Serum-free growth of normal and tumor mouse mammary epithelial cells in primary culture.

Imagawa, Walter; Tomooka, Yasuhiro; Nandi, Satyabrata

doi:10.1073/pnas.79.13.4074

Cited by 106 publications

(40 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The use of primary mammary cell cultures for the analysis of cell growth has been limited to studies of [aH]thymidine incorporation in short-term cultures, because of the inability to maintain mammary cells in long-term cultures. Due to recent developments in the cultivation of mammary cells using stromal collagen matrices (45), it became possible to maintain sustained growth and thus to determine the effect of various hormones and growth promoting factors by estimating the increase in cell number (20,32,46).…”

Section: Hormonal Requirements For Cell Proliferationmentioning

confidence: 99%

Cell proliferation and milk protein gene expression in rabbit mammary cell cultures

Ym¹,

Mt²,

Farinon³

et al. 1983

The Journal of Cell Biology

View full text Add to dashboard Cite

We analyzed the synthesis of DNA, the rate of cell proliferation, and the expression of milk protein genes in mammary cells grown as primary cultures on or in collagen gels in chemically defined media. We assessed DNA synthesis and cell growth, measured by [aH]-thymidine incorporation into acid-insoluble material, DNA content, and cell counts, in a progesterone-and prolactin-containing medium. In some experiments, cultures were pulsed for I h with [3H]thymidine and dissociated into individual cells which were cytocentrifuged and processed for immunocytochemistry and autoradiography. We analyzed expression of milk protein genes at the transcriptional, translational, and posttranslational levels in a progesteronedepleted medium in the presence or absence of prolactin. We measured protein secretion by radioimmunoassays with antisera directed against caseins, ~-lactalbumin and milk transferrin. We determined protein synthesis by incorporating radio-labeled amino acids into acid-precipitable material and by immunoprecipitating biosynthetically labeled milk proteins. We assessed the accumulation of casein mRNA by hybridizing total cellular RNA extracted from cultured cells with 32p-labeled casein cDNA probes. On attached collagen gels, the cells synthesized DNA and replicated until they became confluent. The overall protein synthetic activity was low, and no milk proteins were synthesized or secreted even in the presence of prolactin. The block in milk protein gene expression was not restricted to translational or posttranslational events but also included transcription, since no casein mRNA accumulated in these cells. On floating gels, protein synthesis was threefold higher than in cells from attached gels. Overall protein synthesis as well as casein and ~-lactalbumin synthesis and secretion were prolactindependent with maximal stimulation at around 10 -9 M. A marked inhibition occurred at higher hormone concentrations. Casein mRNA accumulated in these cells, provided prolactin was present in the medium. In contrast, these cells did not synthesize DNA, nor did they replicate. In embedding gels, the rate of cell proliferation was exponential over 25 d with a doubling time of ~70 h. The overall protein synthesis increase was parallel in time with the increase in cell number. Caseins and ~-lactalbumin (in contrast to transferrin) were synthesized only in the presence of prolactin. We observed the same hormone dependency as with cells growing on floating gels. The number of casein-and transferrin-positive cells was measured after dissociating the cell cultures. At day 12, 60% of the total cells stored transferrin in small cytoplasmic vesicles, whereas only 25% of the cells accumulated casein. Differences in the organization and in the shape of mammary cells depending on cell surface conditions suggest that the geometry of the cells, their interaction with extracellular matrix constitutents, and cellto-cell interactions play a role in the expression of two mammary functions: DNA synthesis and growth, as well as m...

show abstract

Section: Hormonal Requirements For Cell Proliferationmentioning

confidence: 99%

Cell proliferation and milk protein gene expression in rabbit mammary cell cultures

Ym¹,

Mt²,

Farinon³

et al. 1983

The Journal of Cell Biology

View full text Add to dashboard Cite

show abstract

“…Proliferation of epithelial cells can also be studied on attached collagen gels before confluence is attained (Mackenzie et al, 1982) and by embedding cells in collagen when branching duct-like structures form (Yang et al, 1980;Imagawa et al, 1982;McGrath, 1987). Other recently developed methods include the use of feeder layers of 3T3-L1 adipocytes (Levine & Stockdale, 1984) and extracellular matrix extracted from mammary tissue (Wicha et al, 1982).…”

Section: Culture In Vitromentioning

confidence: 99%

Growth factors in mammary gland function

Forsyth¹

1989

Reproduction

View full text Add to dashboard Cite

“…A whole mammary gland culture system has been established by Ichinose and Nandi (69); more recently, several methods for primary culture of mammary epithelial cells have been developed. Several nonplastic substrates, such as collagen gels (70)(71)(72)(73)(74)(75), extracellular matrix (76,77), and L3TL1 adipocytes (52,78) have been used for their capacity to support and promote mammary epithelial growth and differentiation. Cell culture offers the opportunity to distinguish the activity of the different cell populations in the mammary gland; whole gland cultures are particularly suitable in studying the structural changes occurring during the various phases of development.…”

Section: Biochemical Markers Of Differentiation In Vitromentioning

confidence: 99%

Growth Control and Differentiation in Mammary Epithelial Cells

Borellini¹,

Oka²

1989

Environmental Health Perspectives

View full text Add to dashboard Cite

Growth and differentiation of the mammary gland are controlled by various hormones and other environmental factors The role of hormones and growth factors in mammary development is discussed with regard to animal species, physiological stages, and the various experimental systems in vitro. In the female embryo, mammary morphogenesis is induced by the mesenchyme and is hormone independent, whereas androgens cause the partial necrosis of mammary epithelium in the male. Ductal growth during adolescence requires estrogen and prolactin or growth hormone. During pregnancy, progesterone participates in the development of the lobuloaveolar structure of the gland. After parturition, changes in the hormonal environment lead to production and secretion of milk. Proliferation and differentiation of mammary epithelium can be induced in culture systems. Insulin and epidermal growth factor (EGF) stimulate mammary cell proliferation in vitro. EGF is required for the optimal growth of the mammary gland during pregnancy. EGF also appears to play an important role in mammary tumorigenesis in certain mouse strains. Production of milk proteins can be induced in vitro by the synergistic interactions of prolactin, insulin, and glucocorticoids and is inhibited by EGF and progesterone. Complete or partial sequencing of several milk protein genes and comparative analysis have led to identification of a sequence of high homology and conservation in the 5 'flanking region that is likely to be involved in the regulation of milk protein gene expression.

show abstract

Serum-free growth of normal and tumor mouse mammary epithelial cells in primary culture.

Cited by 106 publications

References 28 publications

Cell proliferation and milk protein gene expression in rabbit mammary cell cultures

Cell proliferation and milk protein gene expression in rabbit mammary cell cultures

Growth factors in mammary gland function

Growth Control and Differentiation in Mammary Epithelial Cells

Contact Info

Product

Resources

About