Veterinary Research Communications

, Volume 17, Issue 5, pp 341–351 | Cite as

An investigation of the suitability of three support matrices for the culture of cells derived from the secretory alveoli of the bovine mammary gland

  • W. G. F. Ditcham
  • A. W. Hill
  • A. P. Bland
  • J. A. Leigh


The suitability of three support matrices, (thick collagen gels, aluminium oxide and cellulose ester membranes, the latter two both thinly coated with collagen) for the production of primary cultures of bovine mammary epithelial cells was investigated. Single secretory alveoli were isolated from mammary tissue of animals in early lactation by enzymatic digestion and differential filtration. Cell growth was monitored by light and scanning electron microscopy. The cellulose ester membrane was found to give the best results, allowing growth of monolayers with a morphology closely resembling that of the natural epithelium of the gland. There were low levels of fibroblast contamination, and the membranes could be easily manipulated for further studies.


bacterial adhesion bovine cell culture epithelium mammary mastitis 



electron microscopy


N-2-hydroxyethylpiperazine-N′-2-ethanesulphonic acid


light microscopy


phase contrast microscopy


phosphate-buffered saline


Roswell Park Memorial Institute


scanning electron microscopy


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  1. Beachey, E.H., 1981. Bacterial adherence: Adhesin-receptor interactions mediating the attachment of bacteria to mucosal surfaces.Journal of Infectious Disease,143, 325–345Google Scholar
  2. Bissell, J.M., Glenn Hall, H. and Parry, G., 1982. How does the extracellular matrix direct gene expression?Journal of Theoretical Biology,99, 31–68Google Scholar
  3. Bramley, A.J. and Hogben, E.M., 1982. The adhesion of human and bovine isolates ofStreptococcus agalactiae (Group B) to bovine mammary gland epithelial cells.Journal of Comparative Pathology,92, 131–137Google Scholar
  4. Emerman, J.T. and Pitelka, D.R., 1977. Maintenance and induction of morphological differentiation in dissociated mammary epithelium on floating collagen membranes.In Vitro,99, 31–68Google Scholar
  5. Frost, A.J., 1975. Selective adhesion of microorganisms to the ductular epithelium of the bovine mammary gland.Infection and Immunity,12, 1154–1156Google Scholar
  6. Frost, A.J., Wanasinghe, D.D. and Woolcock, J.B., 1977. Some factors affecting selective adherence of microorganisms in the bovine mammary gland.Infection and Immunity,15, 245–253Google Scholar
  7. Frost, A.J., Hill, A.W. and Brooker, B.E., 1980. The early pathogenesis of bovine mastitis due toEscherichia coli.Proceedings of the Royal Society,209, 431–439Google Scholar
  8. Harper, M., Turvey, A. and Bramley, A.J., 1978. Adhesion of fimbriateEscherichia coli to bovine mammary-gland epithelial cellsin vitro.Journal of Medical Microbiology,11, 117–123Google Scholar
  9. Jones, G.W., 1977. The attachment of bacteria to the surfaces of animal cells. In: J.L. Reissig (ed.),Microbial Interactions. Receptors and Recognition, Series B, Vol. 3. (Chapman and Hall, London), 141–175.Google Scholar
  10. Lee, Y-H.L., Parry, G. and Bissell, M.J., 1984. Modulation of secreted proteins of mouse mammary epithelial cells by the collagenous substrata.Journal of Cell Biology,98, 146–155Google Scholar
  11. Mackenzie, D.D.S., Forsyth, I.A., Brooker, B.E. and Turvey, A., 1982. Culture of bovine mammary epithelial cells on collagen gels.Tissue and Cell,14, 231–241Google Scholar
  12. Millipore Technical Brief 016, 1986. Techniques to view, count and analyze cells grown onMillicell culture plate inserts. Millipore (UK) Ltd, The Boulevard, Blackmoor Lane, Watford, Herts, WD1 8YW, UKGoogle Scholar
  13. Nashar, T.O., Williams, M.R., Brown, P.J., Cripps, P.J. and Stokes, C.R., 1990. Fate and uptake of soluble and particulate antigens in the preparturient bovine mammary gland.Veterinary Immunology and Immunopathology,26, 125–141Google Scholar
  14. Ogawa, S.K., Yurberg, E.R., Hatcher, V.B., Levitt, M.A. and Lowy, F.D., 1985. Bacterial adherence to human endothelial cellsin vitro.Infection and Immunity,50, 218–224Google Scholar
  15. Opdebeek, J.P., Frost, A.J. and O'Boyle, D., 1988. Adhesion ofStaphylococcus aureus andEscherichia coli to bovine udder epithelial cells.Veterinary Microbiology,16, 77–86Google Scholar
  16. Park, C.S., Smith, J.J., Sasaki, M., Eigel, W.N. and Keenan, T.W., 1979. Isolation of functionally active acini from bovine mammary gland.Journal of Dairy Science,62, 537–545Google Scholar
  17. Parkkinen, J., Ristimaki, A. and Westerlund, B., 1989. Binding ofEscherichia coli S fimbriae to cultured human endothelial cells.Infection and Immunity,57, 2256–2259Google Scholar
  18. Richards, J., Larson, L., Yang, J., Guzman, R., Tomooka, Y., Osborn, R., Imagawa, W. and Nandi, S., 1983. Method for culturing mammary epithelial cells in a rat tail collagen matrix.Journal of Tissue Culture Methods,8, 31–36Google Scholar
  19. Sordillo, L.M., Oliver, S.P. and Akers, R.M., 1988. Culture of bovine mammary epithelial cells ind-valine modified medium: selective removal of contaminating fibroblasts.Cell Biology International Reports,12, 355–364Google Scholar
  20. Suard, Y.M.L., Haeuptle, M., Farinon, E. and Kraehenbuhl, J., 1983. Cell proliferation and milk protein gene expression in rabbit mammary cell cultures.Journal of Cell Biology,96, 1435–1442Google Scholar
  21. Tateyama, S., Furukawa, H., Yamaguchi, R. and Nosaka, D., 1990. Plate and collagen gel cultures of normal canine mammary epithelial cells.Research in Veterinary Science,49, 14–19Google Scholar
  22. Valentin-Weigand, P., Grulich-Henn, J., Chhatwal, G.S., Muller-Berghaus, G., Blobel, H. and Preissner, K.T., 1988. Mediation of adherence of streptococci to human endothelial cells by complement S protein (Vitronectin).Infection and Immunity,56, 2851–2855Google Scholar
  23. Wanasinghe, D.D., (1981). Adherence as a prerequisite for infection of the bovine mammary gland by bacteria.Acta Veterinaria Scandinavica,22, 109–117Google Scholar

Copyright information

© Kluwer Academic Publishers bv 1993

Authors and Affiliations

  • W. G. F. Ditcham
    • 1
  • A. W. Hill
    • 1
  • A. P. Bland
    • 1
  • J. A. Leigh
    • 1
  1. 1.Institute for Animal HealthCompton, NewburyUK

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