Rapamycin Promotes Cardiomyocyte Differentiation of Human Induced Pluripotent Stem Cells in a Stage-Dependent Manner.

Stem Cells and Development
Min JiangFeng Cao

Abstract

Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) are a promising source for cardiac regenerative therapy, and ideal for in vitro cell modeling of cardiovascular diseases and drug screening. Recent studies have shown that rapamycin can promote cardiomyocyte differentiation in various stem cells. However, how rapamycin affects cardiomyocyte differentiation of iPSCs is still not fully understood. This study aimed to investigate the effect of rapamycin on cardiomyocyte differentiation based on embryoid body (EB) method. First, to determine the autophagy induction protocol, different concentrations of rapamycin were applied in hEBs on day 6. The autophagy was most significant when applying rapamycin at 1 μM for 48 h, demonstrating by the LC3II/LC3I ratio and p62 expression. Then, 1 μM rapamycin was applied for 48 h at different time points of cardiomyocyte differentiation to investigate the role of rapamycin in this process. Compared with control, rapamycin applied on days 0-4 of differentiation significantly decreased the proportion of beating EBs and expression of cardiomyocyte-specific genes, while rapamycin applied on days 4-14 significantly increased them. Among all groups, rapamycin applied on days 4-6 achieved ...Continue Reading

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Methods Mentioned

BETA
Protein Assay
Fluorescence
FACS
transmission electron microscopy
flow cytometry

Software Mentioned

Image J
FACS Diva
SPSS

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