乳鼠心肌细胞对骨髓基质干细胞分化的影响

The effect of myocytes of neonatal rats on the differentiation of mesenchymal stem cells

目的了解乳鼠心肌细胞对骨髓基质干细胞(MSCs)分化的影响。方法将已传一代的骨髓基质干细胞用Hoechst33258预标记。乳鼠心肌细胞经培养2d后再与骨髓基质干细胞共同培养5d,细胞爬片使用激光扫描共聚焦显微镜技术,经免疫荧光细胞化学鉴定肌钙蛋白-T的表达。流式细胞仪进行骨髓基质干细胞分选及乳鼠心肌细胞周期测定;提取各类细胞总RNA,行逆转录聚合酶链反应技术(RT-PCR)鉴定肌钙蛋白-T(1Tn-T1)的RNA表达。结果共同培养的MSCS表达肌钙蛋白-T及其RNA,而单独培养的MSCS无Tn-T1及其RNA表达;培养1d后乳鼠心肌细胞周期为G0/G176%,S12%,G2/M12%。结论在共同培养下,骨髓基质干细胞向心肌样细胞分化,这种分化不需要细胞与细胞的直接接触。

Objective To evaluate the effect of neonatal rats myocytes on the differentiation of mesenchymal stem cells （MSCs） under cocuhure. Methods MSCs were isolated by their adherence to plastic in vitro and prelabelled by Hoechst 33258 after two generations culture. Cardiomyocytes were derived from one-day-old rats and cultured for 2 days, then they were eocultured with prelabelled MSCs for 5 days. Fluorescence-detivated cell sorting （FACS） was used to extract the prelabelled MSCs and detect the cell cycle of cardiomyocytes. Total RNA was isolated and reverse transcriptase-polymerase chain reaction （RT-PCR） was performed to detect the expression troponin-T1 genes. Immunostaining against troponin-T was performed and detected by Confocal laser scanning microscope （CLSM）. Results FACS identified 76% of the cardiac mymocytcs in the G0/G1 phase, the remaining cells were in S phase（ 12% ）and C2/M phase（ 12% ）. Troponin-T was identified by RT-PCR and immunostaining at the differntiated MSCs. Conclusion MSCs were able to different into cardiomyogenic lineage without direct cell-to-cell contact with cardimyocytes.