Impacts of two types of feeder layer cells ontransformation of bovine iPSCs from primed state to naive state
Feeder cells are essential to derive pluripotent stem cells(PSCs).Mouse embryonic fibro-blasts(MEF)are widely used as feeder to generate and culture embryonic stem cells(ESCs)and induced PSCs(iPSCs)in many species.However,it may not be suitable for livestock ESCs/iPSCs due to interspecies difference.Studies indicate that PSCs can be classified into at least two pluripo-tent states,the naive and the primed states.Previously bovine primed iPSCs were derived from bo-vine Sertoli cells using MEF feeder in our laboratory.Here we compared the effects of MEF feeder and bovine embryonic fibroblasts(BEF)feeder on the maintenance of bovine iPSCs pluripotency and morphology as well their contributions to the naïve-like conversion,based on a naïve medium(NM).The results showed successful conversion of the primed iPSCs to the naive-like iPSCs with-in 3-4 d both on MEF feeder and BEF feeder in NM(termed as MNM and BNM respectively).The colony analysis demonstrated that the colony number,colony alkaline phosphatase,and the conver-sion efficiency were all higher under BNM condition compared with MNM condition.During the long-term cultivation,these bovine iPSCs colonies retained the pluripotent markers of naive iPSCs.Together,bovine naive-like iPSCs can be generated both on MEF and BEF feeder in NM condition.The BNM condition is better to improve the conversion efficiency.
bovinebovine embryonic fibroblastsinduced pluripotent stem cellsmouse embryonic fibroblastsnaïve stateprimed state