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Comparative analysis of casein synthesis during mammary cell differentiation in collagen and mammary gland development in vivo

Comparative analysis of casein synthesis during mammary cell differentiation in collagen and mammary gland development in vivo

Developmental Biology 109(2): 288-298

Substrata upon which epithelial cells are cultured modulate their morphology, growth, and ability to differentiate. Mouse mammary epithelial cells cannot be induced to synthesize caseins, a marker of cell differentiation, when grown on a plastic surface. An analysis was made of the effect of time within a collagen matrix on the ability of normal mammary epithelial cells to be induced to synthesize caseins and that response was compared to mammary gland development in vivo. Primary cultures of mammary cells from unprimed virgin BALB/c mice were embedded in rat-tail collagen gel mixtures and maintained in growth medium. Induction medium containing lactogenic hormones was added at various times. The cells were monitored every 3-7 days over a period of 8 wk for cell growth, casein synthesis, and ability to grow in vivo in cleared mammary fat pads. Casein accumulation was assayed quantitatively by an ELISA [enzyme-linked immunosorbent assay] competition assay and qualitatively by the immunoblot procedure using specific antisera prepared against purified mouse caseins. No marked differences in cell numbers and transplantability potential were observed among cells cultured for various times in collagen. Mammary cells grown in collagen for up to 8 wk retained the capacity to grow in vivo as normal ductal outgrowths. The duration of culture within collagen prior to hormonal stimulation did influence the kinetics of casein synthesis. Cells cultured for 1 wk in growth medium did not accumulate detectable levels of casein until after 3 wk of induction, whereas cells cultured for 2 or 4 wk responded by accumulating caseins after 2 wk and 3 days of induction, respectively. While the levels of total caseins that accumulated under optimal conditions of induction in culture approached levels found during lactation in vivo, the relative proportion of specific casein polypeptides synthesized in culture was altered from .alpha. casein (43K) in favor of the .beta. casein (30K) species. A period of culture within collagen is required to permit mammary epithelial cells to become responsive for hormone-induced differentiation. During growth within the collagen the cells synthesize and deposit extracellular matrix components important in modulating gene expression.

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Accession: 001320840

Download citation: RISBibTeXText

PMID: 3996752

DOI: 10.1016/0012-1606(85)90456-7

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