Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4407

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Derivation of Trophoblast Stem (TS) Cell Lines from Blastocysts

Janet Rossant

This protocol was adapted from "Isolation and Culture of Blastocyst-Derived Stem Cell Lines", Chapter 8 (Protocol 9, from the laboratory of Janet Rossant), 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 100 words of the full text of this article appear below.


INTRODUCTION

This protocol decribes derivation of TS cell lines from 3.5-days post coitum (dpc) mouse blastocysts. The procedure is similar to the derivation of embryonic stem (ES) cell lines. However, the success rate is considerably higher, and less expertise is required to recognize pluripotent TS cell colonies.


MATERIALS

Reagents

Expanded blastocysts (3.5 dpc) (see Collecting Blastocysts)

recipe Feeder-conditioned medium (feeder-CM)

recipe FGF4 stock solution (1000X, 25 µg/ml)

Heparin stock solution (1000X, 1.0 mg/ml) (Sigma, 10,000 units)

recipeResuspend in PBS and store at -80ºC. The stock can be also prepared as 10,000X (10 mg/ml) solution.

Mouse embryo fibroblast (MEF) feeder layer (see Preparing Feeder Cell . . . [Full Text of this Article]

Equipment


METHOD


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