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  • 1
    Electronic Resource
    Electronic Resource
    Method for culturing mammary epithelial cells in a rat tail collagen gel matrix (1983)
    Springer
    Methods in cell science 8 (1983), S. 31-36 
    Details
    ISSN: 1573-0603
    Keywords: growth ; differentiation ; mammary epithelium ; collagen gel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mammary glands are enzymatically dissociated and the resulting tissue digest enriched for epithelial cells by isopycnic banding on a density gradient of Percoll. The cells are embedded within a rat tail collagen gel matrix and fed with the appropriate medium. Growth and differentiation are superior in such a system when compared to culture on plastic, using identical media.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01834632
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  • 2
    Electronic Resource
    Electronic Resource
    A two-step technique for the cryopreservation of normal mouse mammary tissue (1983)
    Springer
    Methods in cell science 8 (1983), S. 37-39 
    Details
    ISSN: 1573-0603
    Keywords: two-step ; cryopreservation ; mammary tissue ; DMSO
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Dimethylsulfoxide is used as a cryoprotectant for the preservation of mammary tissue to be used in primary cell culture. The method involves a two-step process that reduces the sample temperature to −20° C for a short-time, which is followed by storage at −90° C or immersion into liquid nitrogen. Comparison between fresh and frozen-thawed tissue was made by the cells' ability to grow inside collagen gel in response to various hormones and growth factors. Growth of frozen-thawed and collagenase-digested tissue achieved 70 to 90% the DNA values of their nonfrozen counterparts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01834633
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  • 3
    Electronic Resource
    Electronic Resource
    Collagen gel culture system and analysis of estrogen effects on mammary carcinogenesis (1984)
    Springer
    Archives of toxicology 55 (1984), S. 91-96 
    Details
    ISSN: 1432-0738
    Keywords: Hormones ; Mitogen ; Mammary gland ; Collagen gel culture ; Estrogen ; Cell proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The results obtained to date from studies dealing with the role of hormones, including estrogen, on growth of mammary epithelial cells inside the collagen gel are described. The collagen gel matrix culture system appears to be a suitable system to obtain in vivo-like effects of hormones on mammary cell in vitro. The results thus far indicate that prolactin along with progesterone or cortisol can stimulate mammary cell proliferation. Thus far, estrogen has not been found to be mitogenic in our in vitro system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00346045
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  • 4
    Electronic Resource
    Electronic Resource
    In situ localization of prolactin receptor message in the mammary glands of pituitaryisografted mice (1994)
    Springer
    Molecular and cellular biochemistry 132 (1994), S. 145-149 
    Details
    ISSN: 1573-4919
    Keywords: prolactin receptor ; mouse mammary gland ; in situ localization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The receptors for prolactin (PRLR) are expressed in many tissues including the mammary gland, a classical target tissue for prolactin (PRL), but the cellular localization of expression of the PRLR gene in mammary gland has not yet been identified. PRL is known to up regulate its own receptor. We therefore employed the pituitary isografted mouse as a model to distinguish the cells expressing PRLR since PRL blood levels are known to be constitutively elevated in these animals. Mammary glands of virgin or pituitary isografted mice were analyzed by northern blot orin situ hybridization with a digoxygenin-labeled cRNA probe. Northern analysis revealed the expression of 1.3 kb and 2.5 kb forms of PRLR corroborating the results of various laboratories studying other tissues. PRLR was barely detectable byin situ hybridization in non-isografted mice. Two weeks after pituitary isografting, however, PRLR expression was substantially increased in the epithelial cells of mammary ducts and alveoli. No signal was ever detected in the mammary stromal compartment of either virgin or pituitary-isografted mice. The localization of PRL-responsive cells to the parenchyma of the mammary gland suggests that epithelial cells are the mediators of PRL action and that the transcriptional regulation of PRLR expression by PRL is direct in the epithelial cell.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00926923
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  • 5
    Electronic Resource
    Electronic Resource
    The use of thioesterase II as a rat mammary epithelial cell-specific marker (1983)
    Springer
    Cell & tissue research 234 (1983), S. 57-70 
    Details
    ISSN: 1432-0878
    Keywords: Mammary ; Epithelial ; Thioesterase II ; Cell marker
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The avidin-biotin-peroxidase complex immunoperoxidase technique was employed to determine the intercellular distribution of thioesterase II in rat mammary glands. This enzyme is responsible for shifting the product specificity of the fatty-acid synthetase enzyme complex from long to medium chain fatty acids. Thioesterase II was found exclusively in the cells lining the lumen of the ductal and alveolar structures in glands from mature virgin (150 days old) and pregnant rats. The ductal cell staining intensity was considerably less than that of the alveolar cells in the mature virgin rat glands. No immunoreactive thioesterase II was found in the stromal, adipose, vascular, or myoepithelial components of the gland in the developmental stages examined. In the glands from immature virgin rats (40–45 days old) thioesterase II was again found only in the epithelial cells lining the lumen of the ductal and end-bud structures although this layer was usually more than one cell thick. Quantitative determination of thioesterase II activity in cytosol preparations revealed similar levels in mammary fragments from enzymatically-dissociated glands obtained from mature virgins and in end buds derived from immature virgins, but somewhat higher levels in mammary structures derived from late-pregnant animals. These immunohistological and biochemical results demonstrate thioesterase II's usefulness as a mammary epithelial cell-specific marker.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00217402
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  • 6
    Electronic Resource
    Electronic Resource
    Human breast epithelial cells in serum-free collagen gel primary culture: Growth, morphological, and immunocytochemical analysis (1987)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 133 (1987), S. 228-234 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Human breast epithelial cells derived from various sources (fibroadenoma, reduction mammoplasty, and mastectomy tissues from premenopausal patients) have been cultured in collagen gel matrix using serum-free medium. Response to various additives has been analyzed for growth-promoting effect when added to a basal medium containing insulin, cholera toxin, and BSA. A consistent observation has been the effect of EGF and cortisol in growth stimulation of human breast epithelial cells, while separately, each additive elicited only a small response. Under this condition, employing EGF and cortisol combinations, these cells gave rise to organized colonies consisting of clusters of cells, usually spherical, without any duct-like extensions. Ultrastructural and immunocytochemical studies, using a panel of monoclonal and polyclonal antibodies, have shown that cell types and features that can be identified in the original breast tissue can also be delineated in the progeny populations. The topographical feature, consisting of lumina surrounded by a single inner layer of epithelial cells and an outer layer of basal/myoepithelial cells, can be re-created in the collagen gel system starting from small clumps of cells.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041330205
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  • 7
    Electronic Resource
    Electronic Resource
    Growth stimulation by PGE2 and EGF activates cyclic AMP-dependent and -independent pathways in primary cultures of mouse mammary epithelial cells (1988)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 135 (1988), S. 509-515 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mammary epithelial cells from virgin Balb/c mice were isolated by collagenase digestion and cultured within collagen gels in serum-free basal medium containing insulin (10 μg/ml). Previous work has shown that linoleate or its metabolite, prostglandin E2 (PGE2), stimulate the growth of these cells only in the presence of a growth stimulant such as epidermal growth factor (EGF). Since PGE2 can stimulate cyclic AMP (cAMP) production, the role of cAMP in linoleate and EGF-stimulated growth was examined. The cAMP phosphodiesterase inhibitor, IBMX (0.1 mM), was found to augment growth when cells were cultured in the presence of both EGF and linoleate or PGE2, but not either factor alone. These results indicated that EGF does not stimulate proliferation via cyclic AMP mediated events but could synergize with cAMP events if cAMP levels were elevated by PGE2. When assayed in cells plated on top of collagen-coated culture dishes, cellular cyclic AMP levels were stimulated by PGE2, but only marginally by EGF. Although the stimulation of endogenous cAMP by PGE2 and IBMX was insufficient to stimulate growth in the absence of EGF, exogenous dibutyryl-cAMP (〉100 μg/ml) was able to do so showing that a sustained, and high level of cAMP (〉100 μg/ml) could stimulate growth in insulin-containing basal medium. EGF was capable of enhancing the cellular sensitivity to dibutyryl-cAMP but the converse was not observed. cAMP stimulation of growth was dependent upon a superphysiological concentration of insulin (10 μg/ml) or a physiological concentration of somatomedin-C. These results indicate that the proliferation of mouse mammary epithelial cells can be stimulated separately or in synergism by cAMP-dependent or -independent events.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041350320
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  • 8
    Electronic Resource
    Electronic Resource
    Growth effect of lithium on mouse mammary epithelial cells in serum-free collagen gel culture (1983)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 117 (1983), S. 290-296 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effect of lithium on the growth of mammary epithelial cells from adult virgin and midpregnant BALB/c or BALB/cfC3H mice was tested in a serum-free collagen gel culture system. The serum-free medium consisted of a 1:1 mixture of Ham's F12 and Dulbecco's Modified Eagle's medium supplemented with insulin, transferrin, cholera toxin, epidermal growth factor (EGF), and bovine serum albumin fraction V (BSA V). A multifold increase in cell number occurred during 10-12 days of culture in this medium. In dose-response studies in which the concentration of each component of this serum-free medium was varied in turn, the addition of LiCL (10 mM) enhanced growth at most concentrations of each factor. However, LiCL could not enhance growth in the absence of insulin or BSA V, but could replace EGF. The optimal concentration of LiCl was 5-10 mM; higher concentrations (20-80 mM) were toxic. KCl (1-10 mM) when added to the serum-free medium slightly stimulated growth; the addition of NaCl to the medium had little effect on growth. LiCl did not enhance the growth of cells from spontaneous mammary tumors of BALB/cfC3H mice.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041170303
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  • 9
    Electronic Resource
    Electronic Resource
    Differential response of normal rat mammary epithelial cells to mammogenic hormones and EGF (1985)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 125 (1985), S. 182-191 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A simple dissociation procedure and the collagen gel culture system have been utilized to determine the effects of mammogenic hormones and epidermal growth factor (EGF) on the proliferation of normal rat mammary epithelial (RME) cells in serum-free culture. Epithelial fragments, isolated from normal virgin F344 rat mammary glands by enzyme digestion followed by Percoll density gradient centrifugation, were embedded within a rat tail collagen matrix. A three-to four-fold increase in cell number was observed when ovine prolactin (PRL) and progesterone (P) were present in the basal medium during 7 days of culture. Mouse EGF stimulated one cell doubling during the same culture period.Isolated mammary organoids produced a ‘stellate’ type colony when PRL + P were present in the culture medium. These colonies were composed of small, tightly packed cuboidal cells. The addition of EGF to the basal medium produced a diffuse ‘basket’ type colony which was composed of large, elongate cells. When the complete hormonal and growth factor combination (PRL + P + EGF) was present, a ‘mixed’ type colony was observed which contained both the large and small epithelial cell types.Immunocytochemical analysis revealed that both the cuboidal and elongate cells present in the two colony types stained with antibodies to keratin indicating that these cells were epithelial in nature. The small cuboidal cells also expressed thioesterase II and alpha-lactalbumin, both specific for secretory mammary epithelial cells. The large, elongate cell type, however, was positive for actin but did not stain for either secretory epithelial specific marker. The results reported here suggest that normal rat mammary tissue may contain two epithelial populations, one which responds to PRL + P and the other which responds to EGF.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041250203
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  • 10
    Electronic Resource
    Electronic Resource
    Multifunctional phosphatidic acid signaling in mammary epithelial cells: Stimulation of phosphoinositide hydrolysis and conversion to diglyceride (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 163 (1995), S. 561-569 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have shown previously that phosphatidic acid esterified to polyunsaturated fatty acids is mitogenic for primary cultures of mouse mammary epithelial cells embedded within collagen gels. We hypothesized that this mitogenic competence resulted from the ability of this phospholipid to activate multiple signal transduction pathways in mammary epithelium. A closer examination of this hypothesis was undertaken by examining the effect of exogenous phosphatidic acid on phosphoinositide (PI) hydrolysis and its intracellular metabolism to diglyceride, an activator of protein kinase C. For assays of phosphoinositide-specific phospholipase C activation, mammary epithelial cells from virgin Balb/c mice were isolated by collagenase dissociation of mammary glands and cultured on the surface of Type I collagen-coated culture dishes. Phosphatidic acid (PA) stimulated a sustained increase in inositol phosphates and caused inositol phospholipid depletion when added to cells in which inositol phospholipids were prelabeled with 3H-myoinositol. This effect was specific for PA among phospholipids tested. Neither lineoleic acid, that can be released from PA, nor prostaglandin E2 affected PI hydrolysis. When mammary epithelial cells were cultured inside collagen gels in the presence of exogenous PA or phosphatidylcholine (PC) radiolabeled with 3H-glycerol, PA was found to persist intracellularly and be dephosphorylated to diglyceride (an activator of protein kinase C) to a greater extent than PC, a nonmitogenic phospholipid. In contrast to PA, epidermal growth factor (EGF) only slightly stimulated PI hydrolysis, showing that these two different growth-promoting factors do not actively couple to the same signal transduction pathways in mammary epithelial cells. These results show that PA may activate multiple pathways in mammary epithelial cells either directly or via its metabolism to diglyceride. © 1995 Wiley-Liss, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041630317
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