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。
[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 ceils 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 ceils than P3 or P6 and subsequently fused to form myotube-like structure. The differentiation ratio was （27±2.5）% by 10μmol/L 5- azacytidine for 24 hours, but only （17±2.2）% in 20μmol/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 ceils that fused to form myotube-like structure. MSCs from neonatal SD rats could easily differentiate into cardiomyocytes than the old ones.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2006年第1期33-37,共5页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金资助项目(39970237)
关键词
间充质干细胞
5-氮胞苷
细胞分化
mesenchymal stem cell
5-azacytidine
cell differentiation
