畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (3): 542-548.doi: 10.11843/j.issn.0366-6964.2018.03.010

• 生物技术与繁殖 • 上一篇    下一篇

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

刘迎春1,2, 陈钰萌1,2, 韩哲先1,2, 高峰3*, 周欢敏1,2   

  1. 1. 内蒙古农业大学生命科学学院, 呼和浩特 010018;
    2. 内蒙古自治区生物制造重点实验室, 呼和浩特 010018;
    3. 内蒙古农业大学动物科学学院, 呼和浩特 010018
  • 收稿日期:2017-07-31 出版日期:2018-03-23 发布日期:2018-03-23
  • 通讯作者: 高峰,教授,主要从事动物营养与发育研究,E-mail:gaofeng1994@sina.com
  • 作者简介:刘迎春(1978-),女,内蒙古人,博士,副教授,主要从事干细胞信号通路研究,E-mail:liuyingchun1994@aliyun.com
  • 基金资助:

    国家自然科学基金(31160445)

Induction and Identification of Neural Cell from Mammary Stem Cells in Holstein Cows and the Effect of Prolactin on Its Proliferative Activity

LIU Ying-chun1,2, CHEN Yu-meng1,2, HAN Zhe-xian1,2, GAO Feng3*, ZHOU Huan-min1,2   

  1. 1. College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China;
    2. Key Laboratory of Inner Mongolia Autonomous Region Biological Manufacturing, Hohhot 010018, China;
    3. College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China
  • Received:2017-07-31 Online:2018-03-23 Published:2018-03-23

摘要:

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NSEβ-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。

Abstract:

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 neuron-like 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.

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