荷斯坦奶牛乳腺干细胞的成骨诱导及鉴定

Osteogenic induction and identification of mammary stem cells derived from a Holstein dairy cow

背景:课题组已经完成乳腺干细胞向神经细胞和脂肪细胞的诱导,该实验诱导乳腺干细胞分化为成骨细胞,进一步丰富其分化潜能多样性,对乳腺干细胞的研究具有重要意义。目的:探讨荷斯坦奶牛乳腺干细胞定向诱导分化为成骨细胞的潜能。方法:分离纯化的荷斯坦奶牛乳腺干细胞系,由内蒙古生物制造重点实验室提供。取第4代乳腺干细胞加入成骨诱导液培养21 d,采用茜素红染色法和RT-PCR技术进行鉴定。结果与结论:荷斯坦奶牛乳腺干细胞经成骨诱导后,细胞边缘模糊,细胞中有颗粒状钙化结节出现,经茜素红染色呈阳性,诱导后成骨细胞标记性基因Alpl和Bglap呈阳性表达,表明了荷斯坦奶牛乳腺干细胞具有分化为成骨细胞的潜能。

BACKGROUND:The isolated and purified mammary stem cells from Holstein dairy cows have been induced to differentiate into nerve cells,adipocytes and osteoblasts,further enriching their multi-directional differentiation potential,which is of great significance for the study on mammary stem cells.OBJECTIVE:To investigate the osteogenic potential of mammary stem cells from Holstein dairy cows.METHODS:Mammary stem cells from Holstein dairy cows were isolated and purified,which were provided by the Key Laboratory of Biological Manufacturing of Inner Mongolia Autonomous Region.The fourth generation of mammary stem cells were cultured for 21 days in an osteogenic induction medium,and identified by alizarin red staining and RT-PCR.RESULTS AND CONCLUSION:After osteogenic induction,the cell edge was indistinct,and there were some granular calcified nodules in the cells.The induced cells were positive for alizarin red staining.The positive expressions of Alpl and Bglap in the induced osteoblasts were detected,indicating that the mammary stem cells from Holstein dairy cows have the potential to differentiate into osteoblasts.