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  

Effects of Self-assembling Peptide Hydrogel Scaffolds for Three-dimensional Culture on Biological Behavior and Capability of Myocardium Differentiation in Bone Marrow Mesenchymal Stem Cells

作　　者：赵正德 陈振银[2] 张慧楠[2] 龚剑萍 许少丹 罗忠礼[2] 

机构地区：[1]重庆医科大学第一临床学院,重庆400042 [2]重庆医科大学基础医学院分子医学与肿瘤研究中心,重庆400042 [3]义乌市中心医院温州医科大学附属义乌医院,义乌322002 [4]郑州市第七人民医院,郑州450016

出　　处：《中国生物工程杂志》2017年第11期45-51,共7页China Biotechnology

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

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(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 BMSCs （bone marrow mesenchymal stem cells） at 3D （3- dimensional） culture microenvironment by peptide hydrogel scaffolds using self-assembling peptide GFS-4 on the biological behavior and the process of myocardium differentiation. Methods： The effect of peptides GFS-4 on selfassembling characteristics and the cell membrane disruptive were examined through Congo red staining and erythroeyte membrane lysis. Then, CCK8 and AO/EB staining were used to assess the difference in cell viability and apoptosis level between 2D （2-dimensional） and 3D microenvironment group. And the expression of MLC-2v and GATA--4 gene in the process of myocardium differentiation by Quantitative Real-Time PCR be analyzed, followed by BMSCs cultured at 2D and 3D environment for 3,5,7days. Results ：The self-assembling peptide GFS-4 form a dense gel after 24 hours. There was no harmful for the cell membrane of the peptide before and after selfassembling. BMSCs at 3D culture environment showed that spherical shape, lower cell viability and lower apoptosis. When compared with the 2D culture environment group, the expression of MLC-2v and GATA-4 gene were respectively higher in the 3D environment group at 5 days and 7 days in the process of myoeardium differentiation （P 〈 0.05 ）. Conclusions： The 3D cuhure environment constructed by peptide hydrogel scaffolds delayed BMSCs＇proliferation and apoptosis rate , and also promoted the expression of MLC-2v and GATA-4 gene during the process of myocardium differentiation.

关 键 词：细胞三维培养 自组装短肽 骨髓间充质干细胞 心肌分化 

分 类 号：Q813[生物学—生物工程]