Preservation of common carp germ cells under hypothermic conditions: Whole tissue vs isolated cells

Lujić, Jelena; Marinović, Zoran; Kása, Eszter; Šćekić, Ilija; Urbányi, Béla; Horváth, Ákos

doi:10.1111/rda.13220

Cited by 3 publications

(2 citation statements)

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“…Additionally, cryopreservation gives a possibility to synchronize and carry out transplantation according to the availability of hosts. The results of the present study can be used in combination with the hypothermic storage described by Lujić et al [59] where hypothermic storage is optimal for short-term storage of up to two weeks, while the freezing methodology developed in this study is optimal for long-term storage. Further steps will be taken to develop a protocol for female germ cell cryopreservation as well to improve transplantation success using younger recipients or developing germ-less carp hosts for allogenic germ cell transplantation.…”

Section: Resultsmentioning

confidence: 85%

Cryopreservation and transplantation of common carp spermatogonia

et al. 2019

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Common carp ( Cyprinus carpio ) is one of the most cultured fish species over the world with many different breeds and plenty of published protocols for sperm cryopreservation. However, data regarding preservation of gonadal tissue and surrogate production is still missing. A protocol for freezing common carp spermatogonia was developed through varying different factors along a set of serial subsequent experiments. Among the six cryoprotectants tested, the best survival was achieved with dimethyl sulfoxide (Me 2 SO). In the next experiment, a wide range of cooling rates (0.5–10°C/min) and different concentrations of Me 2 SO were tested resulting in the highest survival achieved using 2 M Me 2 SO and cooling rate of -1°C/min. When testing different tissue sizes and incubation times in the cryomedia, the highest viability was observed when incubating 100 mg tissue fragments for 30 min. Finally, sugar supplementation did not yield significant differences. When testing different equilibration (ES) and vitrification solutions (VS) used for needle-immersed vitrification, no significant differences were observed between the tested groups. Additionally, varied exposure time to VS did not improve the vitrification outcome where the viability was 4-fold lower than that of freezing. The functionality of cryopreserved cells was tested by interspecific transplantation into sterilized goldfish recipients. The exogenous origin of the germ cells in gonads of goldfish recipient was confirmed by molecular markers and incorporation rate was over 40% at 3 months post-transplantation. Results of this study can serve for long-term preservation of germplasm in carp which can be recovered in a surrogate recipient.

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Section: Resultsmentioning

confidence: 85%

Cryopreservation and transplantation of common carp spermatogonia

et al. 2019

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“…hosts. The results of the present study can be used in combination with the hypothermic storage 471 described by Lujić et al (2018) where hypothermic storage is optimal for short-term storage of 472 up to two weeks, while the freezing methodology developed in this study is optimal for long-473 term storage [58]. Further steps will be taken to develop a protocol for female germ cell 474 cryopreservation as well to improve transplantation success using younger recipients or 475 developing germ-less carp hosts for allogenic germ cell transplantation.…”

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confidence: 84%

Cryopreservation and transplantation of common carp spermatogonia

Franěk

Marinović

Lujić

et al. 2018

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20Common carp (Cyprinus carpio) is one of the most cultured fish species over the world with 21 many different breeds and plenty of published protocols for sperm cryopreservation, however, 22 data regarding preservation of gonadal tissue and surrogate production is still missing. 23 A protocol for freezing common carp spermatogonia was developed through varying different 24 factors along a set of serial subsequent experiments. Among the six cryoprotectants tested, the 25 best survival was achieved with dimethyl sulfoxide (Me 2 SO). In the next experiment, a wide 26 range of cooling rates (0.5 -10 ˚C/min) and different concentrations of Me 2 SO were tested 27 resulting in the highest survival using 2 M Me 2 SO and cooling rate of -1 ˚C/min. When testing 28 different tissue sizes and incubation times in the cryomedium, the highest viability was 29 observed when incubating 100 mg tissue fragments for 30 min. Finally, sugar supplementation 30 did not yield significant differences. When testing different equilibration (ES) and vitrification 31 solutions (VS) used for needle-immersed vitrification, no significant differences were observed 32 between the tested groups. Additionally, varied exposure time to VS did not improve the 33 vitrification outcome where the viability was 4-fold lower than that of freezing. The 34 functionality of cryopreserved cells was tested by interspecific transplantation into sterilized 35 goldfish recipients. The exogenous origin of the gonads in goldfish recipients was confirmed 36 by molecular markers and incorporation rate was over 40% in both groups at 3 months post 37 transplantation. Results of this study can serve as an alternative way for long-term preservation 38 of germplasm in carp which can be recovered in a surrogate recipient. 39 Introduction 42Common carp (Cyprinus carpio) is one of the oldest domesticated fish species in the world and 43 is mainly cultured in Europe and Asia. Nowadays, the common carp expanded to all continents 44 with exception of Antarctica. Overall carp production from aquaculture in 2014 was more than 45 4 million tons -around 14% of the total freshwater aquaculture production in that year [1]. 46 Fruitful history and lasting popularity of this species gave to rise many different strains and 47 lines which became important for breed management and production of hybrids in Central 48 Europe [2-4]. Due to this fact, significant efforts have been committed to preservation of carp 49 genetic resources. Long-term cultivation of pure-breed livestock [5], methods for genetic 50 diversity identification [6-9] as well as methods for creating and keeping gene banks through 51 sperm cryopreservation [10-14] have been developed. However, ex situ preservation of 52 valuable genetic material still relies only on sperm cryopreservation. 53 Recent progress in biotechnology revealed a very efficient alternative for preservation and 54 restoration of a valuable genetic material using cryopreservation of germline stem cells and 55 surrogate offspring production. Ge...

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