Isolation and in vitro Culture of Mouse Oocytes.

Autor:	Greaney J; The Christopher Chen Oocyte Biology Research Laboratory, University of Queensland Centre for Clinical Research, The University of Queensland, Brisbane, Australia., Subramanian GN; The Christopher Chen Oocyte Biology Research Laboratory, University of Queensland Centre for Clinical Research, The University of Queensland, Brisbane, Australia., Ye Y; The Christopher Chen Oocyte Biology Research Laboratory, University of Queensland Centre for Clinical Research, The University of Queensland, Brisbane, Australia., Homer H; The Christopher Chen Oocyte Biology Research Laboratory, University of Queensland Centre for Clinical Research, The University of Queensland, Brisbane, Australia.
Jazyk:	angličtina
Zdroj:	Bio-protocol [Bio Protoc] 2021 Aug 05; Vol. 11 (15), pp. e4104. Date of Electronic Publication: 2021 Aug 05 (Print Publication: 2021).
DOI:	10.21769/BioProtoc.4104
Abstrakt:	Females are endowed at birth with a fixed reserve of oocytes, which declines both in quantity and quality with advancing age. Understanding the molecular mechanisms regulating oocyte quality is crucial for improving the chances of pregnancy success in fertility clinics. In vitro culture systems enable researchers to analyse important molecular and genetic regulators of oocyte maturation and fertilisation. Here, we describe in detail a highly reproducible technique for the isolation and culture of fully grown mouse oocytes. We include the considerations and precautionary measures required for minimising the detrimental effects of in vitro culture conditions. This technique forms the starting point for a wide range of experimental approaches such as post-transcriptional gene silencing, immunocytochemistry, Western blotting, high-resolution 4D time-lapse imaging, and in vitro fertilization, which are instrumental in dissecting the molecular determinants of oocyte quality. Hence, this protocol serves as a useful, practical guide for any oocyte researcher beginning experiments aimed at investigating important oocyte molecular factors. Graphic abstract: A step-by-step protocol for the isolation and in vitro culture of oocytes from mice. Competing Interests: Competing interests H.A.H. is a co-founder, shareholder, and advisor of Jumpstart Fertility Inc., which was founded to develop research into NAD+-dependent pathways involved in female fertility. H.A.H. is a co-inventor on the patent application ‘Methods for increasing fertility’ (application number 2017903013). All remaining authors declare no competing interests. (Copyright © 2021 The Authors; exclusive licensee Bio-protocol LLC.)
Databáze:	MEDLINE
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