Abstract

  Cell lines are useful tools to facilitate in vitro studies in virology, epizootology and many other biological field of research. We describe an efficient method of primary cell culture obtaining from pool of internal parenchymal organs of tadpoles Xenopus laevis. The enzymatically digested tissue attached well to previously treated cell culture surface and start of exponential cell growth ranged from 10 to 14 days. Cells had fibroblastic morphology and normal diploid karyotype up to ten passage. African clawed frog cell culture was propagated continuously, cryopreserved and recovered successfully. Acceptable cell growth occurred at 29°C in standard medium DMEM after osmotic pressure correction and FBS adding. This cell culture would be used in the study of viral diseases of amphibian, as well as fundamental virological and epizootological investigation.

Keywords: African Clawed Frog (Xenopus laevis); Cell Culture; Amphibian; Fibroblast

References

1. Alford R and Richards SJ. “Global amphibian declines: A problem in applied ecology”. Annual Review of Ecology, Evolution, and Systematics 30 (1999): 133-165.
2. Flores Nava A. FAO Inland Water Resources and Aquaculture Service (FIRI) Cultured Aquatic Species Information Programme - Rana catesbeiana. Cultured Aquatic Species Fact Sheets”. (2005).
3. Schlaepfer M., et al. “Challenges in evaluating the impact of the trade in amphibians and reptiles on wild populations”. BioScience3 (2005): 256-264.
4. FAO Fishery Information, Data and Statistics Unit (FAO-FIDI). Collation, analysis and dissemination of global and regional fishery statistics. FI Programme Websites. FAO – Rome (2004).
5. Kusrini M., et al. “Indonesia’s exports of frogs’ legs”. Traffic Bulletin1 (2006): 13-24.
6. Niekisch M. “The international trade of frogs’ legs”. Traffic Bulletin 8 (1986): 7-10.
7. Lau M., et al. “Wildlife Trade in Southern China including Hong Kong and Macao”. Report to Biodiversity Working Group of China Council for International Cooperation on Environment and Development Projects (1995).
8. Rueda-Almonacid J. “Status and threats produced by the introduction of the bullfrog in Columbia”. Revista de la academia Colombiana de ciencias exactas fisicas y naturales 23 (2000): 367-393.
9. Kiesecker J., et al. “Potential mechanisms underlying the displacement of native red-legged frogs by introduced bullfrogs”. Ecology 82 (2001): 1964-1970.
10. Mazzoni R., et al. “Emerging pathogen of wild amphibians in frogs (Rana catesbeiana) farmed for international trade”. Emerging Infectious Diseases8 (2003): 995-998.
11. Goris R., et al. “Guide to the amphibians and reptiles of Japan”. Kreiger Publishing Company, Malabar, Florida, USA (2004).
12. Gasc, J., et al. “Rana catesbeiana Shaw, 1802” Atlas of Amphibians and Reptiles in Europe (1997): 132-133.
13. The OIE Worldwide Monitoring System for Wild Animal Diseases [Electronic resource]. The World Organisation for Animal Health (OIE).
14. Dobson A., et al. “Biosecurity: The Socio-Politics of Invasive Species and Infectious Diseases”. Routledge, London (2013).
15. Freshney R Ian “Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications” (2016).
16. Philippe G., et al. “Recent developments in the field of arrow and dart poisons”. Journal of Ethnopharmacology 100 (2005): 85-91.
17. Clarke B. “The natural history of amphibian skin secretions, their normal functioning and potential medical applications”. Biological Reviews of the Cambridge Philosophical Society 3 (1997): 365-379.
18. Pukala T., et al. “Host-defence peptides from the glandular secretions of amphibians: structure and activity”. Natural Product Reports 23 (2006): 368-393.
19. VanCompernolle S., et al. “Antimicrobial peptides from amphibian skin potently inhibit Human Immunodeficiency Virus infection and transfer of virus from dendritic cells to T cells”. Journal of Virology18 (2005): 11598-11606.
20. George W Nace. “Amphibians: Guidelines for the Breeding, Care and Management of Laboratory Animals”. National Academy of Sciences Washington, D. C (1974).
21. Rothblat G., et al. “Growth, Nutrition and Metabolism of Cells in Culture”. Acad Press, INC. London LTD (1972).
22. Davis J. “Basic cell culture. Practical approach”. 2nd ed. Oxford: Univ. Press (2001).
23. Nie W., et al. “The genome phylogeny of domestic cat, red panda and five mustelid species revealed by comparative chromosome painting and G-banding”. Chromosome Research 10 (2002): 209-222.
24. Anizet M., et al. “Characterization of a new cell line, XL2, obtained from Xenopus laevis and determination of optimal culture conditions”. In Vitro 17 (1981): 267-274.
25. Asashima M., et al. “Purification of mesodermal inducing substance and protein synthesis using this material”. Progress in Clinical and Biological Research 226 (1986): 55-66.
26. Fukui A., et al. “A new cell line (XTY) from a tumor of Xenopus laevis”. Experientia 48 (1992): 87-91.
27. Smith J., et al. “Xenopus cell lines”. Methods Cell Biology 36 (1991): 635-654.
28. Balls M., et al. “Amphibian cells in vitro II. Effects of variations in medium osmolarity on a permanent cells line isolated from Xenopus”. Experimental Cell Research 76 (1973): 333-336.
29. Ruoslahti E., et al. “New perspectives in cell adhesion: RGD and integrins”. Science4826 (1987): 491-497.
30. van der Merwe M., et al. “Investigating the establishment of primary cell culture from different abalone (Haliotis midae) tissues”. Cytotechnology 62 (2010): 265-277.
31. Ellinger M., et al. “Amphibian cell culture: established fibroblastic line from embryos of the discoglossid frog, Bombina orientalis”. In Vitro 19 (1983): 429-434.

Citation

Citation: Zinaida Klestova and Iryna Savinova. “Efficient Establishment of Cell Culture from Xenopus laevis as a Model for In Vitro Studies".Acta Scientific Microbiology 3.1 (2020): 42-46.

Copyright

Copyright: © 2020 Lia Zinaida Klestova and Iryna Savinova. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.