Journal article

An efficient method for the derivation of mouse embryonic stem cells

Vitezslav Bryja, Sonia Bonilla, Lukas Cajanek, Clare L Parish, Catherine M Schwartz, Yongquan Luo, Mahendra S Rao, Ernest Arenas

Stem Cells | ALPHAMED PRESS | Published : 2006

Abstract

Mouse embryonic stem cells (mESCs) represent a unique tool for many researchers; however, the process of ESC derivation is often very inefficient and requires high specialization, training, and expertise. To circumvent these limitations, we aimed to develop a simple and efficient protocol based on the use of commercially available products. Here, we present an optimized protocol that we successfully applied to derive ESCs from several knockout mouse strains (Wnt-1, Wnt-5a, Lrp6, and parkin) with 50%-75% efficiency. The methodology is based on the use of mouse embryonic fibroblast feeders, knockout serum replacement (SR), and minimal handling of the blastocyst. In this protocol, all centrifug..

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