Tbx18 能使兔脂肪干细胞向心肌样细胞分化

目的:研究Tbx18是否能成功转染脂肪干细胞并使脂肪干细胞向心肌细胞分化。方法:分离培养来源于日本大耳兔腹股沟部脂肪的兔脂肪干细胞,用搭载有Tbx18的腺病毒载体转染脂肪干细胞,诱导分化后检测向心肌细胞的分化情况,同时将转染了含GFP的腺病毒组与未转染组作为对照。采用用流式细胞仪检测转染效率,采用免疫荧光法检测平滑肌肌动蛋白α-SMA,采用实时定量PCR法检测兔肌钙蛋白TNNT2的表达。结果:转染后荧光显微镜下可观察到荧光表达,且持续时间较长。流式细胞仪检测转染效率为70%左右;诱导分化后,脂肪干细胞内出现了α-SMA和TNNT2的表达。结论:Tbx18可成功转染入脂肪干细胞,且能在细胞内稳定表达;Tbx18可诱导脂肪干细胞向心肌样细胞分化。

Tbx18 can Induce Adipose Tissue-derived Mesenchymal StemCell to Differentiation into Cardiomyocyte-like Cells

Objective:To observe if Tbx18 can transfer adipose tissue-derived mesenchymal stemcell successfully and induce it to differentiation into cardiomyocyte-like cells.Methods:Rabbit adipose tissue-derived mesenchymal stem cells were obtained from inguinal adipose of Japanese rabbit. ADSCs were isolated from the subcutaneous adipose tissue of the rabbit, and the third generations were induced to differentiate by transferring Tbx18. After 1 week, the cell differentiation potential was detected. The stem cells transferred with GFP and not transferred anything served as control group. Flow cytometer was used to detect the transfection efficiency. Immunofluorescence was used to detect the positive expression rate of alpha-SMA in ADSCs. Real time RT-PCR was performed to investigate mRNA level of TNNT2.Results:After transfection, the fluorescence could be seen by the fluorescence microscope, and it last long. The flow cytometer shows the transfection efficiency is about 70%; After induced, the ADSCs start to appear alpha-SMA and TNNT2.Conclusion:Tbx18 can be transfected in to ADSCs, and express steadily. Tbx18 can induce adipose tissue-derived mesenchymal stem cell to differentiation into cardiomyocyte-like cells.