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Methods in Cell Science

, Volume 22, Issue 1, pp 33–41 | Cite as

Ten commandments for preventing contamination of primary cell cultures

  • Janet L. Vierck
  • Katherine Byrne
  • Priya S. Mir
  • Michael V. Dodson
Article

Abstract

Procedures for preventing contamination in primary cell cultures must be carefully defined and strictly followed in order to obtain healthy cells. Protocols have been developed and refined in our laboratory for establishing primary cultures of muscle and fat stem cells without contamination from a variety of animals. Contamination of cell cultures is not only frustrating, but is also very expensive both in time and loss of materials. Through the consistent use of proper aseptic techniques, most instances of contamination may be avoided. We suggest that the basic principles detailed here will find wide applicability in the culturing of primary cells without contamination from many different types of animals and tissues.

Aseptic technique Cell culture Contamination Sterilization Tissue collection 

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References

  1. 1.
    Adams RLP (1980). Cell culture for biochemists (Laboratory techniques in biochemistry and molecular biology, vol 8). Amsterdam, The Netherlands: Elsevier/North-Holland Biomedical Press.Google Scholar
  2. 2.
    Baker (1995). The Baker Company Catalogue. Sanford, ME: The Baker Company.Google Scholar
  3. 3.
    Corning (1981). General procedures for primary cell culture. Corning Incorporated: TC-14.Google Scholar
  4. 4.
    Corning (1987). General procedures for the cell culture laboratory. Corning Incorporated: TC-CGW-4A.Google Scholar
  5. 5.
    Corning (1989). General guide for identifying and correcting common cell culture growth and attachment problems. Corning Incorporated: TC-CI-27.Google Scholar
  6. 6.
    Dodson MV, McFarland DC, Martin EL, Brannon MA (1986). Isolation of satellite cells from ovine skeletal muscles. J of Tissue Cult Meth 10: 233-237.Google Scholar
  7. 7.
    Dodson MV, Martin EL, Brannon MA, Mathison BA, McFarland DC (1987). Optimization of bovine satellite cell-derived myotube formation in vitro. Tissue and Cell 19: 159-166.Google Scholar
  8. 8.
    Dodson MV, Mathison BA, Brannon MA, Martin EL, McFarland DC (1988). Comparison of ovine and rat muscle-derived satellite cells: Response to insulin. Tissue and Cell 20: 909-918.Google Scholar
  9. 9.
    Dodson MV, Mathison BA, Mathison BD (1990). Effects of medium and substratum on ovine satellite cell attachment, proliferation and differentiation in vitro. Cell Diff and Dev 29(1): 59-66.Google Scholar
  10. 10.
    Dodson MV, Vierck JL, Hossner KL, Byrne K and McNamara J (1997). The development and utility of a defined muscle and fat co-culture system. Tissue and Cell 29: 517-524.Google Scholar
  11. 11.
    Douglas WHJ, Dell'Orco RT (1979). Physical aspects of a tissue culture laboratory. In: Jakoby WB, Pastan IH (eds), Methods in Enzymology, vol. LVIII-cell culture, pp 3-18. New York: Academic Press.Google Scholar
  12. 12.
    Erickson S, Vierck J, Calkins D, Welter C, Krabbenhoft L, Byrne K, Greene E, Dodson MV (1998). Fusion of equine myogenic satellite cell strains in vitro. Oregon Sci J 14: 10-14.Google Scholar
  13. 13.
    Fisher Scientific Catalog (1998-99). Pittsburgh, PA: Fisher Scientific Company.Google Scholar
  14. 14.
    Freshney RI (1986). Introduction: Principles of sterile technique and cell propagation. In: Freshney RI (ed), Animal cell culture: A practical approach, pp 1-11. Oxford: IRL Press Limited.Google Scholar
  15. 15.
    Freshney RI (1994). Culture of animal cells. New York: Wiley-Liss.Google Scholar
  16. 16.
    GibcoBRL Products and Reference Guide (2000-2001). Grand Island, NY: Life Technologies.Google Scholar
  17. 17.
    Greene E, Raub R, Krabbenhoft L, Brannon B, Sullivan J, Hossner K, Dodson MV (1995). Initial observations on the growth factor regulation of equine satellite cells in vitro. Basic Appl Myol 5(1): 22-27.Google Scholar
  18. 18.
    Jeffrey JS (1997). Sanitation-Disinfection Basics. University of California: Cooperative Extension Service.Google Scholar
  19. 19.
    Knecht CD, Allen AR, Williams DJ, Johnson JH (1981). Fundamental Techniques in Veterinary Surgery. Philadelphia: W.B. Saunders Company.Google Scholar
  20. 20.
    McGarrity GJ (1979). Detection of contamination. In: Jakoby WB, Pastan IH (eds), Methods in enzymology, vol. LVIII-cell culture, pp 18-29. New York: Academic Press.Google Scholar
  21. 21.
    Mathison BD, Mathison BA, McNamara JP, Dodson MV (1989). IGF receptor analyses of satellite cellderived myotubes from two lines of Targhee rams selected for growth rate. Domest Anim Endocrin 6(3): 191-201.Google Scholar
  22. 22.
    Millipore Laboratory Catalogue, US Version (1997-98). Pittsburgh, PA: Fisher Scientific.Google Scholar
  23. 23.
    Molnar GR, Dodson MV (1992). Characterization of ovine semimembranosus muscle and associated satellite cells: Expression of fast and slow myosin heavy chain isoforms. Basic Appl Myol 2(3): 183-190.Google Scholar
  24. 24.
    Molnar GR, Dodson MV (1993). Satellite cells isolated from sheep muscle are heterogeneous. Basic Appl Myol 3(3): 173-180.Google Scholar
  25. 25.
    Nuaire (1996). Operation and Maintenance Manual-40 Labgard Laminar Flow Biological Safety Cabinet. Plymouth, MN: Nuaire, Inc.Google Scholar
  26. 26.
    Nuaire (1998). Operation and Maintenance Manual-Water-Jacketed US Autoflow Automatic CO2 Incubator. Plymouth, MN: Nuaire, Inc.Google Scholar
  27. 27.
    Perkins JJ (1969). Principles and methods of sterilization in health sciences. Illinois: Charles C. Thomas.Google Scholar
  28. 28.
    Powell Rl, Dodson MV and Cloud JG (1989). Cultivation and differentiation of satellite cells from skeletal muscle of the rainbow trout Salmo gairdneri. J Exptl Zool 250(3): 333-338.Google Scholar
  29. 29.
    Precision (1994). Installation and operation instructions for general purpose water baths. Chicago, IL: Precision Scientific, Inc.Google Scholar
  30. 30.
    Sigma (1999). Biochemicals and reagents for life science research. St. Louis, MO: Sigma-Aldrich Co.Google Scholar
  31. 31.
    Venkateswaran V, Brackett EJ, Vierck J, Cloud J, Dodson MV (1995). Substratum is an important determinant in growth factor regulation of troutderived satellite cells. Basic Appl Myol 5(3): 297-304.Google Scholar
  32. 32.
    Vierck J, McNamara J, Hossner K, Dodson MV (1995). Characterization of ovine skeletal muscle satellite cell strains in a defined medium formulated to enhance differentiation: Fusion and the IGF-I system. Basic Appl Myol 5(1): 12-21.Google Scholar
  33. 33.
    Vierck JL, McNamara JP, Dodson MV (1996a). Two alternative procedures to isolate adipofibroblasts from sheep skeletal muscle. Meth in Cell Sci 18(4): 309-314.Google Scholar
  34. 34.
    Vierck J, McNamara JP, Dodson MV (1996b). Proliferation and differentiation of progeny of ovine unilocular fat cells (adipofibroblasts). In Vitro Cell Dev Biol 32(10): 564-572.Google Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Janet L. Vierck
  • Katherine Byrne
  • Priya S. Mir
  • Michael V. Dodson

There are no affiliations available

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