An integrated microfluidic culture device for quantitative analysis of human embryonic stem cells

被引：80

作者：

Kamei, Ken-ichiro ^{[1
,2
]}

Guo, Shuling ^{[3
,4
]}

Yu, Zeta Tak For ^{[1
,2
,5
]}

Takahashi, Hiroko ^{[1
,2
]}

Gschweng, Eric ^{[3
,4
]}

Suh, Carol ^{[1
,2
]}

Wang, Xiaopu ^{[1
,2
]}

Tang, Jinghua ^{[4
]}

McLaughlin, Jami ^{[3
]}

Witte, Owen N. ^{[1
,3
,4
,7
]}

Lee, Ki-Bum ^{[1
,2
,6
]}

Tseng, Hsian-Rong ^{[1
,2
]}

机构：

[1] Univ Calif Los Angeles, Dept Mol & Med Pharmacol, Los Angeles, CA 90095 USA

[2] Univ Calif Los Angeles, David Geffen Sch Med, Crump Inst Mol Imaging, Los Angeles, CA 90095 USA

[3] Univ Calif Los Angeles, Dept Microbiol Mol Genet & Immunol, Los Angeles, CA 90095 USA

[4] Univ Calif Los Angeles, Eli & Edythe Broad Ctr Regenerat Med & Stem Cell, Los Angeles, CA 90095 USA

[5] Univ Calif Los Angeles, Dept Mech & Aerosp Engn, Los Angeles, CA 90095 USA

[6] Rutgers State Univ, Rutgers Stem Cell Res Ctr, Inst Adv Mat Devices & Nanotechnol, Dept Chem & Chem Biol, Piscataway, NJ 08854 USA

[7] Univ Calif Los Angeles, Howard Hughes Med Inst, David Geffen Sch Med, Los Angeles, CA 90095 USA

来源：

LAB ON A CHIP | 2009年 / 9卷 / 04期

关键词：

DIRECTED DIFFERENTIATION; DEFINED CONDITIONS; LIVING SUBJECTS; REPORTER GENE; GROWTH-FACTOR; MICROENVIRONMENTS; DERIVATION; BACTERIA; GRAVITY; FLOW;

D O I：

10.1039/b809105f

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：555 / 563

页数：9

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