| Publication number |
CLPUB00665 |
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| Authors |
Huynh H.T. |
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| Title |
Establishment of bovine mammary epithelial cell lines: an in vitro model for lactation. |
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| Citation |
Thesis MSc (1990); McGill University Montreal; Montreal; Canada |
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| Web pages |
https://escholarship.mcgill.ca/concern/theses/2514nm65w |
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| Abstract |
Clonal cell lines were isolated from mammary gland tissue epithelial cell
cultures of lactating cows. Early passage clonal bovine mammary epithelial
cells (clone LMH17) gave rise to several established cell lines (MAC-T
lines) after being cotransfected with plasmids containing the temperature
sensitive mutant SV40 large T antigen gene (pBAPSV40TtsA58) and the
bacterial phosphotransferase gene (pSV2-neo). Unlike other cell types
which were transformed after being transfected with SV40, MAC-T cells
maintained many characteristics of non-transformed cells: MAC-T cells were
serum and anchorage dependent, showed contact inhibition, and were not
tumorigenic in immunodeficient mice. However, Southern transfer analysis
revealed an integrated SV40 gene and cells showed no senescence after 50
passages. These cells are morphologically indistinguishable from parental
LMH17 cells and retain the typical morphology of mammary epithelial cells.
Positive cytokeratin immunostaining and the absence of vimentin staining
indicated that these cells were epithelial in origin. MAC-T cells grew
rapidly on plastic substratum with a doubling time of approximately 17
hours and became differentiated when grown on floating collagen gels in
the presence of prolactin. The differentiated phenotype was characterized
to include (1) the ability to form secretory domes with a lumen from a
pavement of columnar cells; (2) increased casein mRNA abundance; (3)
increased alpha S and beta casein secretion; (4) increased number and size
of casein secretory vesicles; and (5) increased lactose synthesis and
secretion.
|
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| Cell lines |
CVCL_B3UP; LMH17 CVCL_U226; MAC-T |
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