Abstract
A microfluidic human pluripotent stem cell (hPSC) array has been developed for robust and reproducible hPSC culture methods to assess chemically defined serum- and feeder-free culture conditions. This microfluidic platform, combined with image cytometry, enables the systematic analysis of multiple simultaneously detected marker expression in individual cells, for screening of various chemically defined media across hPSC lines, and the study of phenotypic responses.
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References
Giuliano KA et al (1997) High-content screening: a new approach to easing key bottlenecks in the drug discovery process. J Biomol Screen 2(4):249–259
Abraham VC, Taylor DL, Haskins JR (2004) High content screening applied to large-scale cell biology. Trends Biotechnol 22(1):15–22
Kamei K et al (2009) An integrated microfluidic culture device for quantitative analysis of human embryonic stem cells. Lab Chip 9(4):555–563
Kamei K et al (2010) Microfluidic image cytometry for quantitative single-cell profiling of human pluripotent stem cells in chemically defined conditions. Lab Chip 10(9):1113–1119
Morrison SJ, Spradling AC (2008) Stem cells and niches: mechanisms that promote stem cell maintenance throughout life. Cell 132(4):598–611
Wang L et al (2007) Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling. Blood 110(12):4111–4119
Ludwig TE et al (2006) Feeder-independent culture of human embryonic stem cells. Nat Methods 3(8):637–646
Ludwig TE et al (2006) Derivation of human embryonic stem cells in defined conditions. Nat Biotechnol 24(2):185–187
Yao S et al (2006) Long-term self-renewal and directed differentiation of human embryonic stem cells in chemically defined conditions. Proc Natl Acad Sci U S A 103(18):6907–6912
Carpenter AE et al (2006) Cell Profiler: image analysis software for identifying and quantifying cell phenotypes. Genome Biol 7:R100
Acknowledgements
This work was supported by the Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research at the Institute of Molecular Medicine at University of California, Los Angeles. Funding was also provided by the Terumo Life Science Foundation. The WPI-iCeMS is supported by the World Premier International Research Centre Initiative (WPI), the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.
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Mashimo, Y., Kamei, Ki. (2015). Microfluidic Image Cytometry for Single-Cell Phenotyping of Human Pluripotent Stem Cells. In: Singh, A., Chandrasekaran, A. (eds) Single Cell Protein Analysis. Methods in Molecular Biology, vol 1346. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2987-0_7
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DOI: https://doi.org/10.1007/978-1-4939-2987-0_7
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2986-3
Online ISBN: 978-1-4939-2987-0
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