摘要
目的构建高通量三维细胞共培养芯片,模拟体内微环境,实现多种细胞共培养.方法借助微流控技术构建高通量三维细胞共培养芯片.通过设计芯片、制作光刻阳膜、制作基片、修剪、打孔、清洗处理,封接制成芯片.将基质胶及稀释后的基质胶分别灌入到基质胶灌注单元、细胞培养室内,37℃凝固,利用荧光标记葡聚糖FITC-Dextran灌注到营养液灌注单元,分别于灌注后15 min、180 min测试芯片弥散情况.将膀胱癌T24细胞、纤维细胞、内皮细胞接种到芯片中,培养24h,观察细胞生长情况.结果制作成的高通量三维细胞共培养芯片,大小为5 cm×5 cm×1 cm.芯片包括营养液灌注单元、生物胶灌注单元和细胞培养单元以及微桥连接结构.显微镜下观察FITC-Dextran弥散至整个芯片,3种细胞在该平台生长状态良好.结论本研究实现了4组3种细胞在微小尺度下三维共培养,为进一步研究体内微环境以及以后的药物筛选提供了良好生物平台,将会成为一种新的细胞培养方式.
Objective To fabricate a high throughput three-dimensional cell co-culture chip to simulate microenvironment in vitro.Methods With microfluidic technology,the authors designed and fabricated high throughput three-dimensional cell co-culture chips.The chip was made through photolithography and soft-lithography techniques including spin coating negative photoresist,trimming,punching and cleaning.Then,the matrix and diluted matrix were injected into the matrigel unit and cell culture unit respectively.Once clotting at 37 degrees Celsius,the unit dispersion was observed by using FITC-Dextran after 15min,180min.In this study,human bladder cancer cells(T24),endothelial cells,fibroblasts were inoculated into the chip and cultured for a period of time to observe cell growth.Results The Chip was consisted of two layers,which size was 5cm×5cm×1cm.The chip includes nutrition unit,cell culture chambers,matrigel channel units and microbridge.Under the microscope,the FITC-Dextran was diffused into the whole chip.The cells grew well on the platform.Conclusions The cells co-cultured in three-dimension at the microscale,provided a good biological platform for further study of the in vivo microenvironment and subsequent drug screening.This would become a new cell culture method.
作者
王云超
刘青
牛海涛
门同义
张晓明
杨吉伟
沈彬
王建宁
Wang Yunchao;Liu Qing;Niu Haitao;Men Tongyi;Zhang Xiaoming;Yang Jiwei;Shen Bin;Wang Jianning(Department of Urology,the First Affiliated Hospital of Shandong First Medical University,Jinan,Shandong,250014,China;Department of Ultrasound,Shandong Provincial Hospital Affiliated to Shandong First Medical University,Jinan,Shandong,250021,China;Department of Urology,The Affiliated Hospital of Qingdao University,Qingdao,Shandong,266003,China)
出处
《泌尿外科杂志（电子版）》
2020年第1期17-23,共7页
Journal of Urology for Clinicians(Electronic Version)
关键词
高通量
微流控芯片技术
三维细胞共培养
肿瘤微环境
High throughput
Microfluidic chip
Three-dimensional cell co-culture
Tumor microenvironment
