Chapter 23 – Approaches for Derivation and Maintenance of Human ES Cells: Detailed Procedures and Alternatives

Success of hESC derivation strongly relies on mastering different techniques and close attention to details. We describe several techniques and approaches which allowed us to derive over 30 hESC lines including derivation under feeder-free conditions and from a single blastomere. The Chapter provides advice and protocols for different aspects of hESC culture including laboratory setup, quality of reagents, early derivation steps, adaptation to trypsin, freezing-thawing with high yield of surviving cells, etc.

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