한빛사 논문
Ken-ichiro Kamei‡ab, Shuling Guo‡cd, Zeta Tak For Yu‡abe, Hiroko Takahashiab, Eric Gschwengcd, Carol Suhab, Xiaopu Wangab, Jinghua Tangd, Jami McLaughlinc, Owen N. Witte*acdg, Ki-Bum Lee*abf and Hsian-Rong Tseng*ab
aDepartment of Molecular & Medical Pharmacology, University of California, Los Angeles, CA 90095, USA.
bCrump Institute for Molecular Imaging, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA
cDepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA 90095, USA
dThe Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, CA 90095, USA.
eDepartment of Mechanical and Aerospace Engineering, University of California, Los Angeles, CA 90095, USA
fDepartment 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, NJ 08854, USA.
gThe Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA
†Electronic supplementary information (ESI) available: Fabrication of hESC-µChips, generation of genetically modified hESCs (i.e., HSF1-LG, HSF1-OCT4-EGFP and H1-OCT4-EGFP), microscopy settings and induction of hESC differentiation. See DOI: 10.1039/b809105f
‡These three authors contributed equally to this work.
*Corresponding author.
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.
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