HIV-Tat-CLIO标记对大鼠骨髓间充质干细胞生物学行为的影响

Effects of HIV-Tat-CLIO on the biological behaviors of marrow-derlved mesenchymal stem cells in rats

目的探讨HIV-Tat-CLIO标记对体外培养的大鼠骨髓间充质干细胞（rMSCs）生物学行为的影响。方法采用差速贴壁法分离培养rMSCs，将其分为HIV-Tat-CLIO标记组和非标记组。MTT比色测定法检测细胞增殖情况，流式细胞仪分析细胞表面标记物、细胞周期和凋亡程度的变化，免疫组织化学方法研究标记后细胞分化情况，并用Transwell实验检测细胞运动迁移能力的改变。结果HIV-Tat-CLIO标记MSCs后，A570波长的光吸收值无明显变化；细胞表面CD34、CD45和CD90分子的表达差异无统计学意义；处于S期的MSCs数量（S％）、DNA合成前期细胞所占百分比（G1％）、增殖指数PrI值（S＋G2M）％以及细胞凋亡情况均无显著改变（P〉0．05）。结论HIV-Tat-CLIO标记对大鼠MSCs生物学行为无明显影响，标记后的干细胞可以正常增殖、分化及运动迁移。

Objective ： To evaluate the effects of HIV-Tat-CLIO on the biological behaviors of marrow-derived mesenchymal stem ceils （rMSCs） in rats. Methods On the basis of the difference of cellular adherent ability, MSCs from primary cultures of rat marrow were isolated and purified by the preplate technique. In the rMSCs labled with HIV-Tat-CLIO or not, the MTT colorimetric assay and transwell as well as flow cytometry （FCM） were used to detect the effects of labeling. Results In experimental group, the photoabsortion values at A570 had no significant change. FCM revealed that the number of rMSCs in S phase （ S% ）, G1% and the PrI （ S ＋ G2M） % wasnot significantly affected. There was also no change on the manifestation of cells facial markers and their capability for differentiation and migration when the cells were labeled with the HIV-Tat-CLIO. Conclusion The HIV-Tat-CLIO incorporation did not affect the capability for cell viability, proliferation, differentiation and migration of rMSCs.