title: Viability of zebrafish (Danio rerio) ovarian follicles after vitrification in a metal container.
abstract: Cryopreservation of ovarian tissue has been studied for female germline preservation of farm animals and endangered mammalian species. However, there are relatively few reports on cryopreservation of fish ovarian tissue and especially using vitrification approach. Previous studies of our group has shown that the use of a metal container for the cryopreservation of bovine ovarian fragments results in good primordial and primary follicle morphological integrity after vitrification. The aim of this study was to assess the viability and in vitro development of zebrafish follicles after vitrification of fragmented or whole ovaries using the same metal container. In Experiment 1, we tested the follicular viability of five developmental stages following vitrification in four vitrification solutions using fluorescein diacetate and propidium iodide fluorescent probes. These results showed that the highest viability rates were obtained with immature follicles (Stage I) and VS1 (1.5 M methanol + 4.5 M propylene glycol). In Experiment 2, we used VS1 to vitrify different types of ovarian tissue (fragments or whole ovaries) in two different carriers (plastic cryotube or metal container). In this experiment, Stage I follicle survival was assessed following vitrification by vital staining after 24 h in vitro culture. Follicular morphology was analyzed by light microscopy after vitrification. Data showed that the immature follicles morphology was well preserved after cryopreservation. Follicular survival rate was higher (P < 0.05) in vitrified fragments, when compared to whole ovaries. There were no significant differences in follicular survival and growth when the two vitrification devices were compared.
authors.affiliation: Aquam Research Group, Department of Animal Science, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil. Electronic address: lismarx@gmail.com.
authors.forename: Lis S
authors.initials: LS
authors.lastname: Marques
authors.affiliation: Department of Morphological Sciences, ICBS, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
authors.forename: Adriana
authors.initials: A
authors.lastname: Bos-Mikich
authors.affiliation: Aquaculture Graduate Program, Nilton Lins University, Manaus, AM, Brazil.
authors.forename: Leandro C
authors.initials: LC
authors.lastname: Godoy
authors.affiliation: Aquam Research Group, Department of Animal Science, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Silva
authors.affiliation: Aquam Research Group, Department of Animal Science, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
authors.forename: Daniel
authors.initials: D
authors.lastname: Maschio
authors.affiliation: Faculty of Science and Technology, Bournemouth University, Dorset, UK.
authors.forename: Tiantian
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Aquam Research Group, Department of Animal Science, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
authors.forename: Danilo P
authors.initials: DP
authors.lastname: Streit
mesh_terms: D000818:Animals; D002417:Cattle; D015925:Cryopreservation / Q000295:instrumentation* / Q000379:methods*; D005260:Female; D006801:Humans; D008670:Metals; D006080:Ovarian Follicle / Q000254:growth & development; D058989:Vitrification; D015027:Zebrafish
chemical_list: D008670:Metals
keywords: Cryopreservation; Female infertility; Fish; Follicle; Ovary