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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(P<0.05)。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。
OBJECTIVE: To investigate the effect of three-dimensional microenvironment on the biological characteristics of BMSCs and the direction of cardiomyocyte differentiation induced by self-assembled hydrogel GFS-4 as a scaffold material. METHODS: Congo red staining and erythrocyte membrane lysis assay was used to detect the effect of GFS-4 self-assembly and cell membrane lysis. CCK8 and AO / EB staining were used to detect the effect on BMSCs activity and apoptosis. Real-time PCR analysis of BMSCs Induction of differentiation after MLC-2v, GATA-4 gene expression. Results: GFS-4 formed compact gel after self-assembly, and had no damage to the cell membrane before and after self-assembly. The three-dimensional culture environment showed spherical growth, cell viability and apoptosis rate were lower than the two-dimensional culture environment. The gene expression of MLC-2v and GATA-4 in three-dimensional culture group was significantly higher than that in two-dimensional group (P <0.05) on the 5th and 7th day during differentiation. CONCLUSION: The three-dimensional microenvironment of GFS-4 self-assembled hydrogel can delay the proliferation and apoptosis of BMSCs, and promote the expression of MLC-2v and GATA-4 in the direction of myocardial differentiation.