摘要
目的探讨胚胎干细胞是否适合在聚(3-羟基丁酸酯-co-4-羟基丁酸酯)[P(3HB-co-4HB)]上分化生成心肌补片,添加维生素C能否提高诱导率。方法从3只平均体重为37.5 g的清洁级雌性昆明小白鼠胎鼠中提取胚胎纤维状(MEF)细胞,复苏并传代培养小鼠胚胎干细胞(ESC)。小鼠胚胎干细胞来自于上海生命科学院。用悬滴法形成拟胚体后与细胞补片共培养,实验组使用含维生素C的培养基进行培养,对照组使用不含维生素C的培养基进行培养。使用免疫荧光染色和扫描电镜观察心肌细胞,计算诱导分化效率,绘制分化效率随诱导时间的分布图。结果分布图结果显示分化效率随时间逐渐增高,不加维生素C组最高诱导率为17.8%,加维生素C组最高诱导率为71.1%,差异有统计学意义(P<0.05)。结论在P(3HB-co-4HB)生物补片上,ESC可生长、增殖、分化为心肌细胞,添加维生素C可提高诱导率。
Objective To assess the suitability of P（3HB-co-4HB） combined with embryonic stem cells（ESCs） for myocardial patch formation and whether adding vitamin C would improve inductivity or not.Method We extracted mouse embryonic fibrous cell from three clean female white Kunming mouses at a mean body weight of 37.5 grams.We recovered and cultured mouse ESCs.Those mouse embryonic stem cells were obtained from Shanghai Institutes of Biological Sciences.We took pendant-drop method to form embryonic bodies（EBs） and co-cultured them with myocardial patch.The experimental group were cultured in the substate with vitamin C while the control group were cultured in the substate without vitamin C.We immunostained the myocardial patch and observed them by scanning electron microscope.We calculated the differentiation efficiency and mapped the distribution curve of induction time.Results The scattergram showed that the differentiation efficiency increased gradually.The differentiation efficiency of the group with vitamin C was 71.1% and the group without vitamin C was 17.8%.There was a statistical difference between the two groups（P〈0.05）.Conclusion On the biological patch of P（3HB-co-4HB）,ESCs could grow,proliferate,and differentiate into myocardial cell and adding vitamin C into it could improve the differentiation efficiency.
出处
《中国胸心血管外科临床杂志》
CAS
CSCD
2016年第5期496-500,共5页
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery
基金
北京市朝阳区科委社会发展基金(SF1417)
首都医科大学学生科研创新项目(XSKY2015192)~~
