通过八细胞期胚胎注射产生兔嵌合体。

Generation of Rabbit Chimeras by Eight-Cell Stage Embryo Injection.

作者信息

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

机构信息

Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center, Ann Arbor, MI, USA.

Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University, Harbin, China.

出版信息

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

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

PMID:36995679

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.

摘要

将多能干细胞（PSC）注射到囊胚期胚胎中是一种通过嵌合贡献来评估多能性的广泛使用的方法。它通常用于生产转基因小鼠。然而，将PSC注射到兔囊胚期胚胎中具有挑战性。在此阶段，体内发育的兔囊胚具有一层厚厚的粘蛋白层，这对显微注射具有抑制作用，而缺乏这种粘蛋白层的体外发育的兔囊胚在胚胎移植后往往无法着床。在本章中，我们描述了一种通过无粘蛋白八细胞期胚胎注射程序生产兔嵌合体的详细方案。