A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm

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 mo...

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Autores principales:	Magdalena Wigger, Simon E. Tröder, Branko Zevnik
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/35a808a3691443ad95f35a89009f2236
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spelling	oai:doaj.org-article:35a808a3691443ad95f35a89009f22362021-11-04T06:49:33ZA simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm1932-6203https://doaj.org/article/35a808a3691443ad95f35a89009f22362021-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8553151/?tool=EBIhttps://doaj.org/toc/1932-6203The 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 C57BL/6 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.Magdalena WiggerSimon E. TröderBranko ZevnikPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Magdalena Wigger Simon E. Tröder Branko Zevnik A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm
description	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 C57BL/6 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.
format	article
author	Magdalena Wigger Simon E. Tröder Branko Zevnik
author_facet	Magdalena Wigger Simon E. Tröder Branko Zevnik
author_sort	Magdalena Wigger
title	A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm
title_short	A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm
title_full	A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm
title_fullStr	A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm
title_full_unstemmed	A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm
title_sort	simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/35a808a3691443ad95f35a89009f2236
work_keys_str_mv	AT magdalenawigger asimpleandeconomicprotocolforefficientinvitrofertilizationusingcryopreservedmousesperm AT simonetroder asimpleandeconomicprotocolforefficientinvitrofertilizationusingcryopreservedmousesperm AT brankozevnik asimpleandeconomicprotocolforefficientinvitrofertilizationusingcryopreservedmousesperm AT magdalenawigger simpleandeconomicprotocolforefficientinvitrofertilizationusingcryopreservedmousesperm AT simonetroder simpleandeconomicprotocolforefficientinvitrofertilizationusingcryopreservedmousesperm AT brankozevnik simpleandeconomicprotocolforefficientinvitrofertilizationusingcryopreservedmousesperm
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A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm

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