Skip to main content
Cytotechnology logoLink to Cytotechnology
. 1998 Mar;26(2):83–90. doi: 10.1023/A:1007911619537

Isolation, culture and characterization of a primary fibroblast cell line from channel catfish

Quiyang Zhang 1, Richard K Cooper 2, William R Wolters 3, Terrence R Tiersch 1
PMCID: PMC3466682  PMID: 22358545

Abstract

A primary cell line (designated as CCf) derived from caudal fin tissue of channel catfish, Ictalurus punctatus, was developed using explant techniques. The cell line grew fastest in media supplied with FBS and channel catfish serum. The duplication time of the cell line under optimal conditions was ∼56 h at a plating density of 1.1 × 105 cells/ml. The cell line has been propagated continuously for 25 passages (1:4 dilution per passage), cryopreserved, and recovered successfully at different passages. The cultured cells had fibroblastic morphology, and synthesized fibronectin and Type I and III collagens in the cytoplasm. The cell line maintained the normal diploid chromosome number (58) of channel catfish throughout the experiment. Nucleolus organizer regions were located on the short arms of a pair of medium-sized submetacentrics, which is typical for channel catfish. This study provides a method for acquiring a cell line from juvenile catfish without sacrifice, and is especially useful for early screening of valuable fishes.

Keywords: fibroblast cell line, characterization, Ictalurus punctatus, teleost

Full Text

The Full Text of this article is available as a PDF (126.2 KB).

References

  1. Amemiya CT, Bickham JW and Gold JR (1984) A cell culture technique for chromosome preparation in cyprinid fishes. Copeia 1984: 232-235.
  2. Avella M, Berhaut J, Payan P. Primary cuture of gill epithelial cells from the sea bass Dicentrarchus labrax. In Vitro Cell. Dev. Biol. 1994;30A:41–49. doi: 10.1007/BF02631417. [DOI] [PubMed] [Google Scholar]
  3. Bouchard B, Fuller BB, Vijayasaradhi S, Houghton AN. Induction of pigmentation in mouse fibroblasts by expression of human tyrosinase cDNA. J. Exp. Med. 1989;169:2029–2042. doi: 10.1084/jem.169.6.2029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bowser PR, Plumb JA. Fish cell lines: establishment of a line from ovaries of channel catfish. In Vitro Cell. Dev. Biol. 1980;16:363–368. doi: 10.1007/BF02618357. [DOI] [PubMed] [Google Scholar]
  5. Freshney RI. Culture of Animal Cells: a Manual of Basic Technique. 3rd ed. New York: Wiley Liss; 1994. [Google Scholar]
  6. Fryer JL, Lannan CN. Three decades of fish cell culture: a current listing of cell lines derived from fishes. J. Tissue Cult. Methods. 1994;16:87–94. doi: 10.1007/BF01404816. [DOI] [Google Scholar]
  7. Gay S, Matin GR, Muller PK, Timple R, Kuhn K. Simultaneous synthesis of type I and III collagen by fibroblasts in culture. Proc. Natl. Acad. Sci. USA. 1976;73:4037–4040. doi: 10.1073/pnas.73.11.4037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Howel WM, Black DA. Controlled silver staining of nucleolus organizer regions with a protective colloidal developer: a one step method. Experientia. 1980;36:1014–1015. doi: 10.1007/BF01953855. [DOI] [PubMed] [Google Scholar]
  9. Hynes R, Yamada KM. Fibronectins: Multifuctional modular glycoproteins. J. Cell Biol. 1982;95:369–377. doi: 10.1083/jcb.95.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kang J, Caprio J. In vivoresponses of single olfactory receptor neurons in the channel catfish. Ictalurus punctatus. J. Neurophysiol. 1995;73:172–177. doi: 10.1152/jn.1995.73.1.172. [DOI] [PubMed] [Google Scholar]
  11. LeGrande WH, Dunham RA and Smitherman RO (1984) Karyology of three species of catfishes (Ictaluridae: Ictalurus) and four hybrid combinations. Copeia 1984: 873-878.
  12. Miller NW and McKinney EC (1994) In vitroculture of fish leukocytes. In: Hochachka and Mommensen (eds) Biochemistry and Molecular Biology of Fishes (pp. 339-351). Elsevier Science B.V.
  13. Miller NW, Chinchar VG, Clem LW. Development of leukocyte cell lines fromthe channel catfish (Ictalurus punctatus) J. Tissue Cult. Methods. 1994;16:117–123. doi: 10.1007/BF01404820. [DOI] [Google Scholar]
  14. Miller NW, Rycyzyn MA, Wilson MR, Warr GW, Naftel JP, Clem LW. Development and characterization of channel catfish long term B cell lines. J. Immunol. 1994;152:2180–2189. [PubMed] [Google Scholar]
  15. Noga EJ. Establishment of primary cell cultures using short-term trypsinization of organ fragments. J. Tissue Cult. Methods. 1980;6:55–56. doi: 10.1007/BF01666016. [DOI] [Google Scholar]
  16. Porter KR, Pappas GD. Collagen formation by fibroblasts of the chick embryo dermis. J. Biophys. Biochem. Cytol. 1959;5:153–166. doi: 10.1083/jcb.5.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stickney RR. Culture of Non-Salmonid Fishes. Boca Raton: CRC Press; 1993. [Google Scholar]
  18. Thune RL, Stanley LA and Cooper RK(1993) Pathogenesis of warm-water bacterial fish pathogens. Ann. Rev. Fish Diseases, 37-68.
  19. Vallejo AN, Ellsaesser CF, Miller NW, Clem LW. Spontaneous development of functionally active long-term monocyte-like cell lines from channel catfish. In Vitro Cell. Dev. Biol. 1991;27A:279–286. doi: 10.1007/BF02630904. [DOI] [PubMed] [Google Scholar]
  20. Wise GE, Lin F, Fan W. Culture and characterization of dental follicle cells from rat molars. Cell Tissue Res. 1992;267:483–492. doi: 10.1007/BF00319370. [DOI] [PubMed] [Google Scholar]
  21. Wohlschlag N, delos Snatos L, Dehn PF. Preparation of primary cultures of adult and juvenile catfish (Ictalurus punctatus) hepatocytes. J. Tissue Cult. Methods. 1989;12:53–55. doi: 10.1007/BF01404642. [DOI] [Google Scholar]
  22. Wolters WR. Channel catfish breeding and selection programs: Constraints and future prospects. In: Main KL, Reynolds B., editors. Selective Breeding of Fishes in Asia and the United States. Honolulu: The Oceanic Institute; 1993. pp. 82–95. [Google Scholar]
  23. Wolters WR, Chrisman CL and Libey GS (1981). Lymphocyte culture for chromosomal analysis of channel catfish, Ictalurus puntatus. Copeia 1981: 503-504.
  24. Yamada KM. Immunological characterization of a major transformation-sensitive fibroblast cell surface glycoprotein: Localization, redistribution, and role in cell shape. J. Cell Biol. 1978;78:520–541. doi: 10.1083/jcb.78.2.520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Zhang Q. Cytogenetic and molecular analysis of the channel catfish (Ictalurus punctatus) genome. Baton Rouge: Louisiana State University; 1996. [Google Scholar]
  26. Zhang Q and Tiersch TR (1997) Chromosomal inheritance patterns of intergeneric hybrids of ictalurid catfishes: Odd diploid numbers with equal parental contributions. J. Fish Biol. In press. [DOI] [PubMed]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.

RESOURCES