Study of Cardiac Organoids Model Construction Based on Human Induced Pluripotent Stem Cells
In this study,human induced pluripotent stem cells(hiPSCs)were used to construct an in vitro organoid-based substitution model for drug cardiotoxicity.The hiPSCs were cultured to determine the optimal differentiation generation,and pluripotency was identified using immunofluorescence staining.The hiPSCs were seeded into ultra-low adsorption 96-well plates at different cell densities,and the medium containing growth factors and signaling pathway inhibitors were added for inducing differentiation,and the morphological changes and pulsatile characteristics were recorded by light microscopy.Cardiac organoids at different time points in the differentiation process were collected,and real-time reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was adopted to detect the relative expression of cell-specific genes of cardiac organoids.The results showed that the resuscitated hiPSCs grew well with clear edges and excellent cell pluripotency.The morphology of the differentiated cardiac organoids at a density of 5 000 cells per well was the best,and trended to be a relatively stable state for 8-10 d culturing,and a relatively stable state could be maintained within 18 d.RT-qPCR results showed that specific genes of cardiomyocytes and other cardiac components,such as endothelial cells,smooth muscle cells,and fibroblasts,were expressed.The establishment of cardiac organoid models in this study can more realistically and accurately simulate the biological characteristics and functions of the heart in vivo,which lays a foundation for the subsequent use of cardiac organoid models for potential cardiotoxicity studies of drugs in the early stage of research and development.
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