, Volume 32, Issue 1, pp 1–7 | Cite as

Obtaining cell proliferation for chromosome preparation in gill tissue culture of the oyster Crassostrea gigas

  • Michel Cornet


The present results were obtained in the course of theadjustment to the oyster Crassostrea gigas of atissue culture technique recently developed for themussel Mytilus edulis. With respect to theprotocol originally described, the effects of twomodifications are reported: (1) replacement of chickembryo extract by chicken serum for medium enrichment,and (2) achievement of cultures in rotating tubes(roller drum) in place of stationary condition.Paradoxical results were obtained: whereas takenseparately, each modification exerted a negativeeffect which is statistically significant, combinated,they exerted a high positive effect representing athree-fold increase of the mean metaphase spreadnumber per slide (i.e. 71.5). Hypotheses are proposedto explain the mechanisms involved. It is suggestedthat the two additives work differently and thatcultures with chick embryo extract enriched mediumcould not withstand the condition generated by theroller drum. Conversely, cultures performed withchicken serum enriched medium would be in a betterphysiological state and the roller allow to obtain acell proliferation after only six days of incubation.

chromosome culture technique invertebrata medium composition mollusca tissue culture 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ahmed M and Sparks AM (1967) A preliminary study of chromosomes of two species of oysters (Ostrea lurida and Crassostrea gigas). J Fish Res Board Can 24: 2155-2159.Google Scholar
  2. Beaumont AR (1994) The application and relevance of genetics in aquaculture. In: Beaumont AR (ed) Genetics and Evolution of Aquatic Organisms. Chapman & Hall, London, pp. 467-486.Google Scholar
  3. Brewster F and Nicholson BL (1979) In vitro maintenance of amoebocytes from the American oyster Crassostrea virginica. J Fish Res Board Can 36: 461-467.Google Scholar
  4. Colombo L, Bortolussi M., Belvedere P and Battaglia B (1990) Genetic improvement in aquaculture. In: Hummel H (ed) The Genetics of Marine and Estuarine Organisms. DIHO, Yerseke NDL, pp. 32-33.Google Scholar
  5. Cornet M (1993) A short-term culture method for chromosome preparation from somatic tissues of adult mussel (Mytilus edulis). Experientia 49: 87-90.Google Scholar
  6. Cornet M (1995) A study of some factors affecting the efficiency of in vitro mussel tissue culture for chromosome preparation. Cytobios 84: 7-12.Google Scholar
  7. Domart-Coulon I, Doumenc D, Auzoux-Bordenave S and Le Fichan Y (1994) Identification of media supplements that improve the viability of primarily cell cultures of Crassostrea gigas oysters. Cytotechnology 16: 109-120.Google Scholar
  8. Griffiths JB (1990) Scale-up of suspension and anchorage-dependent animal cells. In: Pollard JW and Walker JM (eds) Methods in Molecular Biology, vol. 5, Animal Cell Culture. Humana Press, Clifton NJ, pp. 49-63.Google Scholar
  9. Hummel H and Patarnello T (1994) Genetic effects of polluants on marine and estuarine invertebrates. In: Beaumont AR (ed) Genetics and Evolution of Aquatic Organisms. Chapman & Hall, London, pp. 425-434.Google Scholar
  10. Ieyama H and Inaba I (1974) Chromosome numbers of ten species in four families of Pteriomorphia (Bivalvia). Venus 33: 129-137.Google Scholar
  11. Ieyama H, Takubo M and Moribe S (1979) Explant culture of the oyster, Crassostrea gigas. Mem Ehime Univ Sci, B, 8: 1-4.Google Scholar
  12. Jayme D, Watanabe T and Shimada T (1997) Basal medium development for serum-free culture: a historical perspective. Cytotechnology 23: 95-101.Google Scholar
  13. Jones BM (1966) Invertebrate tissue and organ culture in cell research. In: Willmer EN (ed) Cells and tissues in culture. Methods, biology and physiology, vol. 3. Academic Press, London, pp. 397-457.Google Scholar
  14. Kadouri A and Spier RE (1997) Some myths and messages concerning the batch and continuous culture of animal cells. Cytotechnology 24: 89-98.Google Scholar
  15. Kleinschuster SJ and Swink SL (1992) In vitro culture of presumptive nervous tissue of Crassostrea virginica (Gmelin, 1791). J Shellfish Res 11: 349-361.Google Scholar
  16. Le Deuff RM, Lipart C and Renault T (1994) Primary culture of Pacific oyster, Crassostrea gigas, heart cells. J Tissue Cult Meth 16: 67-72.Google Scholar
  17. Li MF, Stewart JE and Drinnan RE (1966) In vitro cultivation of cells of the oyster Crassostrea virginica. J Fish Res Board Can 23: 595-599.Google Scholar
  18. Macgregor HC and Varley JM (1983) Working with Animal Chromosomes. John Wiley & Sons, New york.Google Scholar
  19. Moore MN, Lowe DM, Livingstone DR and Dixon DR (1986) Molecular and cellular indices of polluant effects and their use in environmental impact assessment. Wat Sci Tech 18: 223-232.Google Scholar
  20. Ozturk SS (1996) Engineering challenges in high density cell culture systems. Cytotechnology 22: 3-16.Google Scholar
  21. Palomares LA and Ramirez OT (1996) The effect of dissolved oxygen tension and the utility of oxygen uptake rate in insect cell culture. Cytotechnology 22: 225-237.Google Scholar
  22. Perkins FA and Menzel RW (1964) Maintenance of oyster cells in vitro. Nature 204: 1106-1107.Google Scholar
  23. Penso G and Balducci D (1963) Tissue culture in biological research. Elsevier, Amsterdam.Google Scholar
  24. Renault T, Flaujac G and Le Deuff RM (1995) Isolation and culture of heart cells from the European flat oyster, Ostrea edulis. Meth Cell Sci 17: 199-205.Google Scholar
  25. Sami S, Ahmed I and Faisal M (1991) In vitro cultures of oyster Crassostrea virginica cells: stimulation by mitogens. Cheasapeake Res Cons Pub 137: 403-407.Google Scholar
  26. Sokal RR and Rohlf FJ (1981) Biometry. Freeman WH, San Francisco.Google Scholar
  27. Spier RE (1997) Factors limiting the commercial application of animal cells in culture. Cytotechnology 23: 113-117.Google Scholar
  28. Thiriot-Quiévreux C and Ayraud N (1982) Les caryotypes de quelques espèces de Bivalves et de Gastéropodes marins. Mar Biol 70: 165-172.Google Scholar
  29. Thiriot-Quiévreux C (1984) Analyse comparée des caryotypes d'Ostreidae (Bivalvia). Cah Biol Mar 25: 407-418.Google Scholar
  30. Thiriot-Quiévreux C, Noel T, Bougrier S and Dallot S (1988) Relationships between aneuploidy and growth rate in pair mating of the oyster Crassostrea gigas. Aquaculture 75: 89-96.Google Scholar
  31. Thiriot-Quiévreux C and Insua A (1992) Nucleolar organiser region variation in the chromosomes of three oyster species. J Exp Mar Biol Ecol 157: 33-40.Google Scholar
  32. Tripp MR, Bisignani LA and Kenny MT (1966) Oyster amoebocytes in vitro. J Invert Pathol 8: 137-140.Google Scholar
  33. Vago C and Chastang S (1960) Culture de tissus d'huître. C R Acad Sci Paris 250: 2751-2753.Google Scholar
  34. Wallis AM (1972) A new technique for the preparation of primary cultures of oyster tissue. Lab Pract 21: 32.Google Scholar
  35. Wen CM, Kou GH and Chen SN (1993) Establishment of cell lines from the Pacific oyster. In Vitro Cell. Dev Biol 29A: 901-903.Google Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Michel Cornet
    • 1
  1. 1.Laboratoire d'Océanographie Biologique, CNRS UMR 5805Université Bordeaux ITalence CedexFrance

Personalised recommendations