5-aza-dC体外诱导兔骨髓间充质干细胞分化为胰岛细胞的研究

doi:10.11843/j.issn.0366-6964.2014.09.022

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (9): 1538-1543.doi: 10.11843/j.issn.0366-6964.2014.09.022

5-aza-dC体外诱导兔骨髓间充质干细胞分化为胰岛细胞的研究

吕婧玉^1,2，许晓婷¹，陈晓佩¹，翟明胜¹，冯建昆¹，于文浩¹，王新庄^1*

（1.河南农业大学牧医工程学院，郑州 450002； 2.河南省动物卫生监督所,郑州 450008）

收稿日期:2014-01-29 出版日期:2014-09-23 发布日期:2014-09-23
通讯作者: 王新庄，教授，主要从事动物细胞与胚胎工程研究
作者简介:吕婧玉（1986-），女，河南三门峡人，助理兽医师，硕士，主要从事动物细胞工程研究,E-mail：jingyu0425@aliyun.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

Differentiation of Rabbit Bone Mesenchymal Stem Cells into Islet Cells by 5-Azacytidine in vitro

LYU Jing-yu^1,2，XU Xiao-ting¹，CHEN Xiao-pei¹，ZHAI Ming-sheng¹，FENG Jian-kun¹，YU Wen-hao¹，WANG Xin-zhuang^1*

(1.College of Animal Husbandry and Veterinary，Henan Agricultural University，Zhengzhou 450002,China;2.Henan Animal Health Inspection Institute,Zhengzhou 450008,China)

Received:2014-01-29 Online:2014-09-23 Published:2014-09-23

摘要/Abstract

摘要：

为探讨DNA甲基化水平变化与BMSCs分化的关系，采用MTT法检测不同浓度的DNA甲基化抑制剂5-aza-dC作用不同时间对细胞活力的影响,用1.0 μmol•L^-1的5-aza-dC处理BMSCs，并联合DMSO、尼克酰胺诱导BMSCs向胰岛细胞分化，与未作DNA酶甲基化抑制剂处理的DMSO法诱导组进行双硫腙染色对比、胰岛关键调控基因PDX-1的免疫细胞化学荧光染色鉴定对比。研究表明，0.2~1.0 μmol•L^-1 5-aza-dC为处理兔BMSCs合适的浓度，1.0 μmol•L^-1 的5-aza-dC与DMSO、尼克酰胺诱导剂的联合使用，能够促进胰岛调控关键基因PDX-1的表达，建立了BMSCs向胰岛细胞分化的较为高效的诱导体系。揭示了甲基化程度影响BMSCs向胰岛细胞的分化，低甲基化程度促进胰岛特异基因PDX-1的表达。

Abstract:

To explore the relationship between the changes of DNA methylation level and the differentiation of BMSCs，the effect on cell viability of DNA methylation inhibitors (5-aza-dC) in different concentrations at different times was detected by MTT method.The cells were treated with 5-aza-dC at the concentration of 1.0 μmol•L^-1 and differentiate into islet cells combining with DMSO and nicotinamide，which was compared with the induction group that was not treated with 5-aza-dC with Dithizone staining method and the immunofluorescent staining method in the key regulatory genes of pancreatic islet (PDX-1).The results showed that the proper concentration of 5-aza-dC for the treatment of rabbit BMSCs ranges from 0.2 to 1.0 μmol•L^-1.The combine use of 5-aza-dC at the concentration of 1.0 μmol•L^-1 and DMSO，nicotinamide could promote the expression of the key regulatory genes of pancreatic islet (PDX-1) and established an efficient system for the differentiation of BMSCs into islet cells.It reveals that the degree of methylation could influence the differentiation of BMSCs into islet cells and the low degree of methylation could promote the expression of specific gene of pancreatic islet (PDX-1).

中图分类号:

吕婧玉，许晓婷，陈晓佩，翟明胜，冯建昆，于文浩，王新庄. 5-aza-dC体外诱导兔骨髓间充质干细胞分化为胰岛细胞的研究[J]. 畜牧兽医学报, 2014, 45(9): 1538-1543.

LYU Jing-yu，XU Xiao-ting，CHEN Xiao-pei，ZHAI Ming-sheng，FENG Jian-kun，YU Wen-hao，WANG Xin-zhuang. Differentiation of Rabbit Bone Mesenchymal Stem Cells into Islet Cells by 5-Azacytidine in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(9): 1538-1543.

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5-aza-dC体外诱导兔骨髓间充质干细胞分化为胰岛细胞的研究

Differentiation of Rabbit Bone Mesenchymal Stem Cells into Islet Cells by 5-Azacytidine in vitro

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