title: An integrated microfluidic culture device for quantitative analysis of human embryonic stem cells.
abstract: We have successfully designed and fabricated an integrated microfluidic platform, the hESC-microChip, 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.
authors.affiliation: Department of Molecular & Medical Pharmacology, University of California, Los Angeles, CA 90095, USA.
authors.forename: Ken-Ichiro
authors.initials: K
authors.lastname: Kamei
authors.forename: Shuling
authors.initials: S
authors.lastname: Guo
authors.forename: Zeta Tak For
authors.initials: ZT
authors.lastname: Yu
authors.forename: Hiroko
authors.initials: H
authors.lastname: Takahashi
authors.forename: Eric
authors.initials: E
authors.lastname: Gschweng
authors.forename: Carol
authors.initials: C
authors.lastname: Suh
authors.forename: Xiaopu
authors.initials: X
authors.lastname: Wang
authors.forename: Jinghua
authors.initials: J
authors.lastname: Tang
authors.forename: Jami
authors.initials: J
authors.lastname: McLaughlin
authors.forename: Owen N
authors.initials: ON
authors.lastname: Witte
authors.forename: Ki-Bum
authors.initials: KB
authors.lastname: Lee
authors.forename: Hsian-Rong
authors.initials: HR
authors.lastname: Tseng
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D048708:Cell Growth Processes; D049109:Cell Proliferation; D053595:Embryonic Stem Cells / Q000166:cytology*; D004867:Equipment Design; D006801:Humans; D007150:Immunohistochemistry; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D013997:Time Factors