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Abstract

Developmental Biology

Application of Mouse Parthenogenetic Haploid Embryonic Stem Cells as a Substitute of Sperm

Published: November 19th, 2020

DOI:

10.3791/61999

1Institute of Molecular Health Sciences, Swiss Federal Institute of Technology, ETH Zurich

Abstract

In organisms with sexual reproduction, germ cells are the source of totipotent cells that develop into new individuals. In mice, fertilization of an oocyte by a spermatozoon creates a totipotent zygote. Recently, several publications have reported that haploid embryonic stem cells (haESCs) can be a substitute for gametic genomes and contribute to embryos, which develop into mice. Here, we present a protocol to apply parthenogenetic haESCs as a substitute of sperm to construct embryos by intracytoplasmic injection into oocytes. This protocol consists of steps for preparing haESCs as sperm replacement, for injection of haESC chromosomes into oocytes, and for culture of semi-cloned embryos. The embryos can yield fertile semi-cloned mice after embryo transfer. Using haESCs as sperm replacement facilitates genome editing in the germline, studies of embryonic development, and investigation of genomic imprinting.

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Keywords Mouse Parthenogenetic Haploid Embryonic Stem Cells

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