摘要
为了寻求一种快速、高效的体外分离、纯化小鼠睾丸间质细胞的方法，试验采用两种酶（胶原酶Ⅳ和O．25％胰蛋白酶）消化法和6个梯度（70％、65％、60％、55％、50％、45％）的Percoll液对小鼠睾丸间质细胞进行分离、纯化，对分离的细胞进行细胞学观察、台盼蓝染色、3β-羟基类固醇脱氢酶（3β-HSD）染色，采用物理方法、胶原酶Ⅳ＋0．25％胰蛋白酶和柠檬酸钠＋KCI对睾丸间质细胞进行消化传代培养。结果表明：胶原酶Ⅳ和0．25％胰蛋白酶限时消化能够获得较高活率（〉90％）的小鼠睾丸间质细胞；经Percoll细胞分离液分离的细胞纯度高达86％，且在34℃、5％CO2条件下培养的细胞生长良好。
To find a high - efficient way to digest and purify mouse leydig cells, two enzymes and 6 gradients of Percoll were used in present experiments. Mouse leydig cells were stained by Typan blue and 3 β- hydroxy steroid dehydrogenase （3β - HSD） to evaluate the viability of cells. Cells subculture was performed with the treatment of physical method, Trypsin and mixture of sodium citrate and potassium chloride, respectively. The results showed that the viability and purity of leydig ceils were up to more than 90% and 86% digested with Trypsin and IV coilagenase and concentrated with percoll. The bettor cultural condition was 34℃ ,5% CO2.
出处
《黑龙江畜牧兽医》
CAS
北大核心
2010年第7期5-7,共3页
Heilongjiang Animal Science And veterinary Medicine
基金
西北农林科技大学“国家大学生创新性实验计划”项目(G2007016)
