大鼠卵巢颗粒细胞体外培养方法探讨

Cultured Method Establishment of Granulosa Cells from Rat Ovary

目的建立体外培养大鼠卵巢颗粒细胞分离、体外培养的有效方法。方法收集注射孕马血清促腺激素(PMSG)48h后的大鼠卵泡液,用胰蛋白酶消化法分离纯化颗粒细胞并用含10%胎牛血清的低糖DMEM培养基进行培养。结果得到的卵巢颗粒细胞纯度高,有较好的后续生长增殖能力,可用于后续实验。结论建立大鼠卵巢颗粒细胞体外培养的稳定模型,为颗粒细胞的体外研究提供实验基础。

Objective To obtain and identify cultured granulosa cells from the ovary of rats so as to establish a convenient and stable experimental mothed. Methods Female SD rats were subcutaneously treated with pregnant mare serum gonadotropin （PMSG） . Forty-eight hours after dosing with PMSG, the animals were decapitated and the ovaries were aseptically removed. Granulosa cells were then released by mechanical method, trypsin digestion and low-speed centrifugation separation were used for granulosa cells isohtion. Granulosa cells were diluted andincubated in fresh DMEM-low glucose medium containing 10% of fetal bovine serum. Results Most of cultured cells were ovarian granulosa cells. The growth of cultured cells was good. Conclusion The cultured method of granulosa cells was established stably in this study and was favoring for following experiment.