Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4399

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Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomycin C

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


INTRODUCTION

This protocol describes the preparation of feeder cell layers from MEF cells or from the STO mouse fibroblast cell line. The cells are rendered mitotically inactive by treatment with mitomycin C. The feeder layers can then be used to maintain embryonic stem (ES) cells in the undifferentiated state. For an alternative protocol, see Preparing Feeder Cell Layers from STO or Mouse Embryo . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


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