Cyclosporin A Promotes Invasion And Migration Of Extravillous Trophoblast Cells Derived From Human Embryonic Stem Cells And Human Induced Pluripotent Stem Cells

Background Extravillous trophoblast(EVT)cells play an essential role in the maternal-fetal interaction.Althoughabnormal development and function of EVT cells,including impaired migration and invasion capability,are believed to be etiologically linked to severe pregnancy disorders including pre-eclampsia(PE),the associated molecular mechanisms are unclear due to the lack of an appropriate cell model in vitro.Cyclosporine A(CsA)is a macrolide immunosuppressant and is also used in clinic to improve pregnancy outcomes.However,whether CsA has any effects on the function of EVT cells has not been well investigated.Objective In this study,we induced differentiation of human embryonic stem cells(hESCs)and human induced pluripotent stem cells(hiPSCs)into EVT cells(hESCs-EVT cells and hiPSCs-EVT cells,respectively).We then investigated the effects of CsA on the invasion and migration of these derived EVT cells.Methods HESCs and hiPSCs were cultured in a culture medium containing NRG1,A83-01,Y27632 and matrigel,and we collected these derived EVT cells by flow cytometry for sorting cells positive for double HLA-G and KRT7,whichare EVT markers.We then investigated the effects of CsA on the invasion and migration of these derived EVT cells.Results We found that the hiPSCs and hESCs changed from a colony-like morphology to an epithelial-like morphology at day 3,and began to form cyst-like structures at day 6.We then found that the hESCs-EVT and hiPSCs-EVT cells expressed highlevels of the EVT markers suchas KRT7,ITGA5 and HLA-G but low levels of OCT4,a stem cell marker.Finally,we found CsA significantly promoted the invasion and migration of hESCs-EVT and hiPSCs-EVT cells.Conclusions In summary,we successfully generated hiPSC/hESC-derived human EVT cells,whichmay be applicable for investigating the remodeling process of spiral arteries remodeling and the possible mechanisms of EVT-related diseases in vitro.Furthermore,our findings provide direct evidence that CsA regulates the function of EVT cells and molecular basis by whichCsA may be used to treat pregnancy complications in clinic associated withdeficient EVT function.