Cultures of normal human mammary cells

Gaffney, Edwin V.; Polanowski, Frank P.; Blackburn, Susan E.; Lambiase, James T.; Burke, Robert E.

doi:10.1016/0045-6039(76)90001-4

Cited by 33 publications

(11 citation statements)

References 9 publications

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“…A better system seems to be primary culture, which is more closely related to the situation in vivo. Many investigators have contributed in the development of primary monolayer cultures of mammary cells (6,(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23). However, the major difficulty with primary mammary culture has been an inability to sustain continuous growth with a multifold increase in cell number characteristic of mammary cells in vivo during certain physiological and pathological states.…”

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confidence: 99%

“…However, the major difficulty with primary mammary culture has been an inability to sustain continuous growth with a multifold increase in cell number characteristic of mammary cells in vivo during certain physiological and pathological states. Therefore, the demonstration of mammary cell proliferation in primary monolayer culture has been limited primarily to [3H]thymidine incorporation (9)(10)(11)(12)(13)(14)(15). Recent cell culture studies (6,(16)(17)(18)(19)(20) have used cell counting to quantitate the effects of hormones and growth factors on the growth of mammary cells in primary monolayer culture.…”

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Sustained growth in primary culture of normal mammary epithelial cells embedded in collagen gels

Yang

Richards

Guzman

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

143

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Normal mammary epithelial cells from BALB/cfC3H middpregnant mice were freed from stromal cell types by Percoll density gradient centrifugation after collagenase digestion and were then embedded within collagen gels. Sustained growth leading to an increase in cell number was accomplished in response to cholera toxin and high concentrations of horse serum. The extent of growth was found to be dependent on the horse serum concentration, the maximum growth being attained at 50%. A serum concentration of 12.5% horse serum and 2.5% fetal calf serum, along with cholera toxin at 0.01 Lg/ml, allowed maintenance but failed to cause any significant increase in cell number during the experimental period of 2 weeks. This same maintenance medium was used to determine the effects of various exogenously added steroids, protein hormones, and organ extracts on the proliferation of mammary epithelial cells in culture. Hormones failed to elicit any proliferative response, but extracts of kidney, brain, uterus, and spleen produced proliferative responses equal to or greater than the response obtained with 50% horse serum and cholera toxin. Kidney extracts prepared from midpregnant mice, virgin mice, and virgin mice given pituitary isografts all showed comparable activities, suggesting that the concentration of stimulatory factor(s) was not influenced by the hormonal status of the donor. Normal mammary epithelial cells that had undergone a 10-to 15-fold increase in cell number over initial values during 2-3 weeks in culture were passaged to secondary gel cultures. Outgrowths similar to those seen in primary culture were seen again in secondary culture. The present system provides a method for sustaining growth in culture of primary mammary epithelial cells from normal tissues.Classical endocrinology involving organ ablation and hormone therapy has developed the concept that pituitary, ovarian, and adrenocortical hormones are involved in mammary gland development (1, 2). Detailed analysis, however, requires an in vitro system in which the direct mitogenic effect of hormones can be assessed on appropriate target cells for prolonged periods. Early studies using organ and fragment cultures have shown insulin and growth factors to be the major factors responsible for mammary cell proliferation, but not the in vivo mammogenic hormones, an observation presenting a paradox with the in vivo situation (1, 2). Even insulin and the growth factors have been shown to cause only one or two rounds of DNA synthesis in vitro (3-6) and never a sustained growth.Recent studies indicate that, while estrogens have a direct mitogenic effect on a human mammary tumor cell line (MCF-7) (7) as assessed by an increase in cell number, estrogen mitogenicity cannot be demonstrated on the MTW9/PL mammary cell line (8). It has been proposed for the latter case that estrogen acts indirectly on the target cells through the production of intermediary growth factors that in turn promote growth of the mammary cell line (18). An inherent limitation of the use of e...

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Sustained growth in primary culture of normal mammary epithelial cells embedded in collagen gels

Yang

Richards

Guzman

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

143

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“…Philadelphia. Pa. (6). Line BT-20 (human breast carcinoma) developed and originally provided by Dr. E. Lasfargues.…”

Section: Methodsmentioning

confidence: 99%

Nude Mice of Different Strains in Parabiosis for Prolonged Periods of Time

Giovanella¹,

Hunter²,

Gary³

et al. 1984

Pathobiology

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“…Digestion of tissues with commercial preparations of collagenase releases stromal cells from organized a gregates of epithelial cells embedded in collagen $asfargues, 1957), and has been used for the isolation of mammary gland fragments from rats (Richards and Nandi, 1978), mice (Yang et al, 1979) and humans (Gaffney et al, 1976). Many investigators have implied that most, if not all, of the cells obtained under these conditions are lining epithelial cells.…”

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confidence: 99%

“…Many biochemical investigations of normal human breast involving, for example, studies of the metabolism of carcinogenic polycyclic hydrocarbons, require routine production of relatively large numbers (lo8 to lo9) of viable epithelial cells, essentially free of stromal cells, from human breast tissue obtained from mastectomies or, preferably, reduction mammaplasties. Digestion of tissues with commercial preparations of collagenase releases stromal cells from organized a gregates of epithelial cells embedded in collagen $asfargues, 1957), and has been used for the isolation of mammary gland fragments from rats (Richards and Nandi, 1978), mice (Yang et al, 1979) and humans (Gaffney et al, 1976). Many investigators have implied that most, if not all, of the cells obtained under these conditions are lining epithelial cells.…”

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Preparation and identification of human breast epithelial cells in culture

Easty

Monaghan

et al. 1980

Intl Journal of Cancer

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The routine preparation of gram quantities of lobuloalveolar and ductal structures from human reduction mammaplasties by treatment with collagenase is described. When cultured on plastic or glass surfaces these structures give rise, initially, to epithelial sheets consisting of cells which retain many morphological characteristics of myoepithelial cells and do not stain with an antiserum which reacts with the surfaces of the lining epithelium of breast ducts and lobulo-alveoli. Subsequently, cells which resemble the lining epithelial cells migrate from these structures and react strongly with the antiserum. Cell proliferation in these cultures is minimal. Explants of major ducts dissected from mastectomies produce vigorously proliferating epithelial sheets which contain morphologically similar cells not identifiable as either lining epithelium or myoepithelium, although nearly all the cells in direct contact with the medium react with the lining epithelium specific antiserum.

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Cultures of normal human mammary cells

Cited by 33 publications

References 9 publications

Sustained growth in primary culture of normal mammary epithelial cells embedded in collagen gels

Sustained growth in primary culture of normal mammary epithelial cells embedded in collagen gels

Nude Mice of Different Strains in Parabiosis for Prolonged Periods of Time

Preparation and identification of human breast epithelial cells in culture

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