Sperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri

Abstract Over the last decade, the African turquoise killifish, Nothobranchius furzeri, has emerged as an important model system for the study of vertebrate biology and ageing. Propagation of laboratory inbred strains of Nothobranchius furzeri, such as GRZ, however, can pose challenges due to the sh...

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Autores principales:	Luca Dolfi, Tsz Kin Suen, Roberto Ripa, Adam Antebi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/bf414b8d784a4b4084d5f45b26195222
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spelling	oai:doaj.org-article:bf414b8d784a4b4084d5f45b261952222021-12-02T18:53:14ZSperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri10.1038/s41598-021-96383-82045-2322https://doaj.org/article/bf414b8d784a4b4084d5f45b261952222021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-96383-8https://doaj.org/toc/2045-2322Abstract Over the last decade, the African turquoise killifish, Nothobranchius furzeri, has emerged as an important model system for the study of vertebrate biology and ageing. Propagation of laboratory inbred strains of Nothobranchius furzeri, such as GRZ, however, can pose challenges due to the short window of fertility, the efforts and space requirements involved in continuous strain maintenance, and the risks of further inbreeding. The current method for long term strain preservation relies on arrest of embryos in diapause. To create an alternative for long term maintenance, we developed a robust protocol to cryopreserve and revive sperm for in vitro fertilization (IVF). We tested a variety of extender and activator buffers for sperm IVF, as well as cryoprotectants to achieve practical long-term storage and fertilization conditions tailored to this species. Our protocol enabled sperm to be preserved in a cryogenic condition for months and to be revived with an average of 40% viability upon thawing. Thawed sperm were able to fertilize nearly the same number of eggs as natural fertilization, with an average of ~ 25% and peaks of ~ 55% fertilization. This technical advance will greatly facilitate the use of N. furzeri as a model organism.Luca DolfiTsz Kin SuenRoberto RipaAdam AntebiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Luca Dolfi Tsz Kin Suen Roberto Ripa Adam Antebi Sperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri
description	Abstract Over the last decade, the African turquoise killifish, Nothobranchius furzeri, has emerged as an important model system for the study of vertebrate biology and ageing. Propagation of laboratory inbred strains of Nothobranchius furzeri, such as GRZ, however, can pose challenges due to the short window of fertility, the efforts and space requirements involved in continuous strain maintenance, and the risks of further inbreeding. The current method for long term strain preservation relies on arrest of embryos in diapause. To create an alternative for long term maintenance, we developed a robust protocol to cryopreserve and revive sperm for in vitro fertilization (IVF). We tested a variety of extender and activator buffers for sperm IVF, as well as cryoprotectants to achieve practical long-term storage and fertilization conditions tailored to this species. Our protocol enabled sperm to be preserved in a cryogenic condition for months and to be revived with an average of 40% viability upon thawing. Thawed sperm were able to fertilize nearly the same number of eggs as natural fertilization, with an average of ~ 25% and peaks of ~ 55% fertilization. This technical advance will greatly facilitate the use of N. furzeri as a model organism.
format	article
author	Luca Dolfi Tsz Kin Suen Roberto Ripa Adam Antebi
author_facet	Luca Dolfi Tsz Kin Suen Roberto Ripa Adam Antebi
author_sort	Luca Dolfi
title	Sperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri
title_short	Sperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri
title_full	Sperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri
title_fullStr	Sperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri
title_full_unstemmed	Sperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri
title_sort	sperm cryopreservation and in vitro fertilization techniques for the african turquoise killifish nothobranchius furzeri
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/bf414b8d784a4b4084d5f45b26195222
work_keys_str_mv	AT lucadolfi spermcryopreservationandinvitrofertilizationtechniquesfortheafricanturquoisekillifishnothobranchiusfurzeri AT tszkinsuen spermcryopreservationandinvitrofertilizationtechniquesfortheafricanturquoisekillifishnothobranchiusfurzeri AT robertoripa spermcryopreservationandinvitrofertilizationtechniquesfortheafricanturquoisekillifishnothobranchiusfurzeri AT adamantebi spermcryopreservationandinvitrofertilizationtechniquesfortheafricanturquoisekillifishnothobranchiusfurzeri
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Sperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri

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