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  • Generation of Mammalian offspring by haploid embryonic stem cells microinjection.

Generation of Mammalian offspring by haploid embryonic stem cells microinjection.

Current protocols in stem cell biology (2014-11-05)
Ling Shuai, Wei Li, Haifeng Wan, Xiao-Yang Zhao, Liu Wang, Qi Zhou
ABSTRACT

In this unit we introduce the derivation and genetic modification of mouse haploid embryonic stem (ES) cells. We detail how to produce haploid embryos and the subsequent ES derivation and cell culture. We further introduce readers to the intracytoplasmic injection processes of two types of haploid ES cells [androgenetic haploid ES (ahES) and parthenogenetic ES (phES)], both of which possess potential to produce fertile progenies by microinjection. This unit will be interesting to researchers who focus on recessive screens and transgenic animal model production with haploid stem cells.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Bovine Serum Albumin, powder, BioXtra
Sigma-Aldrich
(±)-Verapamil hydrochloride, ≥99% (titration), powder
Sigma-Aldrich
Mitomycin C from Streptomyces caespitosus, powder, contains NaCl as solubilizer
Sigma-Aldrich
Chorionic gonadotropin human, lyophilized powder, from human pregnancy urine
Sigma-Aldrich
ESGRO® Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.