Development of in vitro technologies that will allow the culture of early stage follicles before antral stage is an essential part of research in reproductive biology in order to understand the ovarian folliculogenesis better. Current evidence suggests that oocyte and somatic cells-derived growth factors interacting with each other and extracellular matrix proteins at paracrine level are involved in this early, gonadotrophin-independent phase of follicle growth. Basement membrane matrix protein (Matrigel™) is a soluble gel rich in extracellular matrix proteins and growth factors. Activin A promotes preantral follicle growth in vivo by inducing the proliferation of granulosa cells and by upregulating the expression of FSH receptor and aromatase enzyme. We hypothesized that activin A and matrigel may provide a better in vitro environment for early stage preantral follicles. Preantral follicles isolated from 14-21 day old BALB/c mice were cultured in matrigel ± activin A for four days. The growth (119.4% versus 45.4%, p < 0.05; respectively) and survival rates (76.3% versus 43.7%, p < 0.05; respectively) of the follicles treated with activin A were significantly higher compared to those without activin A. These results suggest that Activin A and matrigel provide a better in vitro milieu for the growth of isolated ovarian follicles.