稳定表达牛催乳素基因的小鼠乳腺上皮细胞系的建立  

The Establishment of a Mouse Mammary Epithelial Cell Line in Which Bovine Prolactin Can be Stably Transcribed

作　　者：方彧聃[1,2,3] 何佳平[1,2] 张斯敏[1,2] 王娟[1,2,3] 任晓叶[1,2] 张金脉[1,2] 张敬之[1,2,3] 

机构地区：[1]上海市儿童医院上海交通大学附属儿童医院 [2]上海交通大学医学遗传研究所,中国上海200040 [3]卫生部医学胚胎分子生物学重点实验室暨上海市胚胎与生殖工程重点实验室,中国上海200040

出　　处：《生命科学研究》2013年第2期100-105,共6页Life Science Research

基　　金：国家自然科学基金资助项目(81271690);国家重点基础研究发展计划(973计划)项目(2010CB945202);上海市自然科学基金资助项目(11ZR1429900)

摘　　要：在乳腺上皮细胞体外培养时,为了使其状态接近泌乳期,需要在培养基中添加催乳素.由于催乳素价格昂贵,使得乳腺上皮细胞的体外培养成本颇高.建立在体外培养过程中不需要添加催乳素蛋白的乳腺上皮细胞系,能为在细胞水平研究乳腺细胞相关基因提供诸多方便.利用慢病毒载体的整合特性,建立稳定整合了牛催乳素cDNA(bPRL)表达盒的小鼠乳腺上皮细胞系(HC11细胞系).经10代次以上的传代以后,通过定量PCR检测,证明平均每个细胞中含有2.6个外源的bPRL基因,其表达量为HC11细胞中管家基因β-肌动蛋白(β-actin)表达量的14%左右.另外,先前的研究结果表明催乳素能在HC11和泌乳期的小鼠乳腺上皮组织中有效促进山羊β-酪蛋白启动子启动外源基因的表达.之后的实验证实整合了催乳素基因的HC11细胞(bPRL-HC11细胞系)也有此功能.因此,bPRL-HC11细胞系可以为体外研究乳腺生物反应器提供良好的细胞模型.The addition of prolactin is needed for mammary epithelial cell culture in order to maintain its lactogenic feature, though it is costly. Therefore, establishing a prolactin-expressing mammary epithelial cell line might be able to solve this huddle while maintains its lactogenic feature in culture. The establishment of a prolactin-expressing mammary epithelial cell line via Lentiviral vector mediation to integrate a bovine prolactin eDNA expression cassette （bPRL） into the mouse mammary epithelial （HC11） cell line was reported here. Using qPCR analysis, it was demonstrated that the bPRL-expressing cell line （bPRL-HC11） bore with 2.6 copes of bPRL gene/cell, as a whole. The cells transcribed bPRL mRNA efficiently which was accounted for 14% of β-actin mRNA detected by qRT-PCR. In addition, based on our previous finding of prolactin could enhance the transgene expression driven by a goat β-casein promotor both in HCll cells and in murine mammary gland. And the bPRL-expressing HC11 cells could also appear the same function. There-fore, the bPRL-HCll cell line we generated can be a good cell model for the study of mammary gland bioreactor in vitro.

关 键 词：HC11细胞系 催乳素 慢病毒载体 乳腺特异性基因 乳腺生物反应器 

分 类 号：Q291[生物学—细胞生物学]