温度对体外培养精原细胞增殖和凋亡的影响  

Influence of temperature on proliferation and apoptosis of spermatogonia cultured in vitro

作　　者：张晓丽[1,2] 高英茂[2] 赵舒武[3] 邴鲁军[2] 

机构地区：[1]山东大学省立医院生殖中心,济南250021 [2]山东大学医学院组织学与胚胎学教研室,济南250012 [3]天津中医药大学组织学与胚胎学教研室,天津300193

出　　处：《解剖学杂志》2008年第6期739-742,共4页Chinese Journal of Anatomy

基　　金：教育部高等学校博士学科点专项科研基金(20010422005)

摘　　要：目的:探讨精原细胞最适培养条件并观察环境温度对体外培养的精原细胞增殖和凋亡的影响。方法:将分离、富集的小鼠精原细胞分别置于不同温度条件下进行体外培养,观察其贴壁、增殖、形态学变化和存活时间;MTT法检测精原细胞活性,Annexin V-FITC/PI染色法检测精原细胞的凋亡。结果:32℃、34℃、37℃条件下均可以看到精原细胞的贴壁、分裂和增殖,39℃培养条件下,精原细胞极少贴壁,基本看不到细胞增殖,细胞很快发生凋亡。随着培养时间延长,精原细胞增殖形成的精原细胞克隆数和精原细胞存活时间明显不同,其中34℃条件下精原细胞增殖最多,存活时间最长。结论:温度对体外培养的精原细胞的增殖和凋亡有明显的影响,34℃为精原细胞体外培养的最适温度。Objective： To find the optimal culture condition and investigate the influence of temperature on proliferation and apoptosis of spermatogonia cultured in vitro. Methods： The isolated and enriched spermatogonia were cultured in incubator at 32℃, 34℃, 37℃ and 39℃. The adherence, morphologic changes and survival time of spermatogonia were observed and recorded under an inverted phase contrast microscope every day. On the 3rd day culture in vitro, survival of spennatogonia was determined by MTT assay, and apoptosis of the spermatogonia was detected by Annexin V FITC/PI staining and ob served under laser confocal scanning microscope. The spermatogonia clone size, spermatogonia survival time, proliferation rate and apoptosis rate were compared in differenl groups. Results： Adherence and proliferation was observed in 32℃, 34℃ and 37℃ groups. In 39℃ group, adherence and proliferation was hardly observed and apoptosis occured quickly. With time prolonging, the numer of spermatogonia clone and survival time obviously changed in different groups. The number of spermatogonia clones cul turcd in 34℃ incubator was the most and the survival time was the longest. Conclusion： Temperature has great influence on spermatogonia and 34℃ is the optimal temperature for spermatogonia culture in vitro.

关 键 词：精原细胞 温度 细胞增殖 细胞凋亡 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]