人肝星状细胞系LX-2激活过程细胞收缩力的实时测量  

Real-time measurement of cell contractile force during activation of human hepatic stellate cell line LX-2

作　　者：王春苒 张帆[1,2] 单宝娟 刘佳琪 祝连庆[1,2] WANG Chunran;ZHANG Fan;SHAN Baojuan;LIU Jiaqi;ZHU Lianqing(Key Laboratory of the Ministry of Education for Optoelectronic Measurement Technology and Instrument, Beijing Information Science & Technology University, Beijing 100192, P.R.China;Overseas Expertise Introduction Center for Discipline Innovation, Beijing Information Science & Technology University, Beijing 100192,P.R.China)

机构地区：[1]北京信息科技大学光电测试技术及仪器教育部重点实验室,北京100192 [2]北京信息科技大学先进光电子器件与系统创新引智基地,北京100192

出　　处：《生物医学工程学杂志》2019年第5期841-849,共9页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(61605011,61875237);科研重点研究培育项目(5211910920);北京市教育委员会科技计划项目(KM201711232004)

摘　　要：肝星状细胞的细胞收缩力在肝脏损伤、炎症和纤维化的过程中起着非常重要的作用。本文提出了一种基于聚二甲基硅氧烷(PDMS)薄微柱阵列测量人肝星状细胞系LX-2细胞收缩力的方法,以实现亚细胞级的细胞收缩力大小、方向、分布及其动态变化的测量。首先,运用二次铸模工艺制备了以玻底培养皿为基底的满足100倍物镜成像工作距离的薄微柱阵列,并对微柱表面进行亲水、压印蛋白处理,再在其上接种细胞。然后,将细胞培养在无胎牛血清(FBS)培养基中以使细胞处于静息态,再通过加入FBS的方法使细胞转变为激活态。通过图像处理技术得到不同时刻微柱顶端发生的位移,结合微柱力学仿真结果,计算得到作用在微柱上的细胞收缩力,并对细胞状态激活前后微柱的受力情况进行了分析。本研究的实验结果表明,静息态时,细胞收缩力最大为20 nN;加入FBS激活细胞后,细胞收缩力最大增强至110 nN。本文所述方法可测量单个LX-2细胞在静息态和激活态时的收缩力,对肝纤维化引发的慢性肝病的病理研究、药物筛选具有重要的意义。The contractile force of hepatic stellate cells plays a very important role in liver damage, hepatitis and fibrosis. In this paper, a method based on polydimethylsiloxane(PDMS) thin micropillar arrays is proposed to measure the contractile force of human hepatic stellate cell line LX-2, which enables dynamic measurement of the subcellular distribution of force magnitude and direction. First, thin micropillar arrays on glass bottom dish were fabricated using two-step casting process in order to meet the working distance requirement of 100× objective lens. After hydrophilic treatment and protein imprint, cells were seeded on the micropillar arrays. LX-2 cells, which were quiesced by growth in serum-free medium, were activated by adding fetal bovine serum(FBS). The deflections of the micropillars were achieved by image processing technique, and then the contractile force of cells exerted on the micropillars was calculated according to mechanical simulation results, and was analyzed under both quiescent and activated conditions. The experimental results show that the average traction force of quiescent cells is about 20 nN, while the contractile force of activated cells increased to 110 nN upon adding FBS. This method can quantify the contractile force of LX-2 cell on subcellular scale in both quiescent and activated states, which may benefit pathology study and drug screen for chronic liver diseases resulted from liver fibrosis.

关 键 词：肝星状细胞系 收缩力 微柱阵列 激活 

分 类 号：R575[医药卫生—消化系统]