Abstract: Growth of buffalo preantral follicles in vitro culture was studied to investigate the effects of size of preantral follicles, methods of culture, relative factors. In experiment 1, Effects of ascorbic acid (Vc), FSH and EGF on the in vitro development of buffalo preantral follicles were examined. Ascorbic acid can maintain the follicular morphologic integrity during culture but does not affect the survival and growth of follicles. FSH can promote the growth of follicles, but does not affect the survival and morphology of follicles. The growth promotive role of FSH is enhanced by the presence of Vc. EGF can accelerate the diameter increasing of follicles in the early culture stage, and this action is enhanced in the presence of FSH. In experiment 2,The in vitro growth of follicles in different diameters (60~180µm) was examined. As the follicular diameter increases, the survival rate and growing extent of follicles increases, the morphological abnormity of follicles decreases. In experiment 2, Conventional, two dimension (agarose coating) and three dimension (agarose embed) culture system were compared in their efficiency of supporting follicle growth. Three dimensions culture system is proved to be suitable for culturing the follicles in diameter of 60~100µm, while the two dimension culture system is proved to be suitable for culturing the follicles in diameter of 100~140µm.

Key words: buffalo , preantral follicle, in vitro culture