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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。
Objective: To investigate whether Tbx18 can successfully transfect adipose-derived stem cells and differentiate adipose-derived stem cells into cardiomyocytes. METHODS: Rabbit adipose derived stem cells derived from the inguinal fat of Japanese eared rabbits were isolated and cultured. Adipose-derived stem cells were transfected with adenoviral vector carrying Tbx18. Differentiation into cardiomyocytes was induced after differentiation. At the same time, Adenovirus group and untransfected group as a control. Transfection efficiency was detected by flow cytometry. Smooth muscle actin α-SMA was detected by immunofluorescence method. The expression of TNTN2 was detected by real-time quantitative PCR. Results: Fluorescent expression was observed under fluorescence microscope after transfection for a long time. Transfection efficiency of flow cytometry was about 70%. After differentiation, the expression of α-SMA and TNNT2 appeared in ADSCs. CONCLUSION: Tbx18 can be successfully transfected into adipose-derived stem cells and can be stably expressed in cells. Tbx18 can induce adipose-derived stem cells to differentiate into cardiomyocyte-like cells.