摘要
以新生牛作为实验动物,取睾丸进行制样组织学观察,并将睾丸消化成单细胞悬液,对生精上皮细胞进行分离纯化、细胞鉴定及冷冻保存,研究不同分离纯化及冷冻保存方法对睾丸细胞的影响。结果表明,新生牛曲细精管中含有精原干细胞和支持细胞,差速贴壁法能有效分离两种细胞;精原干细胞鉴定,AKP、C-kit、OCT-4均呈阳性;支持细胞鉴定,油红O、波形蛋白均呈阳性。精原干细胞冷冻保存以10%的二甲基亚砜为冷冻剂的效果最好,支持细胞冷冻保存以10%乙二醇和0.1 mmol.L-1海藻糖组合效果最好。
Take the new neonatal cattle as the experimental animals,take the bovine testiculars to carry on the sample preparation cytology observation and the bovine testiculars were digested into single cell suspension.The epithlium of sunrex was taken to isolate and purify,and the cells were cryopreserved after identification.Then,the effect of the bovine testicular cells by using different purification and cyopre-servation methods were studied.The results showed that： neonatal bovine seminiferous tubules contained spermatogonial stem cells and sertoli cells,differential adherence selection method can effectively separate these two types of cells.AKP,C-kit,and OCT-4 were positive in identification of spermatogonial stem cells;oil red O and vimentin were positive in identification of sertoli cells.Medium supplemented with 10% DMSO was the optimal choice for spermatogonial stem cells cryopreservation.Medium supplemented with 10% of ethylene glycol and 0.1 mmol.L-1 trehalose was the optimal choice for sertoli cells cryopreservation.
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2010年第2期96-100,共5页
Journal of Northeast Agricultural University
基金
黑龙江省农业科学院博士后基金(LRB04-185)
关键词
精原干细胞
支持细胞
分离纯化
冷冻保存
spermatogonial stem cell
sertoli cell
isolation and purification
cryopreservation
