联系我们 加入收藏
 
年期检索 高级检索
33
2025年4月5日 星期六
  2016, Vol. 24 Issue (2): 224-232    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
牛肌肉卫星细胞向胰岛素分泌细胞的诱导转分化研究
赵云杰1,魏著英2,李欣欣1,白春玲2,杨慧娣3,李光鹏1
1. 内蒙古大学 哺乳动物生殖生物学与生物技术教育部重点实验室
2. 内蒙古大学
3. 内蒙古医科大学
Trans-differential Induction of Bovine (Bos taurus) Muscle Satellite Cells into Pancreatic Cells
全文: PDF (4957 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
摘要 肌肉卫星细胞是源自中胚层的肌源干细胞,具有增殖分化融合成肌管并形成肌细胞的能力。在体内条件下,肌肉卫星细胞不可能跨胚层分化为内胚层来源的胰腺细胞。本研究从牛(Bos taurus)胎儿肌肉中分离培养得到肌肉卫星细胞,并在体外条件下将其诱导成胰岛素分泌细胞。在诱导过程中,分析了与胰腺发育相关的胰十二指肠同源基因盒1基因(pancreatic and duodenal homeobox 1, PDX1)、神经原素3基因(neurogenin 3, NGN3)、淀粉酶基因(Amylase)和胰岛素基因(Insulin, INS)的动态表达情况。结果表明,PDX1作为决定胰腺分化发育和胰岛功能的主要调节基因,在诱导分化的第2天能够检测到其mRNA的表达,在第3天表达量达到了最大,第4天以后维持在较低水平。NGN3是内分泌细胞形成非常重要的转录因子,是能够将胰岛素分泌到胞外的关键基因,其mRNA在分化诱导的第3天开始表达,第4天达到最高水平,之后逐渐下降。Amylase是胰腺发育中细胞外分泌相关的基因,Amylase mRNA在第6天才开始表达,8 d后达到最高。INS mRNA表达量在11~12 d最高,酶联免疫吸附分析(enzyme-linked immunosorbent assay, ELISA)发现,诱导形成的胰腺细胞能够正常分泌胰岛素,培养液中胰岛素的浓度在诱导的11~12 d达到最高值。向培养液中添加葡萄糖后,可使细胞的胰岛素分泌量显著提高,说明诱导得到的细胞类似于成体胰岛的功能,其胰岛素分泌受外界葡萄糖浓度的调节。上述结果表明,肌肉卫星细胞可以被诱导转分化为胰岛素分泌细胞,并对培养环境中的葡萄糖刺激做出应答反应。本研究结果为进一步的研究和临床应用提供基础资料。
服务
把本文推荐给朋友
加入我的书架
加入引用管理器
E-mail Alert
RSS
作者相关文章
赵云杰
魏著英
李欣欣
白春玲
杨慧娣
李光鹏
关键词 肌肉卫星细胞定向分化胰腺细胞胰岛素分泌    
Abstract:Muscle satellite cells are potential myogenic cells capable of self-renewal and with potential multi-directional differentiation. Under normal circumstances, these cells are in resting state, but they can proliferate, differentiate and integrate into myotubes and form muscle cells. However, in vivo, these mesoderm originated satellite cells can not trans-differentiate into endoderm-derived insulin-producing cells. In this study, a muscle satellite cell line was isolated, cultured, identified and established from a bovine (Bos taurus) fetal muscle tissues. The muscle satellite cells were then induced to differentiate into insulin-producing cells. During pancreatic induction process, the genes associated with pancreatic development such as pancreatic and duodenal homeobox 1 (PDX1), neurogenin 3 (NGN3), Amylase and Insulin (INS) were analyzed. PDX1, an important transcription factor in differentiation of pancreatic islets, started to express after induction 2 d, reached to the highest level after 3 d, and kept lowest level after 4 d. NGN3 was an important transcription factor in the formation of endocrine cells, and was a key gene that controlled insulin secretion to extracellular, which the mRNA started to express after induction 3 d, reached to the highest level after 4 d and then declined. As a pancreatic extracellular secretion associated gene, Amylase mRNA appeared after 6 d and reached to the maximum after 8 d. INS expressed from 1st day and reached to the maximum at 11~12 d. Enzyme-linked immunosorbent assay (ELISA) of the culture media showed that the concentration of insulin secreted by the cultured cells reached to the maximum at 11~12 d. The insulin secretion was significantly increased in the glucose added culture medium, and glucose concentration regulated insulin secretion in the cultured cells, which was similar to adult islet fuction. In conclusion, the bovine muscle satellite cells could be trans-differentiated into pancreatic cells, and make response to glucose stimulation. The study provides basic data for further research and clinical application.
Key wordsMuscle satellite cells    Directed differentiation    Pancreatic cells    Insulin secretion
收稿日期: 2015-07-13      出版日期: 2015-12-29
基金资助:草食家畜干细胞研究
通讯作者: 魏著英     E-mail: weizhuying2008@126.com
引用本文:   
赵云杰 魏著英 李欣欣 白春玲 杨慧娣 李光鹏. 牛肌肉卫星细胞向胰岛素分泌细胞的诱导转分化研究[J]. , 2016, 24(2): 224-232.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2016/V24/I2/224
 
版权所有 © 2014 《农业生物技术学报》编辑部   京ICP备11035905号-3
地址:北京市海淀区圆明园西路2号中国农业大学生命科学楼1053室 邮编:100193
电话:010-62733684 传真:010-62731615 E-mail: nsjxb@cau.edu.cn