VIABILITY OF FIBROBLASTS OF TUGAI DEER (CERVUS HANGLU BACTRIANUS LYDEKKER, 1900) DURING CRYOPRESERVATION BY DIFFERENT METHODS

Main Article Content

Authors

T.T. Nurkenov

Institute of Zoology, Al-Farabi Ave., 93, 050060, Almaty, Kazakhstan

Y.A. Zima

Institute of Zoology, Al-Farabi Ave., 93, 050060, Almaty, Kazakhstan

B.S. Katubayeva

Institute of Zoology, Al-Farabi Ave., 93, 050060, Almaty, Kazakhstan

A.A. Grachev

Institute of Zoology, Al-Farabi Ave., 93, 050060, Almaty, Kazakhstan

Y.R. Baidavletov

Institute of Zoology, Al-Farabi Ave., 93, 050060, Almaty, Kazakhstan

D.Y. Toishybek

Institute of Zoology, Al-Farabi Ave., 93, 050060, Almaty, Kazakhstan

M.Y. Salmenova

Institute of Zoology, Al-Farabi Ave., 93, 050060, Almaty, Kazakhstan

Y.A. Assanova

LLP EMBRYO TECHNOLOGY LABS, Bogenbay batyr st., 149, 050012, Almaty, Kazakhstan

Y.M. Toishibekov

LLP EMBRYO TECHNOLOGY LABS, Bogenbay batyr st., 149, 050012, Almaty, Kazakhstan

Abstract

Currently, there is a rapid and continuous decline in wild animal populations, which threatens accelerated loss of biodiversity on a global scale. Along with in situ conservation strategies, ex situ conservation measures such as germplasm cryopreservation should be widely applied. Cryopreservation in combination with assisted reproductive technologies can become an important means of successfully preserving the biodiversity of the animal world. Methods based on obtaining cell cultures are of great importance in scientific research. The resulting cell lines can be frozen and stored in containers with liquid nitrogen for many years. Such cell and tissue biobanks represent a viable source of genetic material and living cells that offer numerous opportunities for further research.

The aim of this study was to investigate the scientific basis for cryopreservation of somatic cells of tugai deer, which is a species in need of special protection. Studies were conducted on the viability of tugai deer fibroblasts using various cryopreservation methods. The viability of frozen-thawed tugai deer fibroblasts was determined, and it was shown that the most effective cryopreservation of this species fibroblasts was the use of a cryoprotectant 1.5 M ethylene glycol and a slow equilibrium freezing mode using a programmed freezer.

Keywords

tugai deer, fibroblasts, slow freezing, vitrification, biodiversity, germplasm.

Article Details

References

Alexandre Rodrigues Silva, Gabriela Liberalino Lima, Gislayne Christianne Xavier Peixoto & Ana Liza Paz Souza (2015) Cryo-preservation in mammalian conservation biology: current applications and potential utility, Research and Reports in Bio-diversity Studies, 1-8, DOI: 10.2147/RRBS.S54294.

Y. M. Toishibekov, A. S. Kazybayeva, Y. A. Assanova, T. T. Nurkenov and D. Y. Toishybek. The effect of various cryoprotective agents and slow cooling rate on viability of goat ovarian tissue. Reproduction, Fertility and Development, 2021, 34(2), 257-257, Crossref.

Joppa LN, O’Connor B, Visconti P, Smith C, Geldmann J, Hoffmann M, et al. Big Data and biodiversity. Filling in biodiversity threat gaps. Science 2016;352(6284):416e8. Crossref.

Snyder NFR. Limitations of captive breeding in endangered species recovery. 1996.

Guidelines on the use of ex situ management for species conservation. Version 2.0. 2014.

Hildebrandt TB, Hermes R, Goeritz F, Appeltant R, Colleoni S, de Mori B, Diecke S, Drukker M, Galli C, Hayashi K, Lazzari G, Loi P, Payne J, Renfree M, Seet S, Stejskal J, Swegen A, Williams SA, Zainuddin ZZ, Holtze S. The ART of bringing extinction to a freeze - History and future of species conservation, exemplified by rhinos. Theriogenology. 2021 Jul 15;169:76-88. doi: 10.1016/j.theriogenology.2021.04.006. Epub 2021 Apr 18. PMID: 33940218.

Callaway E. Stem-cell plan aims to bring rhino back from brink of extinction. Nature 2016;533(7601):20e1.Crossref.

Hikabe O, Hamazaki N, Nagamatsu G, Obata Y, Hirao Y, Hamada N, et al. Reconstitution in vitro of the entire cycle of the mouse female germ line. Nature 2016;539(7628):299e303. Crossref.

Ryder OA, Onuma M. Viable cell culture banking for biodiversity characterization and conservation. Annu. Rev. Anim. Biosci. 2018;6:83e98. Crossref.

Comizzoli P, Songsasen N, Hagedorn M, Wildt DE. Comparative cryobiological traits and requirements for gametes and gonadal tissues collected from wildlife species. Theriogenology 2012;78(8):1666e81.Crossref.

Fernandes Pereira, Alexsandra, Lhara Ricarliany Medeiros de Oliveira, Leonardo Vitorino Costa de Aquino, João Vitor da Silva Viana, and Luanna Lorenna Vieira Rodrigues. 2023. ‘Strategies for the Establishment of Fibroblastic Lines for the Conserva-tion of Wild Mammals’. Veterinary Medicine and Science. IntechOpen. doi:10.5772/intechopen.114028.

Kuzˇmova´ E, Bartosˇ L, Kotrba R, Bubenik GA (2011) Effect of Different Factors on Proliferation of Antler Cells, Cultured In Vitro. PLoS ONE 6(3): e18053. doi:10.1371/journal.pone.0018053.

Magalhães LC, Bhat MH, Freitas JLS, Melo LM, Teixeira DIA, Pinto LCA, Câmara LMC, Duarte JMB, Freitas VJF. The Effects of Cryopreservation on Different Passages of Fibroblast Cell Culture in Brown Brocket Deer (Mazama gouazoubira). Biopreserv Biobank. 2017 Oct;15(5):463-468. doi: 10.1089/bio.2017.0060. Epub 2017 Sep 18. PMID: 28922611.

Fernández-Santos MR, Esteso MC, Montoro V, Soler AJ, Garde JJ. Influence of various permeating cryoprotectants on freeza-bility of Iberian red deer (Cervus elaphus hispanicus) epididymal spermatozoa: effects of concentration and temperature of addition. J Androl. 2006 Nov-Dec;27(6):734-45. doi: 10.2164/jandrol.106.000505. Epub 2006 Jun 14. PMID: 16775252.

Leibo SP, Songsasen N. Cryopreservation of gametes and embryos of nondomestic species. Theriogenology 2002;57(1):303e26. Crossref.

Comizzoli P, Holt WV. Recent advances and prospects in germplasm preservation of rare and endangered species. Adv Exp Med Biol 2014;753:331e56. Crossref.

Brook, S.M., Donnithorne-Tait, D., Lorenzini, R., Lovari, S., Masseti, M., Pereladova, O., Ahmad, K. & Thakur, M. 2017. Cervus hanglu (amended version of 2017 assessment). The IUCN Red List of Threatened Species 2017: e.T4261A120733024. Crossref.

Praxedes EA, Borges AA, Santos MVO, Pereira AF. Use of somatic cell banks in the conservation of wild felids. Zoo Biology. 2018;37:258-263. DOI: 10.1002/zoo.21416.

Lhara R.M. De Oliveira, Érika A. Praxedes, Maria B. Silva, Leandro R. Ribeiro, Herlon V.R. Silva & Alexsandra F. Pereira. Comparative effect of cryoprotectant combinations on the conservation of somatic cells derived from jaguar, Panthera onca, towards the formation of biologic banks. An Acad Bras Cienc (2021) 93(4): e20190314. DOI 10.1590/0001-3765202120190314.

Li Y, Lu RH, Luo GF, Pang WJ, Yang GS. Effects of different cryoprotectants on the viability and biological characteristics of porcine preadipocyte. Cryobiology. 2006 Oct;53(2):240-7. doi: 10.1016/j.cryobiol.2006.06.007. PMID: 16930580.

Moulavi F, Hosseini SM, Tanhaie-Vash N, Ostadhosseini S, Hosseini SH, Hajinasrollah M, Asghari MH, Gourabi H, Shahverdi A, Vosough AD, Nasr-Esfahani MH. Interspecies somatic cell nuclear transfer in Asiatic cheetah using nuclei derived from post-mortem frozen tissue in absence of cryo-protectant and in vitro matured domestic cat oocytes. Theriogenology. 2017 Mar 1;90:197-203. doi: 10.1016/j.theriogenology.2016.11.023. Epub 2016 Nov 27. PMID: 28166968.

León-Quinto T, Simón MA, Cadenas R, Martínez A, Serna A. Different cryopreservation requirements in foetal versus adult skin cells from an endangered mammal, the Iberian lynx (Lynx pardinus). Cryobiology. 2014 Apr;68(2):227-33. doi: 10.1016/j.cryobiol.2014.02.001. Epub 2014 Feb 12. PMID: 24530371.

Benkeddache, D., Bodinier, P., Joly, T. et al. Recovery of viable cells from rabbit skin biopsies after storage at −20°C for up to 10 days. Cell Tissue Bank 13, 479–486 (2012). Crossref.

Zieger MA, Tredget EE, McGann LE. Mechanisms of cryoinjury and cryoprotection in split-thickness skin. Cryobiology. 1996 Jun;33(3):376-89. doi: 10.1006/cryo.1996.0038. PMID: 8689894.

Gandolfi F, Paffoni A, Papasso Brambilla E, Bonetti S, Brevini TA, Ragni G. Efficiency of equilibrium cooling and vitrification procedures for the cryopreservation of ovarian tissue: comparative analysis between human and animal models. Fertil Steril. 2006 Apr;85 Suppl 1:1150-6. doi: 10.1016/j.fertnstert.2005.08.062. PMID: 16616087.

Agca Y, Mullen S, Liu J, Johnson-Ward J, Gould K, Chan A, Critser J. Osmotic tolerance and membrane permeability charac-teristics of rhesus monkey (Macaca mulatta) spermatozoa. Cryobiology. 2005 Aug;51(1):1-14. doi: 10.1016/j.cryobiol.2005.04.004. PMID: 15922321.