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Methods in Cell Science

, Volume 17, Issue 4, pp 271–282 | Cite as

Use of microinjection to generate an immortalized bovine mammary cell line with both epithelial and myoepithelial characteristics

  • B. Zavizion
  • M. Van Duffelen
  • W. Schaeffer
  • I. Politis
Cell Biology

Abstract

Methods are described for the isolation, purification, and culture of bovine mammary epithelial cells. Individual cells were microinjected with a plasmid bearing the sequence for the simian virus 40 early region mutant tsA58 which encodes the thermolabile large T-antigen. After selection in G418-containing medium, surviving colonies were recovered and recloned. One of the clones, designated (E-TC), has been cultured for more than 300 population doublings and exhibits some very unusual properties. From low to middle confluency, E-TC cells exhibit the typical cobblestone morphology of epithelial cells. When E-TC cells remain in culture for 2 weeks, the majority of the cells in a single colony maintain the epithelial cobblestone-like morphology but cells at the periphery of the colony become larger, elongated, and less compact, a morphology more typical of myoepithelial-like cells. Myoepithelial cells, in co-culture, always surround epithelial cells (Zavizion et al., J. Dairy Sci. 75: 3381-3392). In co-culture with the BMM-UV bovine myoepithelial cells, E-TC cells are surrounded by the BMM-UV myoepithelial cells, thus the ET-C cells exhibit epithelial-like behavior. In co-culture with the MAC-T bovine epithelial cells, E-TC cells surround the MAC-T cells, thus the ET-C cells exhibit myoepithelial-like behavior. This suggests that E-TC cells may represent an immortalized bovine mammary stem cell (or precursor) which can exhibit myoepithelial-or epithelial-like properties depending upon certain culture conditions.

Key words

Bovine Cell culture Epithelial Mammary Microinjection Myoepithelial 

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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • B. Zavizion
    • 2
  • M. Van Duffelen
    • 2
  • W. Schaeffer
    • 1
  • I. Politis
    • 2
  1. 1.Department of Microbiology and Molecular GeneticsUniversity of VermontBurlingtonUSA
  2. 2.Department of Animal and Food Sciences, Terrill HallUniversity of VermontBurlingtonUSA

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