CAJ | 학술논문

目的观察血清不同浓度对乳鼠骨髓间充质干细胞(MSC)诱导分化的影响,检测胰腺相关因子的表达变化.方法建立骨髓间充质干细胞培养方法,通过RT-PCR检测胰十二指肠同源性盒因子-1(PDX-1)和insulin的mRNA的表达变化,及免疫荧光检测PDX-1和insulin蛋白表达情况.结果高血清培养基和细胞因子能诱导MSC增殖分化,与对照组比较,PDX-1 mRNA在20％、25％ FBS组中增高分别为2.71和2.58倍(P＜0.05)；insulin mRNA在20％和25％ FBS组中增高分别为1.83和1.77倍(P＜0.05)；免疫荧光显示在MSC共表达PDX-1和insulin蛋白增强.结论高浓度血清能促进大鼠MSC表达胰腺相关基因,提示MSC需要不同微环境条件的刺激向胰腺细胞分化,为采用干细胞治疗糖尿病提供资料.
목적 관찰혈청불동농도대유서골수간충질간세포(MSC)유도분화적영향,검측이선상관인자적표체변화.방법 건립골수간충질간세포배양방법,통과RT-PCR검측이십이지장동원성합인자-1(PDX-1)화insulin적mRNA적표체변화,급면역형광검측PDX-1화insulin단백표체정황.결과 고혈청배양기화세포인자능유도MSC증식분화,여대조조비교,PDX-1 mRNA재20％、25％ FBS조중증고분별위2.71화2.58배(P＜0.05)；insulin mRNA재20％화25％ FBS조중증고분별위1.83화1.77배(P＜0.05)；면역형광현시재MSC공표체PDX-1화insulin단백증강.결론 고농도혈청능촉진대서MSC표체이선상관기인,제시MSC수요불동미배경조건적자격향이선세포분화,위채용간세포치료당뇨병제공자료.
Objective To investigate the affects of the high concentration of serum on the differentiation of the marrow mesenchymal stem cells (MSCs) in rat toward the islets cells and examine the expression of PDX-1 and insulin.Methods MSCs were cultured with the different serum and the necessary cytokines.The mRNA expressions of PDX-1 and insulin were detected by RT-PCR and their protein expressions were detected by immunofluorescence.Results MSCs showed the spindle after cultured and became the form of polygon after the induced serum.The level of PDX-1 mRNA increased 2.71 fold in the 20％ FBS group and 2.58 fold in the 25％ FBS group.The level of insulin mRNA increased 1.83 fold in the 20％ FBS group and 1.77 fold in the 25％ FBS group.The mRNA expressions of PDX 1 and insulin were significantly increased in the high-concentration serum group compared to the control(P ＜0.05).The protein expressions of PDX-1 and insulin were enhanced after the stimulation of high serum.Conclusions The high concentration of serum could enhance the mRNA and protein expressions of PDX-1 and insulin in the MSCs and induce the differentiation of MSCs into the islets-like cells for the cell-based therapy for the treatment of diabetes.