关键词: 水牛; 卵巢颗粒细胞; 原代培养; 培养基; 增殖; 凋亡

Abstract: The purpose of this study is to compare the growth of buffalo ovarian granulosa cells (GCs) cultured with different culture medium, and research the proliferation, karyotype and apoptosis of GCs to know well the growth characteristic of GCs in vitro, so that to establish a culture system of buffalo ovarian GCs in vitro. The results showed: the viability of GCs was about 60% after isolation. The growth speed and density of GCs were more dominant as culturing in DMEM medium than that of TCM-199 and DMEM/F12 medium. The GCs began to proliferate after cultured for 24 h, and reached a proliferation peak during cultured for 3 to 5 d. Karyotype analysis displayed that beyond 85% of GCs had normal karyotype in the 1, 3, 5, 7 passage, which did not show that significant difference among these passages. The apoptosis ratio of the 5 and 7 passages of GCs was higher as comparing with the 1 passage (P<0.05, P<0.01). These results indicated that the DMEM medium was more suitable for GCs culturing in vitro, and the GCs could be passaged and keep relatively stable karyotype, but the apoptosis ratio would be elevate with the increase of the passage in vitro culture.

Key words: buffalo; granulosa cell; primary culture; culture medium; proliferation; apoptosis