Manufacturing autologous myoblast for regenerative medicine applications.
Cell manufacturing
Myoblast
Myogenic ptosis
Journal
Cytotechnology
ISSN: 0920-9069
Titre abrégé: Cytotechnology
Pays: United States
ID NLM: 8807027
Informations de publication
Date de publication:
Oct 2020
Oct 2020
Historique:
received:
01
05
2020
accepted:
02
09
2020
pubmed:
10
9
2020
medline:
10
9
2020
entrez:
9
9
2020
Statut:
ppublish
Résumé
Autologous myoblasts have been tested in the treatment of muscle-related diseases. However, the standardization of manufacturing myoblasts is still not established. Here we report a flask and animal-free medium-based method of manufacturing clinical-grade myoblast together with establishing releasing criteria for myoblast products under Good Manufacturing Practice (GMP). Quadriceps muscle biopsy samples were donated from three patients with myogenic ptosis. After biopsy samples were digested through enzymatic dissociation, the cells were grown in T175 flasks (passage 0) and hyperflasks (passage 1) in the animal-free SkGM 400 to 500 million myoblast cells were manufactured within 15 to 17 days by the end of passage 1, which met pre-determined releasing criteria. The manufactured myoblast cells could differentiate and fuse into myotubes in vitro, with the possible trend that the donor age may impact the differentiation ability of myoblasts. The present study establishes a flask-based method of manufacturing myoblast in the animal-free medium together with releasing criteria, which is simple, robust, inexpensive and easily reproducible. This study will serve as the validation for a planned phase 1 clinical trial to assess the use of autologous myoblast transplants for the treatment of myogenic ptosis and other myogenic diseases.
Sections du résumé
BACKGROUND
BACKGROUND
Autologous myoblasts have been tested in the treatment of muscle-related diseases. However, the standardization of manufacturing myoblasts is still not established. Here we report a flask and animal-free medium-based method of manufacturing clinical-grade myoblast together with establishing releasing criteria for myoblast products under Good Manufacturing Practice (GMP).
METHODS
METHODS
Quadriceps muscle biopsy samples were donated from three patients with myogenic ptosis. After biopsy samples were digested through enzymatic dissociation, the cells were grown in T175 flasks (passage 0) and hyperflasks (passage 1) in the animal-free SkGM
RESULTS
RESULTS
400 to 500 million myoblast cells were manufactured within 15 to 17 days by the end of passage 1, which met pre-determined releasing criteria. The manufactured myoblast cells could differentiate and fuse into myotubes in vitro, with the possible trend that the donor age may impact the differentiation ability of myoblasts.
CONCLUSIONS
CONCLUSIONS
The present study establishes a flask-based method of manufacturing myoblast in the animal-free medium together with releasing criteria, which is simple, robust, inexpensive and easily reproducible. This study will serve as the validation for a planned phase 1 clinical trial to assess the use of autologous myoblast transplants for the treatment of myogenic ptosis and other myogenic diseases.
Identifiants
pubmed: 32902721
doi: 10.1007/s10616-020-00420-9
pii: 10.1007/s10616-020-00420-9
pmc: PMC7547920
doi:
Types de publication
Journal Article
Langues
eng
Pagination
605-614Références
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