Abstract
Experimental manipulation of hESCs has been hampered by their fragility and susceptibility to apoptosis when dissociated into single cells. The OxF lines are particularly robust and may be successfully passaged as single cells, with the inclusion of ROCK inhibitor in the medium. The protocols here describe the enzymatic dissociation of hESCs into a single-cell suspension and the plating of these cells onto either feeder cells or a protein-coated surface.
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References
Brook FA et al (2010) Derivation and characterisation of the human embryonic stem cell line, OxF1. In Vitro Cell Dev Biol Anim 46: 173–177
Watanabe K et al (2007) A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nature Biotech 25: 681–686
Brook FA (2006) Procedures for deriving ES cell lines from the mouse. In: Notarianni E, Evans MJ (eds) Embryonic stem cells. A practical approach. Oxford University Press, Oxford, pp 7–40
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Brook, F.A. (2012). Single-Cell Enzymatic Dissociation of hESC Lines OxF1–OxF4 and Culture in Feeder-Free Conditions. In: Turksen, K. (eds) Human Embryonic Stem Cells Handbook. Methods in Molecular Biology, vol 873. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-794-1_12
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DOI: https://doi.org/10.1007/978-1-61779-794-1_12
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-793-4
Online ISBN: 978-1-61779-794-1
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