Microfluidic image cytometry for quantitative single-cell profiling of human pluripotent stem cells in chemically defined conditions

Ken-ichiro Kamei; Minori Ohashi; Eric Gschweng; Quinn Ho; Jane Suh; Jinghua Tang; Zeta Tak For Yu; Amander T. Clark; April D. Pyle; Michael A. Teitell; Ki-Bum. Lee; Owen N. Witte; Hsian-Rong Tseng

doi:10.1039/B922884E

Microfluidic image cytometry for quantitative single-cell profiling of human pluripotent stem cells in chemically defined conditions†

Ken-ichiro Kamei,‡*^abcde Minori Ohashi,‡^abcde Eric Gschweng,^ef Quinn Ho,^abcdeg Jane Suh,^abcde Jinghua Tang,^e Zeta Tak For Yu,^abcde Amander T. Clark,^eh April D. Pyle,^ef Michael A. Teitell,^ei Ki-Bum. Lee,^j Owen N. Witte^acefk and Hsian-Rong Tseng*^abcde

Author affiliations

* Corresponding authors

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

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

^c California NanoSystems Institute, University of California, Los Angeles, CA, USA

^d Institute for Molecular Medicine, University of California, Los Angeles, CA, USA

^e Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, CA, USA

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

^g Department of Biological Chemistry, University of California, Los Angeles, CA, USA

^h Department of Molecular Cell and Developmental Biology, Los Angeles, CA, USA

ⁱ Department of Pathology and Laboratory Medicine, Los Angeles, CA, USA

^j Department of Chemistry & Chemical Biology, Institute for Advanced Materials, Devices and Nanotechnology, Rutgers Stem Cell Research Center, Rutgers, The State University of New Jersey, Piscataway, NJ, USA

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

Abstract

Microfluidic image cytometry (MIC) has been developed to study phenotypes of various hPSC lines by screening several chemically defined serum/feeder-free conditions. A chemically defined hPSC culture was established using 20 ng mL⁻¹ of bFGF on 20 μg mL⁻¹ of Matrigel to grow hPSCs over a week in an undifferentiated state. Following hPSC culture, we conducted quantitative MIC to perform a single cell profiling of simultaneously detected protein expression (OCT4 and SSEA1). Using clustering analysis, we were able to systematically compare the characteristics of various hPSC lines in different conditions.

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Microfluidic image cytometry for quantitative single-cell profiling of human pluripotent stem cells in chemically defined conditions†

Abstract

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Microfluidic image cytometry for quantitative single-cell profiling of human pluripotent stem cells in chemically defined conditions

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