【摘要】 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。更多还原

【Abstract】 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:①Morphological examination:cells were round and had strong refraction,cells border were unbroken and had bright halo. After culture for 2～4 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.②Immunocytochemistry 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:①Idea 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 cells decreased in volume and have a tendency to gather. ③LH 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.更多还原