荷斯坦奶牛乳腺干细胞的成神经诱导鉴定及催乳素对其增殖活性的影响

摘要

This study investigated the multi-directional differentiation potential of mammary stem cells in Holstein cows and the effect of prolactin on its proliferative activity.The Nissl staining was performed to identify the nerve cells induced from the mammary stem cells in Holstein cows,and the marker genes of neuronal cells were analyzed by RT-PCR.The effect of prolactin on the proliferation of mammary stem cells was detected by MTT.The results indicated that the neuronlike cells and microtubule-like structure were found during induction of mammary stem cells,which was positive by Nissl staining.The marker gene (β-Tubulin Ⅲ and NSE) of nerve cells were positive expression after induction.Prolactin could improve the proliferation of mammary stem cells in Holstein cows,and the optimal proliferation concentration of prolactin to mammary stem cells was at 100 ng · mL-1.%本研究旨在探究荷斯坦奶牛乳腺干细胞的多向分化潜能及催乳素对其增殖活性的影响.采用尼氏染色法对乳腺干细胞向神经细胞分化的情况进行鉴定,利用RT-PCR技术对诱导后的乳腺干细胞进行基因水平检测;MTT法检测催乳素对乳腺干细胞增殖活性的影响.结果表明:荷斯坦奶牛乳腺干细胞经成神经诱导后,细胞中有类似神经元的细胞出现,并且细胞周围出现许多微管样的结构,经尼氏染色呈阳性.诱导后神经细胞标记基因NSE和β-Tubulin Ⅲ呈阳性表达.催乳素刺激可促进乳腺干细胞的增殖,低浓度时乳腺干细胞的活力较好,且100 ng·mL-1为增殖最适浓度.