New paper accepted in Cyobiology

A new paper in collaboration with our colleagues from University of South Bohemia in Česke Budejovice, Faculty of Fisheries and Protection of Waters (Czech Republic) has been accepted for publication in Cryobiology.

 

Roman Franěk, Tomáš Tichopád, Christoph Steinbach, Xuan Xie, Jelena Lujić, Zoran Marinović, Ákos Horváth, Vojtěch Kašpar, Martin Pšenička. Preservation of female genetic resources of common carp through oogonial stem cell manipulation. Cryobiology. DOI:10.1016/j.cryobiol.2019.01.016

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 oogonial stem cells (OSCs). Dimethyl sulfoxide 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, 0.5M), glucose and trehalose in 0.3 M were identified as optimal. Finally, different equilibration times revealed a positive impact of prolonged equilibration on post-thaw viability. 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 oogonia was confirmed by immunocytochemistry and viability after storage was determined by the trypan blue exclusion test. This study identified the best 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 vasa specific primers.