In vitro replication potential of serially passaged mammary parenchyma from mice with different reproductive histories

Choongkittaworn, Ngamchit; Hosick, Howard L.; Jones, Walis

doi:10.1016/0047-6374(87)90006-6

Cited by 9 publications

(2 citation statements)

References 22 publications

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“…5). The possibility cannot be completely excluded that the cells used in this study might change their character during culture (Choongkittaworn et al, 1987). However, we cultured the cells for no more than 3 days for purification and for preparation of a single cell suspension before soft agarose culture, so culture-induced abnormalities should be minimal (Daniel and DeOme, 1965).…”

Section: Discussionmentioning

confidence: 99%

“…The digest was passed through 250 pm and 48 pm Nitex mesh to separate epithelial organoids from larger gland fragments, single cells, and debris (Jones et al, 1983). The retentate on 48 pm mesh was pre-cultured in collagen gel to remove fibroblasts that migrate away from epithelium and into the gel (Choongkittaworn et al, 1987). Organoids were incubated for 2 days at 37°C in collagen gel matrix.…”

Section: Materials and Methods Animalsmentioning

confidence: 99%

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Transformed growth phenotype of mouse mammary epithelium in primary culture induced by specific fetal mesenchymes

Kanazawa

Hosick

1992

Journal Cellular Physiology

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When mesenchyme from fetal mammary or salivary gland is implanted into adult mouse mammary gland, adjacent epithelium responds with intense hyperplasia. The hyperplastic cells are more vulnerable than are non-stimulated cells to transformation in vivo by a chemical carcinogen or by mammary tumor virus. This system provides a potentially useful model for determining how stroma contributes to mammary tumorigenesis. We have developed co-culture systems and used them to investigate in more detail the nature of the signal produced by the mesenchyme cells. Monolayers of mesenchyme cells were prepared on tissue-culture wells. The mesenchyme cells were trapped on the surface by a thin overlay of agarose. Primary mammary epithelial cells were cultured atop this barrier layer, either as organoids in collagen gels for assessment of anchorage-dependent growth, or as single-cell dispersions in soft agarose for assessment of anchorage-independent growth. Our procedures for assay of anchorage-independent growth allow us for the first time to detect and measure this transformation-defining characteristic in non-immortalized mammary epithelial cells in primary culture. Fetal mammary fat pad precursor tissue and fetal salivary mesenchyme both stimulated anchorage-dependent growth of mammary epithelium, with cell number increasing as much as fifteenfold during a 6-day culture period. These same fetal tissues also stimulated anchorage-independent growth of the mammary epithelial cells, with colony-forming efficiencies of up to 40% in co-cultures with salivary mesenchyme. No colonies formed in the absence of mesenchyme. Cells of colonies contained keratin, which indicates that the colonies grew from epithelial cells and not from a contaminant of another cell type. When co-cultured epithelial cells were subsequently re-cultured in the absence of mesenchyme, they lost their ability to grow independent of anchorage. No colonies grew in co-cultures with fetal cells from heart, kidney, or lung, which is consistent with the lack of stimulation by these tissues in the mammary gland in vivo. A tumor promoter, 12-O-tetradecanoylphorbol acetate (TPA), also caused anchorage-independent growth of the dispersed mammary epithelial cells. Culture medium conditioned by primary or early-passage salivary mesenchyme cells was capable of stimulating growth under both anchorage-dependent and anchorage-independent conditions, confirming that these effects are mediated by a paracrine factor. The results indicate that stimulatory fetal mesenchymes produce soluble molecules that act analogously to transforming growth factors.

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Section: Discussionmentioning

confidence: 99%

Section: Materials and Methods Animalsmentioning

confidence: 99%

Transformed growth phenotype of mouse mammary epithelium in primary culture induced by specific fetal mesenchymes

Kanazawa

Hosick

1992

Journal Cellular Physiology

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Cell Culture Strategies for Analysis of Dietary Variables in Cancer

Hosick

1989

Carcinogenesis and Dietary Fat

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A co-culture system for studies of paracrine effects of stromal cells on the growth of epithelial cells

Kanazawa

Hosick

1992

Journal of Tissue Culture Methods

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SUMMARY:We have developed a co-culture system to study paracrine effects of fetal mesenchyme cells on the growth of primary mammary epithelial cells of the mouse. The method should be adaptable for study of interaction of a variety of cell populations. The procedure is simple and inexpensive. The culture consists of four layers: a monolayer of mesenchyme cells on a plastic culture dish, a soft agarose layer overlaying the cell monolayer, epithelial cells suspended in collagen gel placed atop the agarose, and culture medium above the collagen gel. The agarose layer prevents direct contact of two different cell populations but allows soluble molecules produced by either cell population to diffuse through the system, so as to contact and interact with the other cell population.

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In vitro replication potential of serially passaged mammary parenchyma from mice with different reproductive histories

Cited by 9 publications

References 22 publications

Transformed growth phenotype of mouse mammary epithelium in primary culture induced by specific fetal mesenchymes

Transformed growth phenotype of mouse mammary epithelium in primary culture induced by specific fetal mesenchymes

Cell Culture Strategies for Analysis of Dietary Variables in Cancer

A co-culture system for studies of paracrine effects of stromal cells on the growth of epithelial cells

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