三维胶原培养模型中mrc-5细胞的生长状态及病毒感染.pdf

– DOI: 10.11844/cj cb.2017.03.0340 中国细胞生物学学报 Chinese Journal of Cell Biology 2017, 39(3): 333 340 技术与方法 三维胶原培养模型中MRC-5细胞的 生长状态及病毒感染 * * 何 兵 陈国敏 曾 毅 (传染病预防控制国家重点实验室, 中国疾病预防中心病毒病预防控制所, 北京102206) 摘要 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。 关键词 人胚肺成纤维细胞; 三维细胞培养模型; 胶原; 巨噬细胞病毒 Growth and Viral Infection of MRC-5 Cells in A Three-Dimensional Collagen Culture Model He Bing, Chen Guomin*, Zeng Yi* (State Key Laboratory of Infection Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China) Abstract In this study, human embryonic lung fibroblasts (MRC-5) were cultured in the three-dimensional (3D) culture model with rat tail tendon collagen type I as a scaffold. The effects of conventional culture media and PDGF (platelet-derived growth factor) media on the growth status of MRC-5 cells were investigated in a 3D culture model. And the effects of the cellular growth status on the surface of 3D collagen matrices with two different culture media were analyzed. The infection of HCMV (human cytomeg