Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4419

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Preparing Embryonic Stem (ES) Cells for Aggregation

Andras Nagy, Marina Gertsenstein, Kristina Vintersten and Richard Behringer

This protocol was adapted from "Production of Chimeras," Chapter 11, in Manipulating the Mouse Embryo, 3rd edition, by Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

The first 15% of the full text of this article appears below.


INTRODUCTION

Ths protocol describes the preparation of ES cells, which can then be aggregated with diploid or tetraploid embryos to produce aggregation chimeras.


MATERIALS

Reagents

Aggregation plates, equilibrated (see Preparing the Aggregation Plate)

Embryos, zona-free (see Removal of Zona Pellucida)

ES cells, subconfluent, grown on 6-cm tissue culture dish with mouse embryo fibroblast (MEF) feeder layer (see De Novo Isolation of Embryonic Stem (ES) Cell Lines from Blastocysts)

recipe ES cell culture . . . [Full Text of this Article]

Equipment


METHOD


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