不同培养模式下奶牛乳腺上皮细胞形态及酪蛋白表达的差异  

Differences of Morphology and Casein Expression of Bovine Mammary Epithelial Cells under Different Culture Models

作　　者：詹康[1] 左晓昕 贡笑笑 陈银银[1] 占今舜[1] 赵国琦[1] 

机构地区：[1]扬州大学动物科学与技术学院,扬州225009

出　　处：《动物营养学报》2015年第7期2241-2247,共7页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：江苏省高校优势学科建设工程资助项目;江苏省科技支撑项目(BE2013387)

摘　　要：本试验旨在研究二维和三维培养模式下奶牛乳腺上皮细胞形态及酪蛋白表达的差异。利用组织块培养细胞来分离奶牛乳腺上皮细胞,利用有限稀释法克隆奶牛乳腺上皮细胞。通过显微镜观察细胞在基质胶中的形态,并用Western blot法检测酪蛋白的诱导表达。结果表明:1)采用组织块培养细胞能够成功分离奶牛乳腺上皮细胞,克隆得到的细胞能稳定传至20代。2)细胞接种12h之后,二维培养的奶牛乳腺上皮细胞并没有形成明显的细胞簇,细胞间也没有相互连接;三维培养的奶牛乳腺上皮细胞形成明显的细胞簇,在基质胶内的层黏连蛋白作用下形成交织网状结构。培养2d细胞间交织网状结构逐渐消失,细胞开始形成单个的成细胞簇,使细胞间更加紧密的相连。3)二维培养奶牛乳腺上皮细胞不能诱导表达酪蛋白,但三维培养细胞可诱导表达κ-酪蛋白。综上所述,三维培养奶牛乳腺上皮细胞能够改变细胞的形态和功能,诱导κ-酪蛋白的表达,三维培养的奶牛乳腺上皮细胞可作为奶牛的泌乳机制研究的体外模型。This research aimed to study the differences of morphology and casein expression of bovine mammary epithelial cells under two- and three-dimensional culture models. Bovine mammary epithelial cells were isolated by tissue mass culture, and were cloned by limiting dilution mehod. Morphology was observed in matrigel using microscopy. The induced casein expression was determined by Western blot method. The results showed as follows： 1 ） Bovine mammary epithelial cells could be successfully isolated by tissue mass culture, and the clones were passaged 20 generations stably. 2 ） After 12 h cultivation, the two-dimensional cultured bovine mammary epithelial cells neither formed apparent clusters nor produced interconnection, however, the three-dimensional cultured cells formed apparent clusters and produced interconnection under the action of laminin in matrigel. The bovine mammary epithelial cells in matrigel started to form network structure and produce interconnection. After 2 d of cultivation, the network structure among cells began to disappear, and single clusters started to appear, which made the connection among cells tighter. 3） The bovine mammary epithelial cells in two-dimensional culture did not express casein, but the expression of K-casein could be induced in three-dimensional culture. It is concluded that the bovine mammary epithelial cells are able to change the morphology and function by threeimensional culture, and can induce the K-casein expression, can be used as an in vitro model for cows＇ lactating mechanism research.

关 键 词：奶牛乳腺上皮细胞 三维培养 酪蛋白 体外模型 

分 类 号：S823[农业科学—畜牧学]