Generation of Rabbit Chimeras by Eight-Cell Stage Embryo Injection

Dongshan Yang; Jun Song; Jie Xu; Jifeng Zhang; Y Eugene Chen

doi:10.1007/978-1-0716-2990-1_18

Generation of Rabbit Chimeras by Eight-Cell Stage Embryo Injection

Methods Mol Biol. 2023:2631:381-391. doi: 10.1007/978-1-0716-2990-1_18.

Authors

Dongshan Yang¹, Jun Song^{2

3}, Jie Xu², Jifeng Zhang², Y Eugene Chen⁴

Affiliations

¹ Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center, Ann Arbor, MI, USA. doyang@med.umich.edu.
² Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center, Ann Arbor, MI, USA.
³ Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University, Harbin, China.
⁴ Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center, Ann Arbor, MI, USA. echenum@umich.edu.

PMID: 36995679
DOI: 10.1007/978-1-0716-2990-1_18

Abstract

Pluripotent stem cell (PSC) injection to the blastocyst stage embryos is a widely used method to evaluate the pluripotency through chimeric contribution. It is routinely used to produce transgenic mice. However, PSC injection to the blastocyst stage embryos in rabbits is challenging. At this stage, the in vivo developed rabbit blastocysts possess a thick mucin layer that is inhibitory for microinjection, whereas in vitro developed rabbit blastocysts that lack such mucin layer often fail to implant after embryo transfer. In this chapter, we describe a detailed protocol of rabbit chimera production through mucin-free eight-cell stage embryo injection procedure.

Keywords: Chimera; Eight-cell stage embryo injection; Pluripotent stem cells; Rabbit.

MeSH terms

Animals
Blastocyst
Embryo Transfer / methods
Embryo, Mammalian*
Mice
Microinjections
Pluripotent Stem Cells*
Rabbits