小鼠 鸡卵巢颗粒细胞分离 培养方法的比较与优化

The Comparison and Optimization of isolation and culture methods of mouse and chicken ovary Granulosa cells

本试验通过分离与培养小鼠、鸡不同等级卵泡颗粒细胞,比较其培养和形态差异,并对提取方法进行优化。试验选取3周龄雌性昆明系小鼠,200日龄的海蓝褐蛋鸡,分别采用优化的方法分离培养卵巢颗粒细胞,通过观察细胞形态、H.E.染色和间接免疫荧光,对比不同种属颗粒细胞提取方法、培养条件及生长状态。结果表明,与采用机械法分离的小鼠卵巢颗粒细胞相比,采用酶消化法分离的鸡不同等级卵泡颗粒细胞贴壁较快、生长速度快、伪足较多。分析得出小鼠与鸡不同等级卵泡颗粒细胞的分离,培养条件均有差异,且对已有提取方法进行优化,所培养的颗粒细胞状态良好。

The experiment was to isolate and culture granulosa cells from mice and chickens,compare the culture and morphological differences of granulosa cells,and optimize the extraction method.3-week-old female Kunming mice and 200-day-old Hy-Line Brown hens were selected to isolate ovarian granulosa cells.The extraction methods,culture conditions and growth state of granulosa cells of different species were compared through observing morphology,HE and immunfluorescence.The results showed that compared with the mouse ovarian granulosa cells separated by mechanical method,the granulosa cells from chicken different grade follicules extracted by enzymatic digestion were characterized by faster adherence,faster growth,and more pseudo-feet.In summary,the separation method and the culture conditions of granulosa cells from mice and hens were different.The extraction method was optimized based on the existing literature,and the granulosa cells grew well.