Steroidogenic factor 1 (SF-1) is essential for the development and function of steroidogenic tissues. Stable incorporation of SF-1 into embryonic stem cells has been reported to prime the cells for steroidogenesis. In this study, we established SF1 transgenic mouse embryonic stem cell (SF1-mES cells) and analyzed expression of steroidogenesis-related genes and gonadal lineage-markers. We differentiated mES cells into granulosa-like cells. To test the phenotype for granulosa-like cell, we confirmed transcripts of specific forkhead transcription factor Foxl2 and the follicle stimulating hormone receptor (Fshr). Also, we monitored the expression of EMT-related genes such as E-Cadherin (Cdh1), N-Cadherin (Cdh2), Snai1, Snai2 (Slug), Twist, and Vimentin. We observed the progress into the primitive streakāmesendoderm by gene expression analyses. In addition, the expressions of the steroidogenic enzymes such as 3Ī²-hydroxysteroid dehydrogenase (Hsd3b1), cytochrome P450-containing enzyme (Cyp)-11a1, and Cyp19a1 were time-dependently changed. Next, the mRNA levels of Foxl2 and Fshr representing granulosa-like cell were increased during differentiation of SF1-mES cells. Especially, the level of estradiol and Cdh2 was increased at specific differentiation time. We induced differentiation of mES cells into the functional granulosa-like cells through transfection of mouse SF1 gene. These cells will be useful for further study and potential application of these cells in steroidogenesis