A Preliminary Technique for the Isolation and Culture of Brown Trout (Salmo trutta macrostigma, Dumeril, 1858) Spermatogonial Stem Cell

Authors

  • Sehriban CEK-YALNIZ Faculty of Marine Science and Technology, Iskenderun Technical University
  • Kamuran Umut YARAS Faculty of Marine Science and Technology, Iskenderun Technical University

DOI:

https://doi.org/10.30564/jfsr.v1i2.1354

Abstract

This study was aimed to find a practical technique for isolation and culture spermatogonial stem cells from male brown trout (Salmo trutta macrostigma). Twelve wild juvenile male were obtained from Kılıç Trout Fish Farm (Kahramanmaraş, Turkey). The juveniles were taken alive to the aquaria unit and stored in a 1000-liter capacity fiberglass tank.  In order to identify the best size, age and testis structure of S.t. macrostigma for spermatogonial stem cell isolation and culture. Morphological and histological testis conditions were assessed. Fish were anesthetized with 0.04% 2-phenoxethanol. The surface of the fish was sterilized with 70% ethanol. Twelve fish were divided into two groups for enzyme digestion, and each group was divided into two replicates (three fish per replicate). Testis tissue of group one were digested by 0.25% trypsin- EDTA, and testis tissues of group two were digested by 0.05% trypsin-EDTA. At the end of the trial, first, the best age, size and weight of the male fish for spermatogonial stem cell isolation and culture were identified as 5+ month old, 12.13±1.5 cm, 19, 25±7.05 g respectively. Then, the highest spermatogonial stem cells were measured in the stage one and two of the testes. Finally, isolation and culture conditions were optimized for male S.t. macrostigma. Spermatogonial stem cell isolation and culture techniques were defined for fish in order to be used in surrogate reproduction technologies and gene transfer systems.

Keywords:

Histology; Age-Size; Trypsin-EDTA; Developmental stages

References

[1] Çek, Ş.. Early Gonadal Development and Sex Differentiation in Rosy Barb (Puntus Conhonius).Animal Biology, 2006, 56(3): 335-350.

[2] https://doi: 10.1163/157075606778441895

[3] Çek, Ş. and Yılmaz, E.. Gonad Development and Sex Ratio of Sharptooth Catfish (Clarias gariepinus Burchell, 1822) Cultured under Laboratory Conditions. Turkish Journal of Zoology, 2007, 31: 35-46.

[4] Okutsu, T., Suzuki, K., Takeuchi, Y., Takeuchi, T, and Yoshizaki, G.. Testicular germ cells can colonize sexually undifferentiated embryonic gonad and produce functional eggs in fish. Proceeding of the National Academy of Sciences of the United States of America, 2006, 103: 2725-2729.

[5] www.pnas.orgcgidoi10.1073pnas.0509218103

[6] Yoshizaki, G., Okutsu, T., Morita, T., Terasawa, M., Yazawa, R. and Takeuchi, Y.. Biological Characteristics of Fish Germ Cells and Their Applications to Developmental Biotechnology. Reproduction in Domestic Animals, 2012, 47(4): 187-92.

[7] https://doi:10.1111/j.1439-0531.2012. 02074

[8] Hong, N., Yuan, Y., Wang, T., Yi, W., Xu, H., Zeng, H., Song, J. and Hong, Y.. Dnd is a Critical Specifier of Primordial Germ Cells in The Medeka Fish. Stem Cell Reports. 2016, 6(3): 411-422.

[9] https://doi: 10.1016/j.stemcr.2016.01.002

[10] Shikina, S., Ihara, S. and Yoshizaki, G.. Culture conditions for maintaining the survival and mitotic activity of rainbow trout transplantable Type A spermatogonia. Molecular Reproduction Development, 2008, 75: 529-537.

[11] https://doi.org/10.1002/mrd.20771

[12] Yozhizaki,G., Fujinuma, K., Iwasaki, Y., Okutsu, T., Shikina, S., Yazawa, R. and Takeuchi, Y.. Spermatogonial Transplantation in Fish: A Novel Method for the Preservation of Genetic Resources. Comparative Biochemistry and Physiology, 2011, 6: 55–61.

[13] https://doi:10.1016/j.cbd.2010.05.003

[14] Turan, D., Kottelat, M. and Bektaş, Y.. Salmo tigridis, a New Species of Trout from the Tigris River, Turkey (Teleostei: Salmonidae). Zootaxa, 2011, 2993: 23-33.

[15] Turan, D., Doğan, E., Kaya, C. and Kanyılmaz, M.. Salmo Kottelati , a New Species of Trout from Alakır Stream, Draining to the Mediterranean in Southern Anatolia, Turkey (Teleostei, Salmonidae). ZooKeys, 2014, 462: 135-151.

[16] https://dx.doi.org/10.3897/zookeys.462.8177

[17] Tougard, C., Justy, F., Guinand, B., Douzery, E.J.P. and Berrebi, P.. Salmo trutta macrostigma (Teleostei, Salmonidae): Nothing More Than a Brown trout (S. Trutta) Lineage? Journal of Fish Biology. 2018, 93(2): 302-310.

[18] https:// doi: 10.1111/jfb.13751

[19] Kocabaş, M., Kayim, M., Can, E., Kutluyer, F. and Aksu, O.. The Reproduction Triats of Brown Trout (Salmo trutta macrostigma) Turkey Journal of Animal and Veterinary Advences. 2001, 10(13): 1632-1637.

[20] Tarkan, A.S., Gaygusuz, O., Ozulug, M. and Gaygusuz, C.G.. Reoccurrence of Salmo trutta macrostigma (Dumreil, 1858) in Lake Sapanca Basin (Sakarya, Turkey): Implications for Conservation. Journal of Fisheries and Aquatic Science, 2008, 3(1): 87-91.

[21] Alp, A. and Kara, C.. Ceyhan, Seyhan ve Fırat Havzalarındaki Doğal Alabalıklarda (Salmo trutta macrostigma Dumreil, 1858 ve Salmo Platycephalus Behnke, 1968) Boy, Ağırlık ve Kondüsyon Faktörleri E.U. Journal of Fisheries and Aquatic Science, 2004, 21: 9-15.

[22] Gülle, I., Küçük, F., Güçlü, S.S., Gümüş, E. and Demir, O.. Dağ Alabalığı (Salmo trutta macrostigma Dumreil, 1858)’nın Türkiye2nin Batı Akdeni,z Havzası’ındaki Yayılış Alanı, Populasyon ve Habitat Özellikleri , Türk Sucul Yaşam Dergisi, 2007, (5-8): 189-198.

[23] Çiçek, E., Fricke, R., Sungur, S. and Eagderi, S.. Endemic Freshwater Fishes of Turkey. Journal of Fish Taxonomy. 2018, 3(4): 1-39.

[24] Lujic, J., Marinovic, Z., Bajec, S.S., Djurdjevic, I., Urbanyi, B. and Horvath A.. Interspecific Germ Cell Transplantation: A new Light in the Conservation of Valuable Balkan Trout Genetic Resources. Fish Physiology and Biochemistry, 2018, 44(6): 1487-1498.

[25] https://doi.org/10.1007/s10695-018-0510-4

[26] Çek, Ş., Bromage, N., Randal, C. and Rana, K.. Oogenesis, Hemotosomatic and Gonadosomatic Indexes and Sex Ratio in Rosy Barb (Puntius Conchonius). Turkish Journal of Fisheries and Aquatic Science, 2001, 1: 33-41.

[27] Çek, Ş. and Yılmaz, E.. The Effect of Varying Dietary Energy on Gonad Development at First Sexual Maturity of the Sharp Catfish (Clarias gariepinus Burchell, 1822). Aquaculture International, 2009, 17: 553-563.

[28] https://doi.org/10.1007/s10499-008-9224-4

[29] Shang, M.. Isolation, identification, culture, cryopreservation, genetic transformation and transplantation of catfish germ line stem cells. PhD. Thesis, Auburn University, 2013: 163.

[30] Shang, M., Su, B., Lipke, E.A., Perera, D.A., Li. C., Qin, Z., Li, Y., Dunn. D.A., Çek, S., Peatman, E. and Dunham, R.A.. Spermatogonial Stem Cells Specific Marker Identification in Channel Catfish, Ictalurus punctatus and Blue Catfish, I. Furcatus. Fish Physiology and Biochemistry, 2015, 41(6): 1545-1556.

[31] https://doi.org/10.1007/s10695- 015-0106-1

[32] Shang, M., Su, B., Perera, D.A., Alsaqufi, A., Lipke, E.A., Çek, S., Dunn, D.A.Qin, Z., Peatman, E. and Dunham, R.A.. Testicular Germ Line Cell Identification, Isolation and Transplantation in Two North American Catfish Species. Fish Physiology and Biochemistry, 2018, 44(2): 717-733.

[33] https://doi.org/10.1007/s10695-018-0467-3

[34] Kise K, Yoshikawa H, Sato M, Tashiro M, Yazawa R, Nagasaka Y, Takeuchi Y. and Yoshizaki G.. Flow-cytometric isolation and enrichment of teleost type A spermatogonia based on light-scattering properties. Biology of Reproduction, 2012, 86: 1-12.

[35] https://doi.org/10.1095/biolreprod.111.093161

[36] Sato, M.,Hayashi, M. and Yoshizaki, G.. Stem cell activity of type A spermatogonia is seasonally regulated in rainbow trout. Biology of Reproduction, 2017, 96(6): 1303-1316.

[37] https://doi.org/10.1093/biolre/iox049

[38] Hayashi, M., Sato, M., Nagasaka, Y., Sadaie, S., Kobayashi, S. and Yoshizaki, G.. Enrichment of Spermatogonial Stem cells Using Side Population in Teleost. Biology of Reproduction, 2014, 91: 1-8.

[39] https://doi:10.1095/biolreprod.113.114140

[40] Lujic, J., Marinovic, Z., Bajec, S.S., Djurdjevic, I., Urbanyi, B. and Hotvath A.. First Successful Vitrification of Salmonoid Ovarian Tissue. Cryobiology, 2017, 76: 154-157.

[41] https://doi.org/10.1016/j.cryobiol.2017.04.005

[42] Marinović, Z., Lujić, J., Kása, E., Bernáth, G., Urbányi, B. and Horváth, Á.. Cryosurgical of isolated testicular cells and testicular tissue of tench Tinca tinca and goldfish Carassius auratus following slow-rate freezing. General and Comparative Endocrinology, 2017, 245: 77-83.

[43] https://doi.org/10.1016/j.ygcen.2016.07.005

[44] Lacerda, S.M.S.N., Martinez, E.R.M., Mura, I.L.D.D., Dorettoc, L.B., Costa, G.M.J.,Silva, M.A., Digmayer, M., Nobrega, R.H. and França, L.R.. Duration of spermatogenesis and identification of spermatogonial stem cell markers in a Neotropical catfish, Jundiá (Rhamdia quelen). General and Comparative Endocrinology, 2019, 274: 249-259.

[45] https://doi.org/10.1016/j.ygcen.2018.10.018

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Vol. 1 , Iss. 2 (October 2019)

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