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

Ken-ichiro Kamei; Shuling Guo; Zeta Tak For Yu; Hiroko Takahashi; Eric Gschweng; Carol Suh; Xiaopu Wang; Jinghua Tang; Jami McLaughlin; Owen N. Witte; Ki-Bum Lee; Hsian-Rong Tseng

doi:10.1039/B809105F

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

Ken-ichiro Kamei,‡^ab Shuling Guo,‡^cd Zeta Tak For Yu,‡^abe Hiroko Takahashi,^ab Eric Gschweng,^cd Carol Suh,^ab Xiaopu Wang,^ab Jinghua Tang,^d Jami McLaughlin,^c Owen N. Witte,*^acdg Ki-Bum Lee*^abf and Hsian-Rong Tseng*^ab

Author affiliations

* Corresponding authors

^a Department of Molecular & Medical Pharmacology, University of California, Los Angeles, USA
E-mail: hrtseng@mednet.ucla.edu

^b Crump Institute for Molecular Imaging, David Geffen School of Medicine, University of California, Los Angeles, USA

^c Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, USA

^d The Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, USA
E-mail: owenw@microbio.ucla.edu

^e Department of Mechanical and Aerospace Engineering, University of California, Los Angeles, USA

^f Department of Chemistry & Chemical Biology, Institute for Advanced Materials, Devices and Nanotechnology, The Rutgers Stem Cell Research Center, Rutgers, The State University of New Jersey, Piscataway, USA
E-mail: kblee@rci.rutgers.edu

^g The Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, USA

Abstract

We have successfully designed and fabricated an integrated microfluidic platform, the hESC-µChip, 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.

Lab on a Chip

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

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

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