Protein-free media for cardiac differentiation of hPSCs in 2000 mL suspension culture.
Stem Cell Res Ther
; 15(1): 213, 2024 Jul 18.
Article
en En
| MEDLINE
| ID: mdl-39020441
ABSTRACT
BACKGROUND:
Commonly used media for the differentiation of human pluripotent stem cells into cardiomyocytes (hPSC-CMs) contain high concentrations of proteins, in particular albumin, which is prone to quality variations and presents a substantial cost factor, hampering the clinical translation of in vitro-generated cardiomyocytes for heart repair. To overcome these limitations, we have developed chemically defined, entirely protein-free media based on RPMI, supplemented with L-ascorbic acid 2-phosphate (AA-2P) and either the non-ionic surfactant Pluronic F-68 or a specific polyvinyl alcohol (PVA). METHODS ANDRESULTS:
Both media compositions enable the efficient, directed differentiation of embryonic and induced hPSCs, matching the cell yields and cardiomyocyte purity ranging from 85 to 99% achieved with the widely used protein-based CDM3 medium. The protein-free differentiation approach was readily up-scaled to a 2000 mL process scale in a fully controlled stirred tank bioreactor in suspension culture, producing > 1.3 × 109 cardiomyocytes in a single process run. Transcriptome analysis, flow cytometry, electrophysiology, and contractile force measurements revealed that the mass-produced cardiomyocytes differentiated in protein-free medium exhibit the expected ventricular-like properties equivalent to the well-established characteristics of CDM3-control cells.CONCLUSIONS:
This study promotes the robustness and upscaling of the cardiomyogenic differentiation process, substantially reduces media costs, and provides an important step toward the clinical translation of hPSC-CMs for heart regeneration.Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Diferenciación Celular
/
Medios de Cultivo
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Miocitos Cardíacos
Límite:
Humans
Idioma:
En
Año:
2024
Tipo del documento:
Article