自组装短肽水凝胶支架三维培养环境对骨髓间充质干细胞生物学特性及心肌方向分化的影响

发布时间：2018-12-28 11:33

【摘要】：目的:探究短肽GFS-4自组装形成的水凝胶作为支架材料构建三维微环境对BMSCs生物学特性及向心肌细胞方向诱导分化过程的影响。方法:刚果红染色、红细胞膜裂解实验检测短肽GFS-4自组装效果及对细胞膜是否具有裂解作用;CCK8和AO/EB染色分别检测对BMSCs活性和凋亡的影响;Real-time PCR分析BMSCs诱导分化后MLC-2v、GATA-4基因表达情况。结果:GFS-4自组装后形成致密凝胶,自组装前后对细胞膜无损伤;三维培养环境细胞呈球形生长,细胞活力和凋亡速度均低于二维培养环境。三维培养组在诱导分化过程中的第5天和第7天MLC-2v、GATA-4基因表达均显著高于二维组(P0.05)。结论:短肽GFS-4自组装水凝胶构建的三维微环境延缓了BMSCs的增殖速度和凋亡速度,并促进向心肌方向诱导分化过程中MLC-2v、GATA-4基因的表达。
[Abstract]:Aim: to investigate the effects of hydrogels formed by self-assembly of short peptide GFS-4 as scaffold materials on the biological characteristics of BMSCs and its differentiation to cardiomyocytes. Methods: Congo red staining, erythrocyte membrane splitting assay were used to detect the effect of short peptide GFS-4 on self-assembly and cell membrane cleavage, and the effects of CCK8 and AO/EB staining on the activity and apoptosis of BMSCs, respectively. MLC-2v,GATA-4 gene expression after differentiation induced by BMSCs was analyzed by Real-time PCR. Results: after GFS-4 self-assembly, a dense gel was formed, and the cell membrane was not damaged before and after self-assembly, and the cells in three-dimensional culture environment were spherical, and the cell viability and apoptosis rate were lower than those in two-dimensional culture environment. The expression of MLC-2v,GATA-4 gene in three-dimensional culture group was significantly higher than that in two-dimensional group on the 5th and 7th day of differentiation (P0.05). Conclusion: Three-dimensional microenvironment constructed by self-assembled hydrogel with short peptide GFS-4 can delay the proliferation and apoptosis of BMSCs and promote the expression of MLC-2v,GATA-4 gene in the process of inducing differentiation into myocardium.
【作者单位】：重庆医科大学第一临床学院;重庆医科大学基础医学院分子医学与肿瘤研究中心;义乌市中心医院温州医科大学附属义乌医院;郑州市第七人民医院;
【基金】：国家自然科学基金(31540019,31771101) 重庆市科委自然科学基金(cstc2015jcyj BX0072)资助项目
【分类号】：R542.22

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