Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4402

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Freezing and Thawing of Embryonic Stem (ES) Cells Using Cryovials

Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer

This protocol was adapted from "Isolation and Culture of Blastocyst-Derived Stem Cell Lines," Chapter 8 of 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 100 words of the full text of this article appear below.


INTRODUCTION

This protocol describes a method for freezing and thawing ES cells using cryovials. It is important to freeze ES cell stocks as soon as possible to reduce the time that they are in culture. A careful record should be kept of the number of times cells are passaged and the location of the cryovials.


MATERIALS

Reagents

caution Dimethyl sulfoxide (DMSO) (e.g., Sigma)

ES cells (subconfluent plate ready to be frozen) (see De Novo Isolation of Embryonic Stem (ES) Cell Lines from Blastocysts)

ES cells, frozen in cryovial (for thawing procedure) (see De Novo Isolation of Embryonic Stem (ES) Cell Lines from Blastocysts. . . [Full Text of this Article]

Equipment


METHOD


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