摘要
该文通过Tet调控下共表达PDX1与BTC的骨髓间充质干细胞系(PDX1^+BTC^+MSCs),探讨PDX1和BTC共表达对骨髓间充质干细胞分化为胰岛素分泌细胞(IPCs)的效率及成熟度的影响。采用两步法对PDX1^+BTC^+MSCs细胞系诱导分化成IPCs,第一步Dox诱导7天检测到Nestin、CK19表达;第二步再诱导7天后形成DTZ染色阳性的胰岛样结构,Ngn3、Nkx6.1 mRNA水平和PDX1、Insulin、Glucagon的蛋白表达阳性。分化后的IPCs在葡萄糖刺激下能产生胰岛素和C肽,但仍不能达到正常胰岛水平。提示利用Tet-On体系调控PDX1和BTC共表达对骨髓间充质干细胞进行修饰,能有效诱导骨髓间充质干细胞分化为胰岛素分泌细胞,但分化成熟度仍然与天然胰岛细胞功能存在差距。
Replacement of β cells by islet transplantation is a novel therapy for diabetes. Mesenchymal stem cells have been proved to be multipotent. This study evaluate the differentiating ability of rat MSCs into insulin-secreting cells by co-expression of PDXI and BTC. PDX1 is a transcription factor involved in the early endo crine development. Betacellulin （BTC） is a growth factor involved in beta-cell maturation. Co-expression of PDX1 and BTC significantly increased the number of nestin-positive epithelium-like progenitors and islet-like spheroids which differentiated from BMMSCs, and the levels of Insulin and Glut-2 mRNA were elevated significantly. In response to glucose, Pdxl^＋BTC^＋MSCs released insulin and C-peptide, but low compared with normal islets. It is concluded that genetic manipulation of Pdx 1 and BTC by Tet-on system in combination with appropriate differenti- ating culture could induce BMMSCs into the pancreatic lineage in vitro and produce islet-like spheroids that could secrete increased levels of insulin in response to glucose. However, compared with the natural pancreas islet, insulin and C peptide secretion was still insufficient.
出处
《中国细胞生物学学报》
CAS
CSCD
2011年第5期479-484,共6页
Chinese Journal of Cell Biology
基金
国家973前期专项(No.2007CB516811
No.2004CCA01500)
广东省自然科学基金(No.6027540)资助项目~~
关键词
间充质干细胞
PDX1
BTC
分化
胰岛素分泌细胞
mesenchymal stem cells
pdxl
BTC
co-expression
differentiation
insulin-secreting cells
