DMSO联合高糖体外诱导兔骨髓间充质干细胞分化为胰岛样细胞的研究  

Study of Rabbit Bone Marrow Mesenchymal Stem Cells Differentiating into Islet-Like Cells by DMSO Combined with High Glucose In Vitro

作　　者：于文浩[1] 王亚丹[1] 虎啸[1] 张宁[1] 薛永康[1] 朱邯豫 赵玉玺[1] 王新庄[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002

出　　处：《中国细胞生物学学报》2016年第11期1325-1334,共10页Chinese Journal of Cell Biology

基　　金：国家高新技术研究发展计划(863计划)(批准号:2008AA101010);河南省基础与前沿技术研究计划基金(批准号:092300410081)资助的课题~~

摘　　要：该研究旨在探索二甲基亚砜(dimethyl sulphoxide,DMSO)联合高糖体外诱导日本大耳白兔(Lepus brachyurus)骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)分化为胰岛样细胞的可行性及其调控机制。采用不含血清的DMSO联合高糖诱导P3代兔BMSCs分化为胰岛样细胞。倒置显微镜下观察细胞的形态变化;双硫腙染色和免疫荧光染色检测细胞分化;RTq PCR检测胰岛细胞相关基因[Foxa2(forkhead box A2)、Nestin(neuroepithelial stem cell protein)、Pax6(paired box gene 6)、Pdx-1(pancreatic duodenal homeobox-1)、胰岛素]的表达,并以诱导培养前(0 d)细胞、兔骨髓细胞、P3代BMSCs和胰腺组织中胰岛细胞相关基因表达情况作为参照。结果表明,Foxa2、Nestin和Pax6基因不能作为兔BMSCs向胰岛样细胞诱导分化成功的标志基因。DMSO能够激活Pdx-1基因的表达,促进兔BMSCs分化为可分泌胰岛素的胰岛前体细胞。高糖能够促进兔BMSCs分化为胰岛样细胞,并可显著促进Pdx-1和Foxa2基因的表达。The aim of this study was to explore the feasibility and regulation mechanism of rabbit bone marrow mesenchymal stem cells （BMSCs） differentiating into islet-like cells. Rabbit BMSCs were inducted into islet-like cells by dimethyl sulphoxide （DMSO） combined with high glucose and without serum. The morphological changes of cells were observed under the inverted microscope. The differentiated cells were detected by dithizone （DTZ） staining and immunofluorescence staining. The changes of the expression of islet cell related genes [Foxa2 （forkhead box A2）, Nestin （neuroepithelial stem cell protein）, Pax6 （paired box gene 6）, Pdx-1 （pancreatic duodenal homeobox-1）, Insulin] were detected by RT-qPCR, and the expression of islet cell related genes in cells before induction （0 d）, rabbit bonemarrow cells, P3 BMSCs and rabbit pancreatic tissues were used as references. The results showed that Foxa2, Nestin and Pax6 genes couldnot be used as the marker genes for rabbit BMSCs differentiating into islet-like cells successfully. DMSO could activate the expression of Pdx-1 and promote rabbit BMSCs to differentiate into islet precursor cells which can secrete insulin. High glucose could promote rabbit BMSCs to differentiate into islet-like cells and could significantly promote the expression of Pdx-1 and Foxa2 genes.

关 键 词：兔 骨髓间充质干细胞 二甲基亚砜 高糖 分化 胰岛样细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]