鸡睾丸细胞体外分离培养初探  

Chicken testicle cells separation and culture

作　　者：李碧春[1] 周冠月[1] 吴洪[1] 孙思宇[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2007年第1期21-25,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(30371031)

摘　　要：取孵化15d鸡胚及出壳7d内和7d后雏鸡的睾丸，分别采用二酶二步消化法（胶原蛋白酶＋胰蛋白酶）和三酶二步消化法（胶原蛋白酶＋透明质酸酶／胰蛋白酶）分离睾丸细胞。结果表明：同一时期睾丸细胞的分离效果，二酶二步消化法与三酶二步消化法差异显著；不同时期采用二酶二步消化法分离睾丸细胞，孵化15d的分离效果显著优于出壳7d内，极显著优于出壳7d后，出壳7d内极显著优于出壳7d后。Percoll离心法提纯结果表明：密度梯度为27％～35％的条带中睾丸细胞纯度较高。在体外无饲养层的条件下分别培养经提纯与未经提纯的睾丸细胞，结果表明未经提纯的细胞具有较高的存活力。The separation methods for both of collagenase typel ＋hyaluronidase/TE and collagenase typel＋TE＋TE with different stage of embryo development were studied. The results showed that： there was significant difference between the method 1 and method 2 at the same stage; The amount of testicle cells purified from different stage showed significant or very significant difference with the method 2. The result of Percoll centrifugal was that in the 27%-35% density cincture, the numbers of testis eells were highest. Culturing the eells without feeder layer in vitro showed that the viability of unpurified testis cells was better than purified testicle cells.

关 键 词：鸡 精原干细胞 分离提纯 培养效果 

分 类 号：S831.3[农业科学—畜牧学]