大鼠腺垂体细胞的体外培养和鉴定

目的:探讨大鼠腺垂体细胞体外培养的技术。方法:选用成年雌性SD大鼠,取腺垂体进行单细胞原代培养,动态观察培养细胞在体外生长的情况,并利用免疫细胞化学染色技术进行鉴定。结果:1细胞形态学观察:腺垂体细胞呈圆形,折光性较强,边缘完整清楚有亮晕。培养2～4d,腺细胞仍为圆形,大部分直径减小,可见少量体积大的细胞,散在分布。培养7d以后,细胞开始变平、聚集成细胞簇。2免疫细胞化学染色:对体外培养1d后的成年大鼠垂体细胞进行LH免疫细胞化学染色鉴定。LH阳性细胞体积大,散在分布,数目较少,约占2%～3%;阳性产物位于胞质内,呈棕黄色。结论:1纯采用机械分离也可以获得理想的腺垂体单细胞悬液;2培养1周内的腺垂体细胞外形变化不大,但绝大部分体积变小,有聚集的趋势;3大而散在的细胞为LH阳性细胞,约占腺垂体细胞的2%～3%。本实验为腺垂体细胞的体外研究提供形态学基础。

Culture and identification of rat anterior pituitary cells in vitro

Objective：To find out the best techniques for culture of rat anterior pituitary cells in vitro. Methods：Anterior pituitary of adult SD female rats were isolated for single cell primary culture, cells growing in vitro were observed, immunocytochemistry method was used to identify expression of luteinizing hormone （LH）. Results ： （1）Morphological examination： cells were round and had strong refraction, cells border were unbroken and had bright halo. After culture for 244 days, cells were still round and majority of cells decreased in diameter, meanwhile a few bigger cells remained in diffused distribution. After culture for 7 days, cells turned to flat and gathered. （2）Immunocytochemistry examination： immunocytochemistry method was used to identify expression of LH of rat anterior pituitary cells after 1 day. LH positive cells were big and scattered growing. LH positive cells were less and accounted for 2%-3% approximately. Positive product was brown in cytoplasm. Conclusion： （1）Idea anterior pituitary single cell suspension could be obtained through simple disassociation. （2） No noticeable shape change of anterior pituitary cells after culture for one week were observed but many ceils decreased in volume and have a tendency to gather. （3）LH positive cells which were bigger and scattered growing accounted for 2 % -3 % approximately. This experiment provided morphological evidence for anterior pituitary cell culture in vitro.