摘要
以新生犊牛睾丸为实验对象,应用组合酶法进行支持细胞分离培养,并研究了冷冻保存后支持细胞的生长特性。结果表明：在细胞分离时,消化睾丸组织,分离曲细精管法所获得的细胞悬液中的有效细胞数高于组织剪碎法。支持细胞体外培养,4 h后开始贴壁,3~4 d铺满培养皿底壁,传代后细胞生长较快,2 d即可增殖一代。HE染色,胞质染色较淡,而细胞核染色较深,呈圆形或椭圆形位于细胞质中央或偏位,核仁明显。采用10%FBS＋10%DMSO的DMEM液做冷冻液,对细胞进行冷冻保存时,支持细胞的复苏率在65%以上。解冻后的支持细胞体外培养,4h开始有细胞贴壁,24h后大部分细胞贴壁,3~4d铺满培养皿底壁。
Newborn calves as experimental subjects, preparation and culture were done ＂after cryopreservation ofnewborn bovine sertoli cells,and characteristics of sertoli cells were observed.The results showed that newborn calf testis seminiferous tubules were solid tissue, spermatogonial stem cells were arranged in the seminiferous tubules of the central or sub- central. Sertoli cells packed , and attached to the base membrane. The number of effective cells through digestion of testicular and isolated seminiferous tubules were more than that in the scissors method. Sertoli cells in vitro, 4 h after adherent , 3-4 d covered dish bottom wall, and passage cells growed rapidly,proliferation of generation twice a day. HE staining, cytoplasmic staining lighter and darker nuclei,were round or oval central or deviation in the cytoplasm,prominent nucleoli. After cryopreservation, the recovery rate was 65 % or more. Sertoli cells after thawing in vitro, adherent cells began at 4 h, the number of adherent cells increased significantly at 6 h,the majority of the cells were adherent 12 h later and covered dish bottom wall in 3-4 days.
出处
《中国草食动物科学》
CAS
2012年第4期21-26,共6页
China Herbivore Science
关键词
犊牛
睾丸支持细胞
体外培养
冷冻保存
newborn bovine
sertoli cells
in vitro culture
cryopreservation
