[37] Techniques for studying development of normal mammary epithelial cells in organ culture

Topper, R J; Oka, Takami; Vonderhaar, BK

doi:10.1016/s0076-6879(75)39039-3

Cited by 96 publications

(38 citation statements)

References 36 publications

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“…The extent of morphological and functional differentiation exhibited by the control mammary glands in our experiments (Figures 2, 5, and 7) are similar to previous reports (Terry et al, 1977;Mehta et al, 1980;Ganguly et al, 1981;Binas et al, 1992;Plaut et al, 1993;Li et al, 1995). In addition to proliferation and differentiation, mammary glands in whole organ culture can be induced to involute, a process mediated by apoptosis (Atwood et al, 1995;Casey et al, 1996), by the removal of the lactogenic hormones and culturing only with insulin (Topper et al, 1975). In contrast to the use of an involution-inducing medium, severe apoptosis of epithelial cells in the differentiation medium is achieved by vanadate treatment.…”

Section: Discussion Whole Organ Culture To Study Mammary Gland Develosupporting

confidence: 91%

“…The primed control mammary glands were fixed immediately after explantation for the determination of preculture conditions. The collected glands were transferred to siliconized lens paper (Topper et al, 1975) and then floated on the culture medium (2 ml per well) in a 6-well plate (Costar Corporation, Cambridge, MA). The culture medium was Waymouth's medium 752/1 supplemented with penicillin (100 U/ml), streptomycin (100 g/ml), insulin (5 , aldosterone (0.1 , hydrocortisone (0.1 , and prolactin (1 g/ml).…”

Section: Whole Organ Culture (Woc)mentioning

confidence: 99%

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Vanadate disrupts mammary gland development in whole organ culture

Gallo-Hendrikx

Murray

Vonderhaar

et al. 2001

Developmental Dynamics

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Protein tyrosine kinases and phosphatases are signaling molecules involved in all aspects of development, including proliferation, differentiation, and apoptosis. How disruption of protein tyrosine phosphatase affects mammary gland development is not entirely clear. We examined the effects of sodium vanadate, which is known to primarily inhibit tyrosine phosphatases, in mouse mammary gland development in whole organ culture. Mammary epithelial differentiation was effectively inhibited by vanadate in a dosedependent manner as indicated by lack of epithelial alveoli compared to the contralateral nontreated gland controls. Mammary glands in the differentiation medium after four days in the presence of vanadate did not differentiate into alveoli. Instead, they exhibited prominent terminal end buds and lost the distinctive epithelial structures. The inhibitory effect of vanadate on mammary epithelial cell differentiation was irreversible after one day of treatment. Immunohistochemical staining for PCNA (Proliferating Cell Nuclear Antigen) showed that vanadate-treated glands exhibited elevated proliferation signals in the differentiation medium. Expression of ␤-casein protein in the vanadate-treated glands decreased dramatically and progressively. Short-term exposure (up to 72 hours) of mammary glands to vanadate resulted in an increase in mammary epithelial cell density and loss of organization of the mammary structures. TUNEL assay of mammary glands with prolonged exposure to vanadate revealed widespread apoptosis. Furthermore, some cells were still proliferating or expressing ␤-casein after prolonged exposure to vanadate. Taken together, these data indicate that vanadate treatment blocks mammary epithelial cell differentiation and promotes abnormal proliferation and apoptosis, likely through the inhibition of protein tyrosine phosphatase-mediated signaling.

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Section: Discussion Whole Organ Culture To Study Mammary Gland Develosupporting

confidence: 91%

Section: Whole Organ Culture (Woc)mentioning

confidence: 99%

Vanadate disrupts mammary gland development in whole organ culture

Gallo-Hendrikx

Murray

Vonderhaar

et al. 2001

Developmental Dynamics

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“…28,29 In the presence of insulin, prolactin, and hydrocortisone, cultured mammary explants remain in a lactating state. 30 After the lactation period the gland undergoes an extensive remodeling process that leads to the involution of the epithelial structure until a state is reached resembling that of a virgin gland. 28,31 The involution process is associated with changes in the pattern of gene expression, with apoptosis of about 80% of the epithelial cells and with the production and activity of extracellular matrix degrading proteases.…”

Section: Induction Of Apoptosismentioning

confidence: 99%

Physiological apoptosis in hormone-dependent tissues: involvement of caspases

et al. 1999

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Physiological apoptosis in mammals is a type of programmed cell death, an important element in the developmental repertoire ensuring tissue homeostasis and proper disposal of cells that are no longer needed, such as milk-producing epithelial cells in the mammary gland after lactation, luteal cells in the post partum Corpus luteum or secretory cells in the prostate after castration. Although incompletely described, apoptosis in hormone-dependent tissues is apparently initiated and executed using common biochemical strategies. These include survival pathways governed by local and systemic factors and hormones, diverse regulatory pathways and caspase-dependent execution pathways. Using an antibody that recognizes processed effector caspases or a fluorogenic caspase substrate, we present for the first time evidence that caspases are activated in the mammary gland, in the prostate and in the ovary at the time when apoptosis occurs. Most likely phagocytosis of apoptotic cells by neighboring cells may represent an important step, since only a modest involvement of professional phagocytes is apparent. Here, we will summarize and discuss recent data and will attempt to draw a generalized picture of how physiological apoptosis may occur in these organs.

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“…* To whom correspondence should be addressed Mammary gland explants were prepared from abdominal glands and cultured as in [6]. The amount of a-lactalbumin in the cultured tissue was determined by a modified method of [7], using pure bovine o-lactalbumin as the standard [8].…”

Section: Introduction 2 Materials and Methodsmentioning

confidence: 99%