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该研究以鼠尾I型胶原为支架构建了人胚肺成纤维细胞MRC-5三维立体培养模型,探索在此模型中常规培养液和PDGF(platelet-derived growth factor)培养液对MRC-5细胞生长状态的影响,以及这两种培养液对细胞在胶原表面上生长状态的影响。应用此模型初步观察了人巨噬细胞病毒(human cytomegalovirus,HCMV)的感染。结果显示,常规培养液和PDGF培养液均能使细胞相互交错,形成多层的三维立体网状结构。PDGF培养液有利于早期促进细胞呈树突状伸展,常规培养液更有利于细胞在较长时间内维持多层立体的网状结构。常规培养液可促进细胞在胶原表面聚集,PDGF培养液则有利于细胞以单个细胞方式进行迁移。三维胶原体系内生长的MRC-5细胞感染HCMV病毒后,也能呈现明显的细胞病变效应(cytopathic effect,CPE)。由此提示,鼠尾I型胶原与MRC-5细胞构成的三维培养模型,有望进一步应用于细胞生物学、形态学以及病毒感染的研究。
In this study, human embryonic lung fibroblasts MRC-5 three-dimensional culture model was constructed using rat tail collagen I as a scaffold to investigate the effect of conventional culture medium and platelet-derived growth factor (PDGF) The effect of the growth state, and the effect of the two culture solutions on the cell growth on the collagen surface. The model was used to observe the infection of human cytomegalovirus (HCMV). The results showed that the conventional culture medium and PDGF medium can make cells staggered to form a three-dimensional multi-layered network structure. PDGF is conducive to early promotion of cell dendritic elongation, conventional culture fluid is more conducive to cells in a long time to maintain a multi-dimensional three-dimensional network structure. Conventional culture medium can promote the accumulation of cells in the collagen surface, PDGF culture medium is conducive to cells in a single cell migration. MRC-5 cells grown in three-dimensional collagen system also showed obvious cytopathic effect (CPE) after infection with HCMV virus. Thus, the three-dimensional culture model of rat tail collagen type I and MRC-5 cells is expected to be further applied to the study of cell biology, morphology and virus infection.