Universität zu Köln

Wigger, Magdalena ORCID: 0000-0002-5777-7967, Troder, Simon E. and Zevnik, Branko ORCID: 0000-0001-7845-9522 (2021). A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm. PLoS One, 16 (10). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

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Abstract

The advent of genome editing tools like CRISPR/Cas has substantially increased the number of genetically engineered mouse models in recent years. In support of refinement and reduction, sperm cryopreservation is advantageous compared to embryo freezing for archiving and distribution of such mouse models. The in vitro fertilization using cryopreserved sperm from the most widely used C57BL/6 strain has become highly efficient in recent years due to several improvements of the procedure. However, purchase of the necessary media for routine application of the current protocol poses a constant burden on budgetary constraints. In-house media preparation, instead, is complex and requires quality control of each batch. Here, we describe a cost-effective and easily adaptable approach for in vitro fertilization using cryopreserved C57BU6 sperm. This is mainly achieved by modification of an affordable commercial fertilization medium and a step-by-step description of all other necessary reagents. Large-scale comparison of fertilization rates from independent lines of genetically engineered C57BL/6 mice upon cryopreservation and in vitro fertilization with our approach demonstrated equal or significantly superior fertilization rates to current protocols. Our novel SEcuRe (Simple Economical set-up for Rederivation) method provides an affordable, easily adaptable and harmonized protocol for highly efficient rederivation using cryopreserved C57BL/6 sperm for a broad application of colony management in the sense of the 3Rs.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Wigger, Magdalena UNSPECIFIED orcid.org/0000-0002-5777-7967 UNSPECIFIED Troder, Simon E. UNSPECIFIED UNSPECIFIED UNSPECIFIED Zevnik, Branko UNSPECIFIED orcid.org/0000-0001-7845-9522 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-567279
DOI:	10.1371/journal.pone.0259202
Journal or Publication Title:	PLoS One
Volume:	16
Number:	10
Date:	2021
Publisher:	PUBLIC LIBRARY SCIENCE
Place of Publication:	SAN FRANCISCO
ISSN:	1932-6203
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language METHYL-BETA-CYCLODEXTRIN; HUMAN TUBAL FLUID; MALE-MICE; SPERMATOZOA; CULTURE; EMBRYOS; CHOLESTEROL; FERTILITY; SURVIVAL Multiple languages Multidisciplinary Sciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/56727