{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T15:01:44Z","timestamp":1790694104062,"version":"4.1.0"},"reference-count":27,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,8,25]],"date-time":"2021-08-25T00:00:00Z","timestamp":1629849600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,8,25]],"date-time":"2021-08-25T00:00:00Z","timestamp":1629849600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100004189","name":"Max-Planck-Gesellschaft","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004189","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Max Planck Institute for Biology of Ageing"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Over the last decade, the African turquoise killifish,\n                    <jats:italic>Nothobranchius furzeri,<\/jats:italic>\n                    has emerged as an important model system for the study of vertebrate biology and ageing. Propagation of laboratory inbred strains of\n                    <jats:italic>Nothobranchius furzeri<\/jats:italic>\n                    , 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 ~\u200925% and peaks of ~\u200955% fertilization. This technical advance will greatly facilitate the use of\n                    <jats:italic>N. furzeri<\/jats:italic>\n                    as a model organism.\n                  <\/jats:p>","DOI":"10.1038\/s41598-021-96383-8","type":"journal-article","created":{"date-parts":[[2021,8,25]],"date-time":"2021-08-25T06:46:44Z","timestamp":1629874004000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Sperm cryopreservation and in vitro fertilization techniques for the African turquoise killifish Nothobranchius furzeri"],"prefix":"10.1038","volume":"11","author":[{"given":"Luca","family":"Dolfi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tsz Kin","family":"Suen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberto","family":"Ripa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam","family":"Antebi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,8,25]]},"reference":[{"key":"96383_CR1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1474-9726.2005.00165.x","author":"T Genade","year":"2005","unstructured":"Genade, T. et al. Annual fishes of the genus Nothobarnchius as a model system for aging research. Aging Cell https:\/\/doi.org\/10.1111\/j.1474-9726.2005.00165.x (2005).","journal-title":"Aging Cell"},{"key":"96383_CR2","doi-asserted-by":"publisher","DOI":"10.1111\/brv.12183","author":"A Cellerino","year":"2015","unstructured":"Cellerino, A., Valenzano, D. R. & Reichard, M. From the bush to the bench: The annual Nothobranchius fishes as a new model system in biology. Biol. Rev. https:\/\/doi.org\/10.1111\/brv.12183 (2015).","journal-title":"Biol. Rev."},{"key":"96383_CR3","doi-asserted-by":"publisher","first-page":"1013","DOI":"10.1016\/j.cell.2015.01.038","volume":"160","author":"I Harel","year":"2015","unstructured":"Harel, I. et al. A platform for rapid exploration of aging and diseases in a naturally short-lived vertebrate. Cell 160, 1013\u20131026 (2015).","journal-title":"Cell"},{"key":"96383_CR4","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.ecolsys.31.1.139","author":"PW Hedrick","year":"2000","unstructured":"Hedrick, P. W. Inbreeding depression in conservation biology. Annu. Rev. Ecol. Syst. https:\/\/doi.org\/10.1146\/annurev.ecolsys.31.1.139 (2000).","journal-title":"Annu. Rev. Ecol. Syst."},{"key":"96383_CR5","doi-asserted-by":"publisher","DOI":"10.1007\/s10592-007-9405-0","author":"LI Wright","year":"2008","unstructured":"Wright, L. I., Tregenza, T. & Hosken, D. J. Inbreeding, inbreeding depression and extinction. Conserv. Genet. https:\/\/doi.org\/10.1007\/s10592-007-9405-0 (2008).","journal-title":"Conserv. Genet."},{"key":"96383_CR6","doi-asserted-by":"publisher","DOI":"10.1089\/zeb.2018.1631","author":"M Api","year":"2018","unstructured":"Api, M., Notarstefano, V., Olivotto, I., Cellerino, A. & Carnevali, O. Breeders age affects reproductive success in Nothobranchius furzeri. Zebrafish https:\/\/doi.org\/10.1089\/zeb.2018.1631 (2018).","journal-title":"Zebrafish"},{"key":"96383_CR7","doi-asserted-by":"publisher","DOI":"10.3791\/57073","author":"J Dodzian","year":"2018","unstructured":"Dodzian, J., Kean, S., Seidel, J. & Valenzano, D. R. A protocol for laboratory housing of Turquoise killifish (Nothobranchius furzeri). J. Vis. Exp. https:\/\/doi.org\/10.3791\/57073 (2018).","journal-title":"J. Vis. Exp."},{"key":"96383_CR8","doi-asserted-by":"publisher","DOI":"10.1016\/j.exger.2020.111073","volume":"140","author":"J \u017d\u00e1k","year":"2020","unstructured":"\u017d\u00e1k, J. & Reichard, M. Fluctuating temperatures extend median lifespan, improve reproduction and reduce growth in turquoise killifish. Exp. Gerontol. 140, 111073 (2020).","journal-title":"Exp. Gerontol."},{"key":"96383_CR9","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1534\/g3.111.001271","volume":"1","author":"DR Valenzano","year":"2011","unstructured":"Valenzano, D. R., Sharp, S., Brunet, A. & Andrews, B. J. Transposon-mediated transgenesis in the short-lived African killifish Nothobranchius furzeri, a vertebrate model for aging. G3 Genes Genomes Genetics 1, 531\u2013538 (2011).","journal-title":"G3 Genes Genomes Genetics"},{"key":"96383_CR10","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.103","author":"I Harel","year":"2016","unstructured":"Harel, I., Valenzano, D. R. & Brunet, A. Efficient genome engineering approaches for the short-lived African turquoise killifish. Nat. Protoc. https:\/\/doi.org\/10.1038\/nprot.2016.103 (2016).","journal-title":"Nat. Protoc."},{"key":"96383_CR11","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1804590115","author":"ALT Romney","year":"2018","unstructured":"Romney, A. L. T., Davis, E. M., Corona, M. M., Wagner, J. T. & Podrabsky, J. E. Temperature-dependent Vitamin D signaling regulates developmental trajectory associated with diapause in an annual killifish. Proc. Natl. Acad. Sci. 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Mechanisms of fertilization in fishes. Integr. Comp. Biol. https:\/\/doi.org\/10.1093\/icb\/21.2.359 (1981).","journal-title":"Integr. Comp. Biol."},{"key":"96383_CR27","doi-asserted-by":"publisher","DOI":"10.2307\/1541457","author":"FJ Longo","year":"1982","unstructured":"Longo, F. J., So, F. & Schuetz, A. W. Meiotic maturation and the cortical granule reaction in starfish eggs. Biol. Bull. https:\/\/doi.org\/10.2307\/1541457 (1982).","journal-title":"Biol. 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