closed-system manufacturing in large scale.
Identifiants
pubmed: 38528578
doi: 10.1186/s13287-024-03699-z
pii: 10.1186/s13287-024-03699-z
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
89Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : EXC 62/2
Organisme : Deutsche Forschungsgemeinschaft
ID : ZW64/4-2
Organisme : Deutsche Forschungsgemeinschaft
ID : KFO311 / ZW64/7-1
Organisme : Bundesministerium für Bildung und Forschung
ID : 01EK1601A
Organisme : Bundesministerium für Bildung und Forschung
ID : 13XP5092B
Organisme : Bundesministerium für Bildung und Forschung
ID : 031L0249
Organisme : Bundesministerium für Bildung und Forschung
ID : 01EK2108A
Organisme : Förderung aus Mitteln des Niedersächsischen Vorab
ID : ZN3340
Organisme : HORIZON EUROPE Health
ID : 101056712
Informations de copyright
© 2024. The Author(s).
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