title: Maximizing Clonal Embryonic Stem Cell Derivation by ERK Pathway Inhibition.
abstract: Since the development of inhibitor-based defined culture conditions (known as "2i"), multiple clonal embryonic stem cell (ESC) lines can be readily derived from single cells isolated directly from mouse embryos. In addition to providing an efficient means to generate ES cells from compound transgenic or murine disease models on any genetic background, this technology can be used to investigate the process of ESC derivation at both a functional and molecular level. Here, we provide details of the procedure for both maximizing the number of cells in the donor tissue and subsequent effective derivation of multiple clonal ES cell lines.
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK. jn270@cam.ac.uk.|Department of Physiology, Development and Neuroscience, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK. jn270@cam.ac.uk.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Nichols
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Boroviak
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D002999:Clone Cells; D046149:Embryo Culture Techniques / Q000379:methods*; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*
chemical_list: D047428:Protein Kinase Inhibitors
keywords: 2i; Blastocyst; ERK/MAPK inhibition; Embryonic stem cells; Epiblast; Gsk3 inhibition; Pluripotency