Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4428

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Immunosurgery: Isolating the Inner Cell Mass (ICM) of Blastocysts

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

This protocol describes a method for isolating the ICM of blastocysts by selectively killing the outer trophectoderm (TE).


MATERIALS

Reagents

Agar (Difco Noble) or agarose (Sigma; Calbiochem)

Dulbecco’s modified Eagle’s medium (DMEM) or similar medium

Fetal bovine serum (FBS), heat-inactivated

Guinea pig serum

Purchased (e.g., Murex Diagnostics or GIBCO) as freeze-dried guinea pig serum or prepared from fresh guinea pig blood obtained by cardiac puncture (see Step 2).

HEPES-buffered DMEM (20 mM) plus 10% heat-inactivated FBS

recipe M2 medium

Mouse spleen or tissue culture cells (see Step 1)

. . . [Full Text of this Article]

Equipment


METHOD


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