Research Of C-jun In Maintenance And Escape Of Pluripotency In Human Embryonic Stem Cells

Pluripotent stem cells(PSCs)have the potential to self-renew and differentiate into three germ layers.They are ideal cell models for the study of cell fate regulation mechanism and have broad application prospects in regenerative therapy.However,there are still many problems in clinical application.The main reasons include that the mechanism of pluripotent stem cells in the process of fate determination is not thoroughly studied,and the regulation mechanism of pluripotent maintenance and withdrawal is still not clear.Previous studies in our lab showed that c-Jun significantly affected the maintenance and exit of pluripotency of mouse embryonic stem cells by regulating the Ss18 forms phase transition and other pathways,and at the same time,as a "parallel mirror factor" of the core pluripotency gene Oct4,it antagonized the gain of pluripotency,and proposed the interface hypothesis of somatic cells and pluripotent stem cells.However,the role of c-Jun in the fate determination of human embryonic stem cells is still poorly understood,and the similarities and differences between human and mouse systems remain unclear.In this study,CRISPR/Cas9 technology was used to successfully construct c-Jun knockout human embryonic stem cell lines.In combination with morphological characteristics,detection of pluripotent gene expression and teratoma formation,it was found that c-Jun knockout had no significant effect on pluripotency maintenance and the early stage of three germ layers differentiation of human embryonic stem cells.To further determine the functional regulatory role of c-Jun in pluripotent withdrawal and the differentiation into the terminal functional lineage of the late three germ layers of human embryonic stem cells,in this study,human embryonic stem cells were directed to differentiate into cardiomyocytes,liver cells,kidney cells,nerve cells,somatic precursor cells,and definite endoderm cells,it was found that c-Jun knockout significantly inhibited the differentiation of human embryonic stem cells into cardiomyocytes,but promoted the differentiation of human embryonic stem cells into hepatocytes,which was different from the phenotype of c-Jun knockout mice that had been reported to inhibit the differentiation of hepatocytes but had no effect on the differentiation of cardiomyocytes.In addition,c-Jun knockout had no significant effect on the differentiation of human embryonic stem cells into kidney cells,nerve cells,somatic precursor cells,and definite endoderm cells which is consistent with the phenotype that c-Jun knockout mice had no effect on the development and differentiation of these cells.The role of c-Jun in pluripotency maintenance and escape of human embryonic stem cells in this study was preliminary studied,which laid a theoretical foundation for further exploration of the fate determination of pluripotent stem cells and comparative study of the mechanism of c-Jun in the fate regulation of functional cell of the lineages in different species.It has important theoretical significance for the safe and effective application of pluripotent stem cells in clinical research and disease treatment in the future.