Function Research Of Kidins220/ARMS On The Mouse Embryonic Stem Cells

Embryonic stem cells (ESCs), capability to self-renew and differentiate into cells of all three germ layers, are distiguished from othe stem cellsm. ESCs have been broadly used in development research and reproductive medicine.In present study, we screened kidins220/ARMS protein in mouse ESC (mESC) using LC-LTQ assay. RT-PCR analysis suggested that kidins220/ARMS expressed in mESC. This result was confirmed by immunocytochemistry assay using an anti-kidins220/ARMS monoclonal antibody.Kidins220/ARMS knockdown mESCs were established by Jentiviral RNAi system. Kidins220/ARMS knockdown mESCs showed smaller colonies in size. And also the results of growing curve and MTT assay showed us that the proliferation of mESC was remarkably suppressed when Kidins220/ARMS was knock-down. However, no significant difference in clonogenic ability was detected between the knockdown cells and control cells.Alkaline phosphatase (ALP) staining was performed and showed that kidins220/ARMS knockdown did not influence activity of alkaline phosphatase in mESCs. Several pluripotency markers were tested by semi-qRT-PCR, and there was no significant difference in Oct4、Nanog、Rex1、Dnmt31、Dppa4expression between the kidins220/ARMS knockdown cells and control cells. These results suggested that kidins220/ARMS is not necessary for maintaining pluripotency in mESCs. Dppa2and Zfp296expression were upregulated and Salll expression was suppressed, demonstrated by semi-qRT-PCR analysis, implying that kidins220/ARMS may influence the proliferation of mESC via controling Dppa2, Zfp296and Salll expression.Kidins220/ARMS knockdown did not influence embryoid body (EB) formation. However, after adhearing to the gelatin-coated issue culture plate, kidins220/ARMS knockdown EB showed weak adhearing ability, and fewer cells migrated out from EB. Thus kidins220/ARMS may be important for EB further development.In conclusion, we screened kidins220/ARMS protein from mESC by proteomic analysis, and the expression of kidins220/ARMS in mESC was confirmed by RT-PCR and Western blot analysis. Kidins220/ARMS knockdown mESCs were estbilished, and the function of kidins220/ARMS in mESC was tested. Our study provided a new idea and data for understanding the molecular mechanism of self-renew regulation in ESCs.