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.
出处
《生命科学研究》
CAS
CSCD
北大核心
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)
