Cryopreservation of testicular tissue from the dog (Canis familiaris) and wild boar (Sus scrofa) by slow freezing and vitrification: Differences in cryoresistance according to cell type

Picazo, C.M.; Castaño, C.; Bóveda, Paula; Toledano-Dı́az, A.; Velázquez, Rosario; Pequeño, Belén; Esteso, M.C.; Gadea, J.; Villaverde‐Morcillo, Silvia; Cerdeira, J.; Santiago-Moreno, J.

doi:10.1016/j.theriogenology.2022.07.020

Cited by 4 publications

(11 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Five tissue fragments from each testicle were threaded onto an acupuncture needle (0.18 x 30 mm; medical Device Co., Ltd. Wujiang, China) and plunged into DMEM F-12 with 1.06 M DMSO and 1.35 M ethylene glycol according to Picazo et al ( 17 ). After 10 min of equilibration, the needles were immersed in DMEM F-12 with 2.1 M DMSO, 2.7 M ethylene glycol, and 0.5 M sucrose for 2 min.…”

Section: Methodsmentioning

confidence: 99%

“…Slow freezing was performed following the protocol described by Picazo et al ( 17 ). Eight tissue fragments from each testicle were transferred to a 2 ml cryovial with 0.5 ml of DMEM F-12 supplemented with 20% FBS and 2.8 M dimethylsulfoxyde (DMSO).…”

Section: Methodsmentioning

confidence: 99%

“…Among wild species, comparative studies have been conducted in wild boar ( 17 ), red-rumped agouti ( 11 ), collared peccary ( 10 ), gray wolf ( 9 ), jungle cat, lion, leopard, rusa deer, fea's muntjac, and sumatran serow ( 21 ) but there are many wildlife species in which comparison of cryopreservation methods are still required to better understand the species-specific differences that lead to structural and functional germ cell damage. In wild boar and collared peccary both techniques are equally effective ( 10 , 17 ). Furthermore, two vitrification methods were also compared in the latter species, obtaining better results with conventional vitrification than with solid surface vitrification ( 10 ).…”

Section: Introductionmentioning

confidence: 99%

“…In boars and cats, slow freezing and vitrification showed similar results in terms of testicular cell survival ( 2 , 13 , 14 ). Conversely, in domestic rodents and dogs, vitrification was superior to slow freezing in preserving testicular cell integrity ( 2 , 15 – 17 ). In other studies, fast freezing led to greater testicular sperm viability than slow freezing or vitrification in cats ( 18 , 19 ), while slow freezing using dimethylsulfoxyde, ethylene glycol, and trehalose provided better results than vitrification in dogs ( 2 ).…”

Section: Introductionmentioning

confidence: 99%

“…In ram, slow freezing better preserved testicular morphology than vitrification ( 20 ). Among wild species, comparative studies have been conducted in wild boar ( 17 ), red-rumped agouti ( 11 ), collared peccary ( 10 ), gray wolf ( 9 ), jungle cat, lion, leopard, rusa deer, fea's muntjac, and sumatran serow ( 21 ) but there are many wildlife species in which comparison of cryopreservation methods are still required to better understand the species-specific differences that lead to structural and functional germ cell damage. In wild boar and collared peccary both techniques are equally effective ( 10 , 17 ).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

DNA integrity and viability of testicular cells from diverse wild species after slow freezing or vitrification

Peris-Frau

Benito-Blanco²,

Martínez‐Nevado³

et al. 2023

Front. Vet. Sci.

View full text Add to dashboard Cite

Introduction and objectiveCryopreservation of testicular tissues offers new possibilities to protect endangered species, genetically valuable individuals or even the fertility potential of prepubertal individuals who have died unexpectedly. However, the use of this technique still remains a challenge. In this study, slow freezing and vitrification of testicular tissue was investigated to find out which cryopreservation method could better preserve the viability and DNA integrity of testicular germ cells in diverse wild species.MethodsTestes were obtained post-mortem from 18 artiodactyls (wild boar, roe deer, dwarf goat, mhor gazelle, European mouflon, African forest buffalo, Malayan tapir, dorcas gazelle, Iberian ibex, gnu, red river hog), 5 primates (colobus monkey, capuchin monkey, mandrill), 8 carnivores (gray wolf, Persian leopard, binturong, European mink, American black bear, suricata), and 2 rodents (Patagonian mara). The testicles belonged to adult individuals and were cut into small pieces and cryopreserved by needle immersed vitrification or uncontrolled slow freezing using a passive cooling device. After warming or thawing, testicular tissues were enzymatically digested and two germ cell types were differentiated based on their morphology: rounded cells (spermatogonia, spermatocytes, and early spermatids) and elongated cells (elongated spermatids and spermatozoa). Cell viability was assessed by SYBR-14/propidium iodide while DNA fragmentation by TUNEL assay with fluorescence microscope.Results and discussionOur preliminary results revealed that our uncontrolled slow freezing method better preserved the viability and DNA integrity of elongated cells than vitrification. Such trend was observed in all species, being significant in artiodactyls, carnivores, and primates. Similarly, the viability and DNA integrity of rounded cells was also better maintained in primates by uncontrolled slow freezing, while in carnivores, vitrification by needle immersion showed better results in this type of cells. In artiodactyls and rodents both techniques preserved the viability of rounded cells in a similar manner, although the DNA integrity of these cells was greater after needle immersed vitrification in artiodactyls.ConclusionsIn conclusion, the effectiveness of each cryopreservation method is affected by the phylogenetic diversity between species and cell type.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%