Effective derivation of ventricular cardiomyocytes from hPSCs using ascorbic acid-containing maturation medium

Kim, Jieun; Kim, Eun‐Mi; Lee, Hyang‐Ae; Kim, Ki‐Suk

doi:10.1080/19768354.2023.2189932

Cited by 2 publications

(2 citation statements)

References 40 publications

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“…However, expression levels of structural proteins (e.g., high α-MHC versus low β-MHC levels) confirmed the expected, immature state of both CDM3 and protein-free-derived CMs. Generally, the expression of cytoskeletal proteins and ion channels was in line with previous results (Additional File 1 , Figure S4) [ 51 , 68 – 70 ].…”

Section: Discussionsupporting

confidence: 91%

Protein-free media for cardiac differentiation of hPSCs in 2000 mL suspension culture

Kriedemann,

Manstein,

Hernandez-Bautista

et al. 2024

Stem Cell Res Ther

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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 and Results 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.

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Section: Discussionsupporting

confidence: 91%

Protein-free media for cardiac differentiation of hPSCs in 2000 mL suspension culture

Kriedemann,

Manstein,

Hernandez-Bautista

et al. 2024

Stem Cell Res Ther

View full text Add to dashboard Cite

show abstract

“…However, expression levels of structural proteins (e.g., high α-MHC versus low β-MHC levels) con rmed the expected, immature state of both CDM3 and protein-free-derived CMs. Generally, the expression of cytoskeletal proteins and ion channels was in line with previous results (Additional File 1, Figure S4) [55,[71][72][73].…”

Section: Discussionsupporting

confidence: 91%

Protein-free media for cardiac differentiation of hPSCs in 2000 mL suspension culture

Kriedemann,

Manstein,

Hernandez-Bautista

et al. 2024

Preprint

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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 & Results Both media compositions enable the efficient, directed differentiation of embryonic and induced hPSCs, matching the cell yields and cardiomyocyte purity ranging from 85-99 % 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.

show abstract

Effective derivation of ventricular cardiomyocytes from hPSCs using ascorbic acid-containing maturation medium

Cited by 2 publications

References 40 publications

Protein-free media for cardiac differentiation of hPSCs in 2000 mL suspension culture

Protein-free media for cardiac differentiation of hPSCs in 2000 mL suspension culture

Protein-free media for cardiac differentiation of hPSCs in 2000 mL suspension culture

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