目的:比较2种人卵巢颗粒细胞分离提纯方法的效果。方法:收集体外受精-胚胎移植(IVF-ET)穿刺取卵后的卵泡液,分别用2种方法分离提取颗粒细胞。A法用红细胞裂解液去除混杂红细胞、酶消化卵泡液中黏液团、Percoll分离液与细胞悬液等体积叠加后离心;B法省略红细胞裂解步骤、舍弃黏液团、增加Percoll分离液用量。比较2种方法获取的颗粒细胞数量、活性、纯度以及实验耗时。结果:2种方法获取的颗粒细胞存活率均>90%,差异无统计学意义(P>0.05)。A法获取的颗粒细胞数量多于B法(P<0.05),但混杂较多细胞碎片,体外培养细胞生长状态欠佳;B法获取的颗粒细胞纯度高,体外培养生长状态良好,且实验耗时较A法短(P<0.05)。结论:2种方法对颗粒细胞存活率无明显影响。B法更为省时高效,有助于建立稳定的颗粒细胞体外培养体系。 Objective: To compare the effects of two human ovarian granulosa cells isolated and purified. Methods: Follicular fluid after IVF-ET in vitro was collected. Two methods were used to separate granulosa cells. A method using red blood cell lysate to remove mixed red blood cells, enzymatic digestion of follicular fluid mucus group, Percoll separation liquid and cell suspension volume superimposed and centrifuged; B omission of red cell lysis step, give up mucus group, increasing the amount of Percoll separation fluid. The number of granulocytes, activity, purity and time taken by the two methods were compared. Results: The survival rates of granulosa cells obtained by the two methods were all> 90%, the difference was not statistically significant (P> 0.05). The number of granulosa cells obtained by method A was more than that of method B (P <0.05), but more cells were mixed with more cell debris. The growth of cells in vitro was poor. The purity of granulosa cells obtained by method B was good, and the growth in vitro was good. Shorter than A method (P <0.05). Conclusion: The two methods have no obvious effect on the survival rate of granulosa cells. B method is more time-saving and efficient, help to establish a stable in vitro culture system of granular cells.