摘要
本实验通过改良方法使山羊睾丸支持细胞能够在体外快速分离且较长时间培养。将山羊睾丸组织剪碎后经2种混合酶溶液(4 mg/m L胶原酶Ⅳ与0.2%透明质酸酶混合液和0.25%胰酶与10μg/m L DNase Ⅰ混合液)次第消化,利用差速贴壁法纯化细胞悬液中的支持细胞,对培养72 h的细胞经台盼蓝染色鉴定细胞活率,观察支持细胞形态学特征及生长增殖情况,油红O染色、H.E.染色及福尔根染色对支持细胞进行鉴定。结果表明:染色结果观察到在细胞核周围有空泡状结构且有核小体存在,表明本实验分离培养的是山羊睾丸支持细胞,且纯度较高,充分证明了该改良方法可有效的分离山羊睾丸支持细胞并在体外较长时间培养。
A modified method was applied to separate goat sertoli cells fast and culture for the longer term in vitro.The tissue of goat testis was cut into pieces and digested with two types of mixed enzymatic solutions（4 mg/m L collagenase Ⅳ with 0.2% hyaluromidase and 0.25% trypsin with 10 μg/m L DNase Ⅰ） sequentially, sertoli cells were purified from the suspended cells by differential adhesion, and stained with trypan blue to evaluate the viability of the cells which had been cultured 72 h. The morphologic characteristics and proliferation were also observed. Sertoli cells were identified by oil red O, HE and feulgen staining. We observed grains and vacuoles around the nucleu and the nucleosome by the results of staining. These results suggested that the isolated and cultured cells were sertoli cells of goat with high purity and it was fully proved that the modified method could separate goat sertoli cells efficiently and culture for the longer term in vitro.
出处
《中国畜牧杂志》
CAS
北大核心
2014年第21期23-26,68,共5页
Chinese Journal of Animal Science
基金
国家转基因新品种培育重大专项(2011 zx08008-003)
山东省"泰山学者"建设工程专项经费资助
关键词
睾丸
支持细胞
分离
体外培养
山羊
testis
sertoli cell
separate
cultivation in vitro
goat
