An integrated microfluidic culture device for quantitative analysis of human embryonic stem cells
An integrated microfluidic culture device for quantitative analysis of human embryonic stem cells
复制标题
DOI：
10.1039/b809105f
复制
发表时间：
2009-01-01
期刊：
影响因子：
6.1
通讯作者：
Tseng, Hsian-Rong
中科院分区：
文献类型：
--
作者：
Kamei, Ken-ichiro;Guo, Shuling;Tseng, Hsian-Rong
我们已经成功地设计和制造了一个集成的微流控平台，hESC-mu芯片，它能够在半自动化的方式下对单个hESC菌落进行重复性和定量培养和分析。在该设备中，蛇形微通道允许预筛选游离hESC簇，六个单独寻址的细胞培养室允许平行hESC培养，以及顺序的多参数分析。为了实时定量监测hESC的增殖和多能状态，我们构建了内源性OCT4启动子驱动的EGFP敲入hESC细胞系。对几种多能性标记进行芯片免疫分析，以确认hESC菌落保持其多能性。我们的研究首次在微流体环境中很好地表征了hESC培养和分析，以及在同一设备中并行/多参数/实时/自动检测自我更新和分化的概念验证演示。
We have successfully designed and fabricated an integrated microfluidic platform, the hESC-mu Chip, which is capable of reproducible and quantitative culture and analysis of individual hESC colonies in a semi-automated fashion. In this device, a serpentine microchannel allows pre-screening of dissociated hESC clusters, and six individually addressable cell culture chambers enable parallel hESC culture, as well as multiparameter analyses in sequence. In order to quantitatively monitor hESC proliferation and pluripotency status in real time, knock-in hESC lines with EGFP driven by the endogenous OCT4 promoter were constructed. On-chip immunoassays of several pluripotency markers were carried out to confirm that the hESC colonies maintained their pluripotency. For the first time, our studies demonstrated well characterized hESC culture and analysis in a microfluidic setting, as well as a proof-of-concept demonstration of parallel/multiparameter/real-time/automated examination of self-renewal and differentiation in the same device.