畜牧兽医学报

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兔骨髓间充质干细胞向胰岛细胞分化的研究

何俊丹1,2,张志帅3,王新庄1*,吕婧玉1,许晓婷1,蒋金航1,翟明胜1   

  1. 1.河南农业大学牧医工程学院,郑州 450002; 2.开封市动物疫病预防控制中心,开封 475004;3.河南省动物卫生监督所,郑州 450008
  • 收稿日期:2012-04-18 出版日期:2013-04-23 发布日期:2013-04-23
  • 通讯作者: 王新庄,教授,E-mail:wangxinzhuang@yahoo.com.cn
  • 作者简介:俊丹(1985-),女,河南许昌人,硕士,主要从事动物细胞工程与临床兽医学研究,E-mail:jundanhe@163.com
  • 基金资助:

    河南省基础与前沿技术研究计划(092300410081);河南农业大学人才引进基金

Study of Inducing Rabbit BMSCs into Islet Cells

HE Jun-dan1,2, ZHANG Zhi-shuai3, WANG Xin-zhuang1*, LV Jing-yu1, XU Xiao-ting1,JIANG Jin-hang1, ZHAI Ming-sheng1   

  1. 1. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China; 2. Kaifeng Animal Disease Prevention and Control Center, Kaifeng 475004, China; 3. Henan Animal Health Inspection Institute, Zhengzhou 450008, China
  • Received:2012-04-18 Online:2013-04-23 Published:2013-04-23

摘要:

旨在探讨体外诱导兔骨髓间充质干细胞向胰岛细胞分化的可行性,本研究采用二甲基亚砜联合高浓度葡萄糖对分离纯化的第3代兔BMSCs进行体外诱导,观察诱导前后细胞形态学变化,用双硫腙染色法鉴定胰岛细胞,用免疫细胞化学法检测胰岛素的表达,用ELISA法检测分泌到培养基中的胰岛素含量。结果,试验组在诱导7 d出现细胞团样集落,10 d获得排列紧密的细胞团,而对照组始终未见细胞团;经双硫腙染色试验组、免疫细胞化学染色试验组均显示阳性,对照组为阴性,ELISA检测试验组7和10 d的胰岛素含量分别为(27.82±0.43)和(27.89±0.25)mU·L-1,与对照组的(23.39±0.20)和(25.71±0.27)mU·L-1比较,差异极显著(P<0.01)。结果表明,二甲基亚砜联合高糖能够在较短的周期内诱导出胰岛细胞。

Abstract:

To explore the method for inducing rabbit bone marrow mesenchymal stem cells(BMSCs) into islet cells in vitro, BMSCs at 3rd generation were induced with dimethyl sulfoxide (DMSO) and high concentration of glucose.The morphological change of cells were observed after inducing. The islet cells were identified by Dithizone staining, the expression of insulin was detected by immunocytochemical staining. The insulin content was detected by ELISA. After induced 7 days,the cells appearred groups of cell mass. After 10 days, groups of cell mass got closely, whereas the control cells did not appeare group of cell mass. After treated with Dithizone(DTZ) and Immunocellulerchemistry (ICC), the induced cells both were positive and the control group both were negative.In ELISA tests,the insulin content of induced group were (27.82±0.43) and (27.89±0.25) mU·L-1 at 7 and 10 days, that of the control group were (23.39±0.20) and (25.71±0.27)mU·L-1. The difference was significant(P<0.01) between induced and control groups. The result showed that DMSO with high concentration of glucose can induce rabbit BMSCs into islet cells in a relatively short period.

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