大鼠骨髓间质干细胞体外诱导分化为心肌样细胞Cx43通讯连接功能检测  

Study on Cx43 communication function of cardiomyocyte-like cells derived from rat bone marrow mesenchymal stem cells in vitro

作　　者：尹戴佳佳 宋治远[1] 农耀明[1] 

机构地区：[1]第三军医大学西南医院心内科,重庆400038

出　　处：《中国病理生理杂志》2007年第10期1986-1989,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30370585)

摘　　要：目的:探讨大鼠骨髓间充质干细胞(MSCs)体外诱导分化为心肌样细胞Cx43的分布与通讯连接功能状态。方法:取健康SD大鼠骨髓,用5-氮杂胞苷体外诱导培养。取诱导培养2、3、4周的MSCs为Ⅰ、Ⅱ、Ⅲ组,另取急性分离的心肌细胞为对照组,用激光共聚焦技术检测Cx43的分布及平均荧光漂白恢复率。结果:对Cx43在细胞内分布检测发现,随诱导培养时间的延长,细胞内蛋白颗粒密度逐渐增加;诱导培养4周后MSCs内蛋白颗粒密度与对照组比较无显著差异(63.87±12.43,64.87±12.15,P>0.05)。各组细胞平均荧光漂白率的变化趋势与Cx43分布变化相似,即随诱导培养时间的延长,细胞平均荧光漂白率逐渐增加;Ⅰ、Ⅱ、Ⅲ组及对照组分别为19.59%±6.08%、37.17%±3.84%、46.82%±2.69%、49.71%±5.53%,Ⅲ组与对照组比较无显著差异(P>0.05)。结论:大鼠MSCs在体外诱导培养4周后已分化为心肌样细胞,其细胞Cx43的分布与通讯连接功能与正常心肌细胞相似。AIM： To approach the gap junction distribution and communication function of cardiomyocyte - like cells derived from rat bone marrow mesenchymal stem cells （MSCs） in vitro. METHODS： MSCs were isolated by extraction of bone marrow specimens, gradient centrifugation and the adherence of culture plates. MSCs were culture in vitro, treated with 5 - azacytidine and incubated for 24 h. The induced MSCs, which had been incubated for 2, 3 and 4 weeks, were divided into group Ⅰ , Ⅱ and Ⅲ. In addition, the normal cardiomyocyte cells were used as control group. The distribution of connexin 43 （ Cx43 ） and the mean fluorescence redistribution rate were detected in every group with the laser scanning confocal microscope. RESULTS： Cx43 protein grain density in induced MSCs was increased with the lasting of incubation by quantitive analysis of Cx43 distribution. After 4 weeks, the Cx43 protein density in induced MSCs was nonsignificant deviation with control group （63.87 ± 12. 43, 64. 87 ±12. 15, P 〉 0. 05 ）. The diversify tendency of the mean fluorescence redistribution rate was approximation with the result of Cx43 in every group. The results showed that group Ⅰ was 19. 59% ±6. 08%, group Ⅱ was 37. 17% ± 3. 84%, groupⅢ was 46. 82% ±2. 69%, and control group was 49. 71% ± 5.53%. CONCLUSION ： MSCs can be differentiated to cardiomyocyte - like cells, which have been induced and incubated for 4 weeks in vitro. The communicational function of those MSCs is similar to the normal cardiomyocyte cells.

关 键 词：骨髓间质干细胞 大鼠 缝隙接合部 连接蛋白43 心肌样细胞 

分 类 号：R363[医药卫生—病理学]