微流控芯片细胞动态培养装置的设计与制作
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摘要
基于微流控芯片的细胞培养能有效模拟细胞在体生存微环境,为实现微流控芯片内细胞的长期动态培养及实时观察,设计并制作了一种微流控芯片细胞动态培养装置。首先,采用分体式设计,利用SolidWorks分别构建培养箱、控制箱和载泵箱的三维模型,根据性能要求设计控制系统并配备硬件。接着,利用Comsol软件中的焦耳热模块对加热器ITO玻璃进行热力学仿真,通过分析温度场验证可行性。然后,搭建培养装置,进行调试,并检验其稳定性。最后,进行细胞培养实验。采用分体式设计可有效减小培养箱体积,实现与显微观测系统的兼容;培养装置对温湿度的控制稳定性良好;细胞生长状态良好,生长曲线呈"S"形,细胞存活率达到95%以上。该分体式微流控细胞动态培养装置可长期为细胞培养提供所需环境,并可进行实时观察,满足设计要求。
Cell culture is the basis of cell research.In order to realize long-term dynamic culture and real-time observation of cells in a microfluidic chip,a microfluidic cell dynamic culture device was designed and fabricated.First,based on the "split" arrangement idea,a three-dimension model of the culture box,control box,and pump box were set up using the software,SolidWorks;hardware and software were designed taking the functional demands into consideration.Second,based on the Comsol software platform,the method of Joule heating was used to analyze the thermodynamic characteristics of the device's heater,namely,ITO glass;the temperature distribution was then obtained from the simulation results.Third,the entire device model was established and its stability was tested.Finally,cell culture experiments were performed.The split design can effectively reduce the volume of the incubator and achieve compatibility between the incubator and the microscopic observation system.In addition,the temperature and humidity of the device are stable,cell growth is in the "S" trend,and the cell survival rate is more than 95%.The split-body cell dynamic culture device offers a satisfactory environment condition for cell culture,in which cells can be observed in real time.As such,the device can satisfy relevant design requirements.
Cell culture is the basis of cell research.In order to realize long-term dynamic culture and real-time observation of cells in a microfluidic chip,a microfluidic cell dynamic culture device was designed and fabricated.First,based on the "split" arrangement idea,a three-dimension model of the culture box,control box,and pump box were set up using the software,SolidWorks;hardware and software were designed taking the functional demands into consideration.Second,based on the Comsol software platform,the method of Joule heating was used to analyze the thermodynamic characteristics of the device's heater,namely,ITO glass;the temperature distribution was then obtained from the simulation results.Third,the entire device model was established and its stability was tested.Finally,cell culture experiments were performed.The split design can effectively reduce the volume of the incubator and achieve compatibility between the incubator and the microscopic observation system.In addition,the temperature and humidity of the device are stable,cell growth is in the "S" trend,and the cell survival rate is more than 95%.The split-body cell dynamic culture device offers a satisfactory environment condition for cell culture,in which cells can be observed in real time.As such,the device can satisfy relevant design requirements.
引文
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[18] LI J M,WEI J,LIU Y C,et al..A microfluidic design to provide a stable and uniform in vitro microenvironment for cell culture inspired by the redundancy characteristic of leaf areoles[J].Lab on a Chip,2017,17(22):3921-3933.
[2]宋鸿,韩云竹,蔡国徽.细胞培养技术的研究进展[J].遵义医学院学报,2014,37(5):549-554.SONG H,HAN Y ZH,CAI G H.Research progress of cell culture techniques in scientific Fields[J].Journal of Zunyi Medical University,2014,37(5):549-554.(in Chinese)
[3]王洪成,朱丽,杨利军,等.基于微流体脉冲惯性力驱动虾卵细胞的液滴包裹[J].光学精密工程,2015,23(10):286-290.WANG H CH,ZHU L,YANG L J,et al..Droplet-based encapsulating of shrimp oocyte actuated by microfluidic pulse inertial force[J].Opt.Precision Eng.,2015,23(10):286-290.(in Chinese)
[4]刘冲,刘涛,魏娟,等.细胞三维动态培养微器件的设计与制作[J].光学精密工程,2018,26(7):1672-1679.LIU CH,LIU T,WEI J,et al..Design and fabrication of three-dimensional dynamic cell culture micro-devices[J].Opt.Precision Eng.,2018,26(7):1672-1679.(in Chinese)
[5]刘海侠,刘冲,魏娟,等.新型网状通道微流控细胞培养芯片设计与制作[J].纳米技术与精密工程,2017,15(4):273-279.LIU H X,LIU CH,WEI J,et al..Design and fabrication of a novel microfluidic cell culture chip for mesh channel[J].Nanotechnology and Precision Engineering,2017,15(4):273-279.(in Chinese)
[6] LIU J S,ZHANG Y Y,WANG Z,et al..Design and validation of a microfluidic chip with micropillar arrays for three-dimensional cell culture[J].Chinese Journal of Analytical Chemistry,2017,45(8):1109-1114.
[7] NIETO D,MCGLYNN P,DE LA FUENTE M,et al..Laser microfabrication of a microheater chip for cell culture outside a cell incubator[J].Colloids and Surfaces B:Biointerfaces, 2017, 154:263-269.
[8] GIULITTI S,MAGROFUOCO E,PREVEDELLO L,et al..Optimal periodic perfusion strategy for robust long-term microfluidic cell culture[J].Lab on a Chip,2013,13(22):4430-4441.
[9] AU S H,SHIH S C C,WHEELER A R.Integrated microbioreactor for culture and analysis of Bacteria,algae and yeast[J].Biomedical Microdevices,2011,13(1):41-50.
[10]李晓琼,杨春华,邓玉林,等.一种小型化空间实验室用细胞培养装置[P].北京理工大学,CN10560284618A,2016-5-25.LI X Q,YANG CH H,DENG Y L,et al..A miniaturized cell culture device for space laboratory[P]. Beijing University of Technology,CN10560284618A,2016-5-25.(in Chinese)
[11]施镠佳,谭映军,董景新,等.空间细胞培养芯片系统的弹性膜驱动样品更换[J].光学精密工程,2015,23(5):1340.SHI L J,TAN Y J,DONG J X,et al..Elastic membrane actuated sample replacement for space cell culture microchips[J].Opt.Precision Eng.,2015,23(5):1340-1346.(in Chinese)
[12]兰天.空间细胞培养及实时动态显微成像系统关键技术研究[D].北京:北京理工大学,2015.LAN T.Research on key technologies of space cell culture and real-time dynamic microscopic imaging system[D].Beijing:Beijing University of Technology,2015.(in Chinese)
[13]赵明.基于微流控芯片的便携式恒温灌流细胞培养系统的研究[D].哈尔滨:哈尔滨工业大学,2015.ZHAO M.Research on A portable constant temperature perfusion cell culture system based on microfluidic chips[D].Harbin:Harbin Institute of Technology,2015.(in Chinese)
[14] ZHAO L,CHENG P,LI J X,et al..Analysis of nonadherent apoptotic cells by a quantum dots probe in a microfluidic device for drug screening[J].Technical Notes,2009,81:7075-7080.
[15] SUNG J H,SHULER M L.A micro cell culture analogμCCA)with 3-D hydrogel culture of multiple cell lines to assess metabolism-dependent cytotoxicity of anti-cancer drugs[J].Lab on a Chip,2009(10),9:1385-1394.
[16]张玉龙.塑料品种与性能手册[M〗.北京:化学工业出版社,2007.ZHANG Y L.Manual of plastic varieties and properties[M]. Beijing:Chemical Industry Press,2007.(in Chinese)
[17]占今舜,邢月腾,张彬.细胞培养技术的应用研究进展[J].饲料博览,2013(1):8-11.ZHAN J SH,XING Y T,ZHANG B.Progress on the application of cell culture[J].Feed Review,2013(1):8-11.(in Chinese)
[18] LI J M,WEI J,LIU Y C,et al..A microfluidic design to provide a stable and uniform in vitro microenvironment for cell culture inspired by the redundancy characteristic of leaf areoles[J].Lab on a Chip,2017,17(22):3921-3933.
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