The Function Study Of VDAC1 In The Induction And Maintenance Of Pluripotency

Embryonic stem cells are cells derived from mammalian early embryos that have the ability to self-renew and differentiate.They can differentiate into any cell type that forms the body and have the ability to develop into a complete individual.Therefore,stem cells have great potential for medical applications,and their basic researches have also received widespread attention.In recent years,the role of metabolic regulation in stem cell fate determination has become increasingly prominent.However,most studies have focused on the regulation of epigenetic modifications by metabolic intermediates,and the research on pluripotency that directly regulates energy metabolism is still relatively rare.It is generally believed that glycolysis dominates energy metabolism in stem cells.In fact,mitochondria are also very important for stem maintenance,but related researches are scarce.In order to explore the relationship between mitochondria and pluripotency,we found through analysis of bioinformatics that a protein VDAC1 on the outer membrane of mitochondria is highly expressed in pluripotent stem cells.As a channel protein,VDAC1 is the pore of mitochondria.It can control various metabolites to enter and exit the mitochondria,and thus regulate cell functions such as energy production and metabolic cross-talk.In addition,VDAC1 is the only channel for ATP to enter and exit the outer membrane of the mitochondria.Producing is crucial.Based on the results of the above investigations,we believe that it is feasible to study VDAC1 on stemness maintenance and regulation.In this experiment,we first explored the expression changes of VDAC1 during MEF reprogramming,and found that as the reprogramming progressed,the expression of VDAC1 gradually increased.Then we utilized CRISPR / Cas9 technology to knock out the VDAC1 gene in mouse embryonic stem cells to construct a VDAC1-deleted ES cell line,and determined that VDAC1-deleted ES cells were still normal diploid karyotypes.Based on this,the energy metabolism of the VDAC1 knockout ES cell line was first detected,and it was found that compared with the wild type,the ES line of the VDAC1 knockout ES line had no significant change in glycolytic capacity,while oxidative phosphorylation capacity obviously decased.Then by examining the ability of cell clone formation and pluripotency gene expression level,it was found that the lack of VDAC1 makes its self-renewal ability significantly lower than that of the wild type.In addition,by comparing the ability of wild-type and VDAC1 knockout cell lines to form embryonic bodies and teratoma,it was found that the absence of VDAC1 decreased the ability of embryonic stem cells to differentiate into the three germ layers.In this study,we successfully constructed mouse embryonic stem cells line in which the mitochondrial channel protein VDAC1 was knocked out,and demonstrated that the absence of VDAC1 reduced the ability to maintain stem cell pluripotency and differentiation,which might related to VDAC1 mediated metabolism.It provides a new model for further exploring the internal mechanisms of energy metabolism and pluripotency.