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Cell cultures derived from embryos and melanoma of poeciliid fish

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Summary

In poeciliid fish, melanoma of different degrees of malignancy can be produced by crossing specific genotypes. For a detailed investigation of the processes leading to proliferation or differentiation of the melanoma cells, it is necessary to establish cell cultures. The aim of the present study was to find out the optimal conditions for initiating and culturing poeciliid fish cells for the purpose of establishing cell cultures of melanoma. The optimal method was developed by using small pieces of late embryos as starting material and includes: (a) dispersion of tissue by mild stepwise treatment with a trypsin-EDTA mixture at low temperature; (b) culture of cells in the complex medium 199; (c) supplementation of medium with high percentage (20%) of fetal bovine serum; and (d) stabilization of pH by buffering the medium with HEPES. Under these conditions, primary and secondary cultures of embryonic cells have been initiated. An epithelial-like cell line has been subcultured for more than 80 passages. The method developed for embryonic tissues was used to start cell cultures from melanoma of platyfish-swordtail hybrids. Until now, only cells of rapidly growing malignant albino melanoma could be maintained in primary cultures. Secondary cultures could not be initiated since the melanoma cells tended to differentiate and stopped growing before a confluent monolayer was formed.

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References

  1. Kosswig, C. 1928. Über Kreuzungen zwischen den Teleostiern Xiphophorus helleri und Platypoecilus maculatus. I. Z. Indukt. Abstamm.-Vererbungsl. 47: 150–158.

    Article  Google Scholar 

  2. Häussler, G. 1928. Über Melanombildungen bei Bastarden von Xiphophorus helleri and Platypoecilus maculatus var. rubra. Klin. Wochenschr. 7: 1561.

    Article  Google Scholar 

  3. Gordon, M. 1931. The herediary basis of melanosis in hybrids of Mexican killifishes. Proc. Nat. Acad. Sci. U.S.A. 17: 276–280.

    Article  CAS  Google Scholar 

  4. Gordon, M. 1959. The melanoma cell as an incompletely differentiated pigment cell. In: M. Gordon (Ed.),Pigment Cell Biology. Academic Press, New York, pp. 215–239.

    Google Scholar 

  5. Anders, F. 1967. Tumor formation in platyfishswordtail hybrids as a problem of gene regulation. Experientia 23: 1–10.

    Article  PubMed  CAS  Google Scholar 

  6. Anders, A., F. Anders, and K. Klinke. 1973. Regulation of gene expression in the Gordon-Kosswig melanoma system. I. The distribution of the controlling genes in the genome of the Xiphophorin fish, Platypoecilus maculatus and Platypoecilus variatus. In: J. H. Schröder (Ed.),Genetics and Mutagenesis of Fish. Springer, Berlin-Heidelberg-New York, pp. 33–52.

    Google Scholar 

  7. Vielkind, J., U. Vielkind, and F. Anders. 1971. Melanotic and amelanotic melanomas in xiphophorin fish. Cancer Res. 31: 868–875.

    Google Scholar 

  8. Vielkind, U. 1976. Genetic control of cell differentiation in platyfish-swordtail melanomas. J. Exp. Zool. 196: 197–203.

    Article  PubMed  CAS  Google Scholar 

  9. Vielkind, U., W. Schlage, and F. Anders. 1977. Melanogenesis in genetically determined pigment cell tumros of platyfish and platyfish-swordtail hybrids: Correlation between tyrosinase activity and degree of malignancy. Z. Krebsforsch. 90: 285–299.

    Article  CAS  Google Scholar 

  10. Clark, H. F. 1972. Cultivation of cells from poikilothermic vertebrates. In: G. H. Rothblat and V. J. Cristofalo (Eds.),Growth. Nutrition and Metabolism of Cells in Culture. Vol. 2, Academic Press, New York, pp. 287–325.

    Google Scholar 

  11. Wolf, K., and M. C. Quimby. 1969. Fish cell and tissue culture. In: W. S. Hoar, and D. J. Randall (Eds.),Fish Physiology. Vol. 3, Academic Press, New York, pp. 253–305.

    Google Scholar 

  12. Grützner, L. 1958. In vitro-Züchtung des Leber-und Nierengewebes von Tinca vulgaris Cuv. (Schleie) in trypsinierten Einschichtgewebekulturen. Zentralbl. Bakteriol. Parasitenkd., Infektionskr. Hyg. Abt. I. Org. 173: 195–202.

    Google Scholar 

  13. McFalls, F. D., J. E. Markee, G. D. Wilbanks. 1967. The culture of adult and embryonic fish tissue. In Vitro 3: 196.

    Google Scholar 

  14. Kelly, R. K., and P. C. Loh. 1973. Some properties of an established fish cell line from Xiphophorus helleri (red swordtail). In Vitro 9: 73–79.

    Article  PubMed  CAS  Google Scholar 

  15. Grand, C. G., M. Gordon, and C. Cameron. 1941. Neoplasm studies. VIII. Cell types in tissue culture of fish melanotic tumors compared with mammalian melanomas. Cancer Res. 1: 660–666.

    CAS  Google Scholar 

  16. Grand, C. G., and G. Cameron. 1948. Tissue culture studies of pigmented melanomas: Fish, mouse, and human. In:The Biology of Melanomas. Spec. Publ. N. Y. Acad. Sci. 4: 171–176.

  17. Greenberg, S., M. J. Kopac, and M. Gordon. 1956. Cytology and cytochemistry of melanoma cells. Ann. N. Y. Acad. Sci. 67: 55–122.

    Article  CAS  Google Scholar 

  18. Tavolga, W. N. 1949. Embryonic development of the platyfish (Platypoecilus), the swordtail (Xiphophorus) and their hybrids. Bull. Am. Mus. Nat. Hist. 94: 161–230.

    Google Scholar 

  19. Günther, A. 1866.Catalogue of the Physostomi, Containing the Families Salmoniedae, Percopsidae, Galaxidae, Mormyridae, Gymnarchidae, Esocidae, Umbridae, Scombresocidae, Cyprinodontidae. In: Collection of the British Museum, London.

  20. Jordan, D. S., and B. W. Evermann. 1896. The fishes of North and Middle America. Part I. Bull. U. S. Nat. Mus. 47: 630–702.

    Google Scholar 

  21. Hausman, R. E., and A. A. Moscona. 1976. Isolation of retina-specific cell-aggregating factor from membranes of embryonic neural retina tissue. Proc. Natl. Acad. Sci. U.S.A. 73: 3594–3598.

    Article  PubMed  CAS  Google Scholar 

  22. Hayashi, M., Y. Ojima, and N. Asano. 1976. A cell line from teleost fish: Establishment and cytogenetical characterization of the cells. Jpn. J. Genet. 51: 65–68.

    Google Scholar 

  23. Morgan, J. F., H. J. Morton, and R. C. Parker. 1950. Nutrition of animal cells in tissue culture. I. Initial studies on a synthetic medium. Proc. Soc. Exp. Biol. Med. 73: 1–8.

    PubMed  CAS  Google Scholar 

  24. Vielkind, J., U. Vielkind, and F. Anders. 1973. DNA synthesis in genetically determined melanomas of platyfish-swordtail hybrids. Z. Krebsforsch. 80: 103–112.

    Article  CAS  Google Scholar 

  25. Giard, D. J., S. A. Aaronson, G. J. Todaro, P. Arnstein, J. H. Kersey, H. Dosik, and W. P. Parks. 1973. In vitro cultivation of human tumors: Establishment of cell lines derived from a series of solid tumors. J. Nat. Cancer Inst. 51: 1417–1423.

    PubMed  CAS  Google Scholar 

  26. Hu, F. 1971. Melanin production in mammalian cell cultures. In: T. Kawamura, T. B. Fitzpatrick, and M. Seiji (Eds.),Biology of Normal and Abnormal Melanocytes. University Park Press, Baltimore-London-Tokyo, pp. 303–328.

    Google Scholar 

  27. Ide, H. 1974. Proliferation of amphibian melanophores in vitro. Dev. Biol. 41: 380–384.

    Article  PubMed  CAS  Google Scholar 

  28. Kitano, Y., and F. Hu. 1970. Proliferation and differentiation of pigment cells in vitro. J. Invest. Dermatol. 55: 444–451.

    Article  PubMed  CAS  Google Scholar 

  29. Kreider, J. W., and M. E. Schmoyer. 1975. Spontaneous maturation and differentiation of B 16 melanoma cells in culture. J. Nat. Cancer Inst. 55: 641–647.

    PubMed  CAS  Google Scholar 

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Dedicated to Professor Dr. C. Kosswig on the occasion of his 75th birthday.

This work was supported by grants from the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 103, Marburg, Zellenergetik und Zelldifferenzierung), and is part of the thesis of C. K.

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Kuhn, C., Vielkind, U. & Anders, F. Cell cultures derived from embryos and melanoma of poeciliid fish. In Vitro 15, 537–544 (1979). https://doi.org/10.1007/BF02618156

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