Bmi-1, a polycomb gene family member, plays an important role in cell cycle regulation, cell immortalization, and cell senescence. Recently, numerous studies have demonstrated that Bmi-1 is involved in the regulation of self-renewal and differentiation of stem cells. However, the molecular mechanism underlying this biological process remains largely unclear. In the present review, we summarized the function of Bmi-1 as a transcriptional regulator of gene expression, with particular reference to stem cells.