Microfluidic image cytometry for quantitative single-cell profiling of human pluripotent stem cells in chemically defined conditions.
Publication Year:
2010
PubMed ID:
20390128
Funding Grants:
- Mitochondrial Metabolism in hESC and hiPSC Differentiation, Reprogramming, and Cancer
- Role of Mitochondria in Self-Renewal Versus Differentiation of Human Embryonic Stem Cells
- Generation of Pluripotent Cell Lines from Human Embryos
- Microfluidic Platform for Screening Chemically Defined Conditions that Facilitate Clonal Expansion of Human Pluripotent Stem Cells
Scientific 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 microg 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.