【摘要】 目的探讨5-氮胞苷体外诱导骨髓间充质干细胞(MSCs)向心肌细胞分化的最佳条件和体外分化的过程。方法体外培养Sprague-Dawley(SD)大鼠的MSCs,传代至P3、P6或P12;用5-氮胞苷诱导,比较不同诱导浓度组、不同诱导时间组和不同体重动物组MSCs的分化率,观察诱导后细胞分化过程中形态学的变化,并且用cardiactroponinI、desmin和α-sarcomericactin抗体鉴定分化的心肌样细胞。结果P12代较P3或P6代骨髓MSCs易向心肌样细胞分化并融合成肌管样结构。经10"mol/L的5-氮胞苷诱导24h后细胞分化率为(27±2.5)%,20"mol/L的5-氮胞苷诱导24h后细胞分化率为(17±2.2)%,其它诱导浓度和诱导时间均不能诱导细胞分化;50g(幼年鼠)体重组细胞分化率为(27±2.8)%,150g(成年鼠)体重组细胞分化率为(15±2.9)%,各组的差异有统计学意义。结论5-氮胞苷诱导MSCs向心肌样细胞分化并融合成肌管样结构,年幼的SD大鼠MSCs易向心肌样细胞分化。更多还原

【Abstract】 [Objective] To explore the most appropriate condition that 5-azacytidine induces bone marrow mesenchymal stem cells(MSCs)to differentiate into cardiomyocytes,and to observe the whole differentiating process of MSCs in vitro.[Methods] MSCs from Sprague-Dawley(SD)rats were cultured and passaged repeatedly to P3,P6,or P12.Experimental groups were divided by various dosage of 5-azacytidine,induce time,and different weights of rats.The continuous morphological changes of the cells were observed after the treatment.Differentiated cardiomyocytes were identified by cardiac troponin I,desmin,and α-sarcomeric actin antibodies immunostaining.[Results] P12 MSCs could more easily be induced and differentiated into cardiomyocyte-like cells than P3 or P6 and subsequently fused to form myotube-like structure.The differentiation ratio was(27±2.5)% by 10 μmol/L 5-azacytidine for 24 hours,but only(17±2.2)% in 20 μmol/L 5-azacytidine for the same time.Differentiation did not happen in any other groups with different inducing concentration and inducing times.The differentiation ratio was(27±2.8)% in 50 g groups compared with(15±2.9)% in 150 g groups.The difference among these groups was statistically significant.[Conclusion] MSCs treated with 5-azacytidine could differentiate into cardiomyocyte-like cells that fused to form myotube-like structure.MSCs from neonatal SD rats could easily differentiate into cardiomyocytes than the old ones.更多还原