Preservation of female genetic resources of common carp through oogonial stem cell manipulation

Roman Franěk, Tomáš Tichopád, Christoph Steinbach, Xuan Xie, Jelena Lujić, Zoran Marinović, A. Horváth, Vojtěch Kašpar, Martin Pšenička

Research output: Article

1 Citation (Scopus)

Abstract

Several experiments were conducted in order to develop an optimal protocol for slow-rate freezing (−1 °C/min) and short-term storage (−80 or 4 °C) of common carp ovarian tissue fragments with an emphasis on oogonial stem cells (OSCs). Dimethyl sulfoxide (Me2SO) with concentration of 1.5 M was identified as the best cryoprotectant in comparison to propylene glycol and methanol. When comparing supplementation of sugars (glucose, trehalose, sucrose) in different concentrations (0.1, 0.3, 0.5 M), glucose and trehalose in 0.3 M were identified as optimal. Short-term storage options for ovarian tissue pieces at −80 °C and 4 °C were tested as alternatives to cryopreservation and storage in liquid nitrogen. The presence of OSCs was confirmed by immunocytochemistry and viability after storage was determined by the trypan blue exclusion test. This study identified the optimal protocol for OSC cryopreservation using slow rate freezing resulting in ∼65% viability. The frozen/thawed OSCs were labelled by PKH-26 and transplanted into goldfish recipients. The success of the transplantation was confirmed by presence of fluorescent cells in the recipient gonad and later on by RT-PCR with carp dnd1 specific primers. The results of this study can facilitate long-term preservation of common carp germplasm which can be recovered in a surrogate recipient through interspecific germ cell transplantation.

Original languageEnglish
JournalCryobiology
DOIs
Publication statusPublished - jan. 1 2019

Fingerprint

Carps
Stem cells
Cyprinus carpio
genetic resources
Trehalose
Cryopreservation
trehalose
Freezing
cryopreservation
freezing
viability
Tissue
Glucose
Propylene Glycol
cell transplantation
glucose
Goldfish
propylene glycol
Trypan Blue
goldfish

Keywords

    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)
    • Agricultural and Biological Sciences(all)

    Cite this

    Franěk, R., Tichopád, T., Steinbach, C., Xie, X., Lujić, J., Marinović, Z., ... Pšenička, M. (2019). Preservation of female genetic resources of common carp through oogonial stem cell manipulation. Cryobiology. https://doi.org/10.1016/j.cryobiol.2019.01.016

    Preservation of female genetic resources of common carp through oogonial stem cell manipulation. / Franěk, Roman; Tichopád, Tomáš; Steinbach, Christoph; Xie, Xuan; Lujić, Jelena; Marinović, Zoran; Horváth, A.; Kašpar, Vojtěch; Pšenička, Martin.

    In: Cryobiology, 01.01.2019.

    Research output: Article

    Franěk, R, Tichopád, T, Steinbach, C, Xie, X, Lujić, J, Marinović, Z, Horváth, A, Kašpar, V & Pšenička, M 2019, 'Preservation of female genetic resources of common carp through oogonial stem cell manipulation' Cryobiology. https://doi.org/10.1016/j.cryobiol.2019.01.016
    Franěk, Roman ; Tichopád, Tomáš ; Steinbach, Christoph ; Xie, Xuan ; Lujić, Jelena ; Marinović, Zoran ; Horváth, A. ; Kašpar, Vojtěch ; Pšenička, Martin. / Preservation of female genetic resources of common carp through oogonial stem cell manipulation. In: Cryobiology. 2019.
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    AU - Marinović, Zoran

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