Feasibility Studies on Vitrification of Intact Zebrafish (Brachydanio rerio) Embryos

Zhang, Tiantian; Rawson, David M.

doi:10.1006/cryo.1996.0001

Cited by 87 publications

(44 citation statements)

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“…The glass-forming tendencies of cryoprotectants and their lowest glass-forming concentrations during cooling observed in the absence of sucrose (3 M 1,3-BG, 4 M PG, 5 M DMSO, 5 M GC, 6 M EG and 10 M MeOH) were similar to the findings in previous studies in which cryoprotectant solutions were loaded in a 0.25-or 0.5-ml plastic straw (4 M PG, 5 M DMSO, 5 M GC, 6 to 6.5 M EG and ≥10 M MeOH) [21,22]. We confirmed that addition of sucrose to the cryoprotectant solutions decreased the concentrations of cryoprotectants in the solutions One-microliter solutions containing 1 to 10 M cryoprotectant at intervals of 1 M were tested.…”

Section: Discussionsupporting

confidence: 89%

Cryopreservation of Primordial Germ Cells by Rapid Cooling of Whole Zebrafish (Danio rerio) Embryos

Higaki

Mochizuki

Akashi

et al. 2010

Journal of Reproduction and Development

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Abstract. The feasibility of cryopreservation of zebrafish (Danio rerio) primordial germ cells (PGCs) by rapid cooling (i.e., vitrification) of dechorionated whole embryos at the 14-to 20-somite stage was investigated. Initially, we examined the glass-forming properties and embryo toxicities of six cryoprotectants: methanol (MeOH), ethylene glycol (EG), glycerol (GC), dimethyl sulfoxide (DMSO), propylene glycol (PG) and 1,3-butylene glycol (1,3-BG). According to the results of glass-forming and embryo toxicity tests, pretreatment solution (PS) containing 2 or 3 M cryoprotectant and vitrification solution (VS) containing 5 M cryoprotectant and 0.5 M sucrose were prepared using each cryoprotectant. Dechorionated embryos, the PGCs of which were visualized by injection of green fluorescence protein-nos1 3'UTR mRNA, were cooled rapidly by plunging into liquid nitrogen after serial exposure to PS and VS. All embryos cooled with MeOH, PG and 1,3-BG showed ice formation during cooling, and few embryos had live PGCs after warming. Most embryos cooled with GC did not show ice formation; however, few embryos had live PGCs. All embryos cooled with EG and most embryos cooled with DMSO had live PGCs when the embryos did not show ice formation during cooling. Based on the number of live PGCs in fresh embryos, the maximum survival rates of PGCs recovered from embryos cooled with EG and DMSO were estimated to be about 40 and 20%, respectively. The present study indicates that rapid cooling of dechorionated whole embryos, especially using EG-based solutions, could be utilized as a simple and promising tool for cryopreservation of PGCs.

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Section: Discussionsupporting

confidence: 89%

Cryopreservation of Primordial Germ Cells by Rapid Cooling of Whole Zebrafish (Danio rerio) Embryos

Higaki

Mochizuki

Akashi

et al. 2010

Journal of Reproduction and Development

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“…The DMSO is a widely used cryoprotectant during the cooling processes, both for the cryopreservation/ vitrification of semen and embryos and this cryoprotectant has as advantage the rapid permeation into the membrane (Hagedorn et al, 1997a(Hagedorn et al, , b, 1998Zhang & Rawson 1996). In our study the DMSO showed lower toxicity during incubation of the embryo for longer periods compared with ethylene glycol, being the cryoprotectant suggested for these conditions.…”

Section: Discussionsupporting

confidence: 47%

“…BSF is a compound of bovine blood proteins and may have been toxic to curimba embryos, which has not been observed for bovine embryos as it accounts for an increase in their growth (Lonergan et al, 1999). Zhang & Rawson (1996) measured the volume in the outer chorion to determine the cryoprotectant permeation of intact six-somite and 27h embryos. Importantly, after 30 minutes of exposure, all embryos shrank, except those which received EG GEL 0.5% treatment in our study.…”

Section: Discussionmentioning

confidence: 99%

Toxicity of cryoprotectants on Prochilodus lineatus (Valenciennes, 1837) (curimba) embryos in an experimental incubator (Characiformes: Prochilodontidae)

et al. 2014

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This paper investigated the effect of cryoprotectant substances on Prochilodus lineatus embryos in an experimental incubator. The prospective study applied combinations of polyvinyl alcohol, hydroxyethyl cellulose, gelatin and fetal bovine serum with dimethyl sulfoxide and ethylene glycol in a new experimental incubator. The morphology of embryos, larval viability and the efficiency of experimental incubators in maintaining the quality of embryos were evaluated. This study demonstrates the efficient association between hydroxyethylcellulose and dimethyl sulfoxide as greater viability (p<0.05) was found for embryos (72.9 ± 23.9%). It should also be noted the permeation of cryoprotectants in embryos through the changes found in chorion diameter, embryo diameter and embryo volume comparing the treatments versus control group (water) (p<0.05), this results can help in future cryopreservation protocols. Although the temperature and oxygenation differed between the usual and experimental incubators (p<0.05), the results showed a high fertilization rate (79.6 ± 13.2%) for experimental incubators (p<0.05) which is sufficient for the maintenance of embryos in a cryoprotective environment and effectively allows experimentation for long periods with cryoprotectant substances. Cryopreservation of fish embryos has not been accomplished yet and new approaches are required for understanding the permeability of teleost embryos, especially in Brazilian native species.Este trabalho avaliou o efeito de substâncias crioprotetoras sobre embriões de Prochilodus lineatus em uma incubadora experimental. O estudo aplicou combinações de álcool polivinílico, hidroxietilcelulose, gelatina e soro fetal bovino com dimetilosulfóxido e etilenoglicol em uma nova incubadora experimental. Foram avaliadas a morfologia dos embriões, a viabilidade larval e a eficiência das incubadoras experimentais na manutenção da qualidade dos embriões. Este estudo demonstra a associação eficiente entre hidroxietilcelulose e dimetilsulfóxido pela maior viabilidade (p<0,05) encontrada para os embriões (72,9 ± 23,9%). Deve-se notar também a permeação dos crioprotetores nos embriões através das alterações encontradas no diâmetro córion, diâmetro do embrião e no volume do embrião comparando os tratamentos ao grupo controle (água) (p<0,05), estes resultados podem ajudar em futuros protocolos de criopreservação. Embora a temperatura e a oxigenação diferiram entre as incubadoras comuns e as experimentais (p<0,05), os resultados mostraram elevada taxa de fertilização (79,6 ± 13,2%) para incubadoras experimentais (p<0,05), o que é suficiente para a manutenção de embriões em ambiente crioprotetor e permite efetivamente a experimentação por longos períodos com substâncias crioprotetoras. A criopreservação de embriões de peixes ainda não foi realizada e novas abordagens são necessárias para a compreensão da permeabilidade dos embriões de teleósteos, especialmente em espécies nativas brasileiras.

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“…In many of these systems, either dimethyl sulfoxide (McjSO) or propylene glycol (PG) are the least toxic of various cryoprotectants tested in the following embryos: rainbow trout Oncorhynchus mykiss, medaka Oryzias latipes, and pejerrey Odonthestes bonariensis (30); red drum Sciaenops ocellatus (23); rotifer Brachionus plicatilis (32); rosy barb Puntius conchonius (1); blue mussel Mytilus edulis (31); and zebrafish embryos (13,38). However, it was unknown what effect these two cryoprotectants might have when introduced intracellularly for longer periods of time (i.e., hours to days).…”

mentioning

confidence: 99%

Overcoming a Permeability Barrier by Microinjecting Cryoprotectants into Zebrafish Embryos (Brachydanio rerio)

2000

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Feasibility Studies on Vitrification of Intact Zebrafish (Brachydanio rerio) Embryos

Cited by 87 publications

References 0 publications

Cryopreservation of Primordial Germ Cells by Rapid Cooling of Whole Zebrafish (Danio rerio) Embryos

Cryopreservation of Primordial Germ Cells by Rapid Cooling of Whole Zebrafish (Danio rerio) Embryos

Toxicity of cryoprotectants on Prochilodus lineatus (Valenciennes, 1837) (curimba) embryos in an experimental incubator (Characiformes: Prochilodontidae)

Overcoming a Permeability Barrier by Microinjecting Cryoprotectants into Zebrafish Embryos (Brachydanio rerio)

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