基于微流控芯片的无CO2环境下神经细胞与免疫细胞培养技术研究

空间站是空间生命科学研究的重要平台，而空间站难以提供细胞培养所需要的CO₂环境，并且需要可控且自动化的细胞培养方法，因此建立基于微流控芯片的无CO₂细胞培养系统至关重要.本研究从改变培养基成分的初始pH值入手（向培养基中添加外源CO₂或缓冲物质），探寻无CO₂环境下培养细胞的可行性.用光学显微镜镜检细胞生存状态，用MTS法测定细胞活力.结果表明向培养基中添加不同比例的HEPES，在一定浓度的HEPES缓冲液条件下，可实现无CO₂环境下培养人神经相关细胞SH-SY5Y和U87MG以及人免疫相关细胞THP-1，并且向培养基添加3% HEPES可以使细胞至少存活5天，在此基础上又优化了结合微流控芯片培养细胞的条件.

Cell Culture of Nerve Cell and Immune Cell in CO2-Free Environment Based on Microfludic Chip

Space station is important to space biology and it is difficult to provide CO₂ environment,a crucial element to cell culture.Meanwhile,a controllable and automatic method of cell culture is needed to perform the research in space station.Consequently,the establishment of cell culture based on microfluidic chip in a CO₂-free environment is extremely important.Firstly,changed the component of medium (Initial pH of the medium was changed,then adding up exogenous CO₂ into the medium,finally adding up buffer in culture medium),and performed the cell culture in CO₂-free environment (the co-culture of U87 MG and SHSY-5Y and THP-1),then used MTS method to determine the cell viability.Secondly,after adding different proportions of HEPES, performed the cell culture in CO₂-free environment,the cell viability was determinated by MTS method.Finally,according to the optimal proportion of HEPES,the cells were cultured in microfluidic chip.It was concluded that in CO₂-free environment,cells in microfluidic chip would survive in the medium with 3% HEPES for 5 days.