Establishment and characterization of a caprine mammary epithelial cell line (CMEC) |
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Authors: | A G Pantschenko J Woodcock-Mitchell S L Bushmich T J Yang |
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Institution: | (1) Department of Pathobiology, University of Connecticut, 06269 Storrs, Connecticut;(2) Department of Molecular Physiology and Biophysics, College of Medicine, University of Vermont, 05405 Burlington, Vermont;(3) Department of Pathobiology, U-89, University of Connecticut, 06269-3089 Storrs, Connecticut |
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Abstract: | Summary We describe the establishment of a continuous, nontransformed cell line obtained from primary culture of a lactating (114
days postparturition) Anglo-Nubian (Capra hircus) goat mammary gland biopsy. These cells (CMEC), have been cultured in the presence of supraphysiologic concentrations of
insulin and hydrocortisone for more than 560 population doublings (over 80 passages) without any sign of senescence while
maintaining a normal/near-normal diploid chromosome modal number of 2n=60 and are responsive to contact inhibition of proliferation.
Cytoskeletal analysis indicates that CMECs are epithelial, without detectable fibroblastic or myoepithelial cells. When grown
at low density on plastic substratum, the cells tend to form island monolayer aggregates with the characteristics cobblestone
morphology of epithelial cells. With increasing density, the cells organize into lumen-like structures with various morphology
consisting of large and small vacuolized and nonvacuolized cells. Postconfluent cultures form epithelial raised dome-like
structures, implying a process of contact-induced differentiation. This is corroborated by positive immunocytochemistry to
lactation-specific proteins: β-casein and α-lactalbumin, which were predominantly expressed in dome-forming cells. We also
observed an overall modulation of cytokeratin 18/19 expression associated with number of days post subculture and with the
expression of lactation-specific proteins. Postconfluent cultures which contain lactation-specific, antibody-reactive, dome-like
structures showed a decreased expression of keratin 18 and no (null) expression for keratin 19. Lastly, cells cultured within
a collagen matrix show morphological differentiation with the organization of branching duct-like and acini-like structures.
This study suggests that CMECs are a useful in vitro model for study of mammary gland development and differentiation, in
particular, direct modulation of epithelial cells grown on plastic substratum or extracellular matrix without the influence
of stromal elements or the necessity and variability associated with primary cell culture or tissue explants. |
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Keywords: | differentiation dome-like structures lactogenesis cytoskeleton |
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