Objective:In this study,human embryonic stem cells(hESCs) were induced to differentiate into hypothalamic neural progenitor cells in vitro,and the effects of different concentrations of androgen on the differentiation of hypothalamic neural progenitor cells were compared. Methods:The quality of human embryonic stem cell line CCRM22 was identified,CCRM22 was induced to differentiate into hypothalamic neural progenitor cells in vitro,then 1×10-8 mol/L and 1×10-7 mol/L testosterone were added to the differentiation medium(anhydrous ethanol as cosolvent control). The differentiated cells were collected at different stages of differentiation,and the differentiation efficiency was compared. The differentiated cells were identified by RT-qPCR,flow cytometry and immnofluorescence,and the expression of genes related to reproductive function regulation was detected. Results:The differentiation efficiency of CCRM22 into hypothalamic neural progenitor cells was more than 85%,but testosterone treatment decreased its differentiation efficiency;differentiated cells expressed nerve cell marker NESTIN and hypothalamic neural progenitor cell specific marker NKX2.1,expressed both KISS1 and androgen receptor(AR),and the expression of AR was positively correlated with testosterone concentration. Conclusion:Human embryonic stem cells were successfully induced into hypothalamic neural progenitor cells. High concentration of testosterone inhibited the differentiation of hESCs into hypothalamic neural progenitor cells. This cell model can be used to study the effects of hyperandrogenic environment in early pregnancy on hypothalamic nerve cell differentiation and development of offspring in women with polycystic ovary syndrome in vitro.