Project description:Currently, cardiomyocytes differentiated from human induced pluripotent stem cells (iPSCs) are routinely generated for disease research and drug development as an alternative to animal models. To assess which cell types are present at day 30 of differentiation, we conducted single-cell RNA sequencing using a female and a male line. Our analysis showed that the majority (84-87%) of cells express marker genes typical of cardiomyocytes. Most iPSC-CMs display either ventricular-like (40%) or atrial-like (35-39%) gene expression profiles, whereas a small subset (8-9%) correspond to immature, proliferative cells. The remaining cells expressed extracellular matrix genes and typical epicardial markers. Most likely, these cells underwent suboptimal WNT inhibition following mesoderm induction and differentiated along an epicardial-derived lineage, as reported previously.
Project description:Human induced pluripotent stem cell (IPSC)-derived cardiomyocytes (iCMs) have become important tools to model cardiovascular diseases and drug toxicology. Despite suggested transcriptomic heterogeneity in both iPSC and iCMs, the cellular proteome heterogeneity is poorly understood. Using cutting-edge single cell proteomics, we quantify the maturation from IPSC to iCMs and observed two distinct populations of iCMs with different metabolism, which recapitulates the single adult cardiomyocyte proteome populations albeit less mature.
Project description:Currently, cardiomyocytes differentiated from human induced pluripotent stem cells (iPSCs) are routinely generated for disease research and drug development as an alternative to animal models. Although iPSC-derived cardiomyocytes (iPSC-CMs) are generally assumed to resemble myocytes in the fetal heart, a systematic global comparison is still lacking. We established a robust differentiation protocol to generate mature cardiomyocytes from male and female iPSC lines, and investigated their gene expression and splicing profiles, compared to that of human hearts at different stages of development.
Project description:Dilated cardiomyopathy (DCM) is the leading cause of heart failure and transplantation worldwide. We used iPSCs to model this disease and compared gene expression change before and after gene therapy of cardiomyocytes derived from DCM-specific iPSCs. We used microarrays to detail the global gene expression of patient specific iPSCs, iPSC-derived cardiomyocytes and its response to gene therapy. Skin fibroblasts and iPSCs derived from a family exhibiting familial dilated cardiomyopathy and H7 human ESCs were subjected to RNA extraction and hybridization on Affymetrix microarrays.Global gene expression pattern were compared and analyzed. Cardiomyocytes derived from iPSCs generated from this DCM family were treated with or without adenoriral Serca2a and subjected to RNA extraction and hybridization on Affymetrix microarrays. Global gene expression pattern were compared and analyzed.
Project description:Dilated cardiomyopathy (DCM) is the leading cause of heart failure and transplantation worldwide. We used iPSCs to model this disease and compared gene expression change before and after gene therapy of cardiomyocytes derived from DCM-specific iPSCs. We used microarrays to detail the global gene expression of patient specific iPSCs, iPSC-derived cardiomyocytes and its response to gene therapy.