摘要
为进一步简化、优化SD大鼠睾丸支持细胞的体外培养条件及鉴定方法,采用三酶两步消化法和差异贴壁法分离、培养、纯化SD大鼠支持细胞,并用HE染色、油红染色、Feulgen染色及透射电镜扫描其超微结构予以鉴定。结果表明:平均0.10g睾丸组织可获得2.5×106个支持细胞,细胞存活率90%以上,体外培养5h后开始贴壁,96h后细胞生长速率下降,其对数生长期为3～7d,整个体外生长周期约为10～15d。对获得的纯化细胞进行鉴定,发现其形态结构及超微结构与支持细胞的形态特征一致。可见,采用三酶两步消化法和差异贴壁法,可达到分离、培养、纯化SD大鼠支持细胞的目的。
The present study was conducted to optimize the in vitro culture conditions and identification method for sertoli cell of SD rat. The sertoli cells from 21 days old male SD rat were isolated, cultured and purified by two-step digestion using three enzymes and differentiated anchoring method. The uhrastructure of sertoli cell was identified using HE, Oil red and Feulgen dyes through TEM scanning. On an average, 2.5×10^6 sertoli cells can be obtained from about 0.10 g of testicle tissue with the survival rate above 90%. The cell began to anchor after 5 hours in vitro culture, and the growth rate of cells decreased after 96 h. The logarithmic growth duration of cell was found to be 3-7 days and in vitro growth duration was 10-15 days. The morphological characteristics and ultrastructure of purified cells observed through TEM revealed it was in accordance with that of sertoli cell. The presented method was found to accurate for studying the sertoli cells.
出处
《广西农业科学》
CAS
CSCD
2010年第3期259-262,共4页
Guangxi Agricultural Sciences
基金
广西自然科学基金项目(桂科自0991045)
广西大学创新课题计划项目(T32011)
关键词
睾丸
支持细胞
体外培养
鉴定
SD大鼠
testicle
sertoli cell
in vitro culture
identification
SD rat
