Cell line/type | hESC/hiPSCs-derived CMs (cardiomyocytes) |
---|---|
Species | Human |
Animal free | Yes |
Product | END2-CM |
Xu, X. Q., Graichen, R., Soo, S. Y., Balakrishnan, T., Bte Rahmat, S. N., Sieh, S., ... & Colman, A. (2008). Chemically defined medium supporting cardiomyocyte differentiation of human embryonic stem cells. Differentiation, 76(9), 958-970. In this study, a serum-free medium conditioned by the cell line END2 which efficiently induces hESCs to form cardiomyocytes was analyzed. Insulin, a commonly used medium supplement, acted as a potent inhibitor of cardiomyogenesis in multiple hESC lines and was rapidly cleared by medium conditioning. In the presence of insulin or IGF-1, which also suppressed cardiomyocyte differentiation, the PI3/Akt pathway was activated in undifferentiated hESC, suggesting that insulin/IGF-1 effects were mediated by this signaling cascade. Time course analysis and quantitative RT-PCR revealed impaired expression of endoderm and mesoderm markers in the presence of insulin, particularly if added during early stages of hESC differentiation. Optimized concentrations of PGI2 in a fully synthetic, insulin-free medium resulted in a cardiogenic activity equivalent to END2-CM. Addition of the p38 mitogen-activated protein kinase-inhibitor SB203580, which previously showed to enhance hESC cardiomyogenesis, to these insulin-free and serum-free conditions resulted in a cardiomyocyte content of >10% in differentiated cultures without any preselection. This study represents a significant step toward developing scalable production for cardiomyocytes from hESC using clinically compliant reagents compatible with Good Manufacturing Practice. The formulation of the serum-free medium and the protocol for the development of the conditioned medium is specified in the Materials and Methods, under the heading; Conditioned medium preparation and biochemical analysis. https://www.ncbi.nlm.nih.gov/pubmed/18557764 |
|
Source | Literature - own formulation |
Chemically defined > Yes | Yes |
Contains phenol red > Yes | Yes |
Antibiotics free > Yes | Yes |