Development of a spermatogonia cryopreservation protocol for blue catfish, Ictalurus furcatus

Muyassar Abualreesh; Jaelen N Myers; Jeremy Gurbatow; Andrew Johnson; De Xing; Jinhai Wang; Shangjia Li; Michael Coogan; Khoi Vo; Nour El Husseini; Rex A Dunham; Ian A E Butts

doi:10.1016/j.cryobiol.2020.10.010

Development of a spermatogonia cryopreservation protocol for blue catfish, Ictalurus furcatus

Cryobiology. 2020 Dec:97:46-52. doi: 10.1016/j.cryobiol.2020.10.010. Epub 2020 Oct 13.

Authors

Affiliations

¹ School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL, 36849, USA.
² School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL, 36849, USA; Department of Aquaculture and Technology, Can Tho Technical Economic College, Can Tho, Viet Nam.
³ School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL, 36849, USA; Department of Animal and Avian Sciences, University of Maryland, College Park, MD, 20742, USA.
⁴ School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL, 36849, USA. Electronic address: iab0007@auburn.edu.

PMID: 33058900
DOI: 10.1016/j.cryobiol.2020.10.010

Abstract

Sustainability of channel catfish, Ictalurus punctatus ♀ × blue catfish, Ictalurus furcatus ♂ hybrid aquaculture relies on new innovative technologies to maximize fry output. Transplanting spermatogonial stem cells (SSCs) from blue catfish into channel catfish hosts has the potential to greatly increase gamete availability and improve hybrid catfish fry outputs. Cryopreservation would make these cells readily accessible for xenogenesis, but a freezing protocol for blue catfish testicular tissues has not yet been fully developed. Therefore, the objectives of this experiment were to identify the best permeating [dimethyl sulfoxide (DMSO), ethylene glycol (EG), glycerol, methanol] and non-permeating (lactose or trehalose with egg yolk or BSA) cryoprotectants, their optimal concentrations, and the best freezing rates (-0.5, -1.0, -5.0, -10 °C/min until -80 °C) that yield the highest number of viable type A spermatogonia cells. Results showed that all of these factors had significant impacts on post-thaw cell production and viability. DMSO was the most efficient permeating cryoprotectant at a concentration of 1.0 M. The optimal concentration of each cryoprotectant depended on the specific cryoprotectant due to interactions between the two factors. Of the non-permeating cryoprotectants, 0.2 M lactose with egg yolk consistently improved type A spermatogonia production and viability beyond that of the 1.0 M DMSO control. The overall best freezing rate was consistent at -1 °C/min, but similar results were obtained using -0.5 °C/min. Overall, we recommend cryopreserving blue catfish testicular tissues in 1.0 M DMSO with 0.2 M lactose and egg yolk at a rate of either -0.5 or -1 °C/min to achieve the best cryopreservation outcomes. Continued development of cryopreservation protocols for blue catfish and other species will make spermatogonia available for xenogenic applications and genetic improvement programs.

Keywords: Cryopreservation; Germplasm; Selective breeding; Type A spermatogonia; Xenogenesis.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Catfishes*
Cryopreservation / methods
Cryoprotective Agents / pharmacology
Dimethyl Sulfoxide
Ictaluridae*
Male
Semen Preservation* / veterinary
Spermatogonia
Spermatozoa

Substances

Cryoprotective Agents
Dimethyl Sulfoxide