Establishment and characterization of bovine mammary epithelial cell lines

Gibson, C A; Vega, Juan R.; Baumrucker, Craig R.; Oakley, Cori S.; Welsch, Clifford W.

doi:10.1007/bf02634376

Cited by 18 publications

(15 citation statements)

References 23 publications

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“…It contains principally laminin and collagen IV as well as a number of growth factors (McGuire & Seeds, 1989). Other workers have used floating collagen gels extracted from rat or calf tails (Talhouk et al 1990(Talhouk et al , 1993Gibson et al 1991 ;Huynh et al 1991). Using collagen gels, the best results in terms of differentiation have been achieved when the cells are allowed to form a confluent layer, and the collagen is then released from the bottom of the well causing the collagen to shrink, which in turn stimulates the cells to differentiate (Emerman & Pitelka, 1977).…”

Section: mentioning

confidence: 99%

“…Primary cell cultures have also been used with some success for short-term experiments ; with an appropriate hormonal milieu and extracellular matrix, primary MEC can be stimulated to re-group and re-differentiate into structures that resemble alveolar organoids and secrete milk proteins (Talhouk et al 1993). A number of immortal bovine MEC lines also exist (Schmid et al 1983 ;Gibson et al 1991 ;Huynh et al 1991 ;Huynh and Pollak, 1995 ;Zavizion et al 1996). However, in general, the level of milk protein synthesis of these transformed bovine cell lines is either very low or nonexistent, and if it does occur, it is independent of some or all of the in vivo factors (hormones or extracellular matrix) known to regulate lactogenesis (Huynh et al 1991).…”

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confidence: 99%

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In vitro differentiation of a cloned bovine mammary epithelial cell

Rose

Aso

Yonekura

et al. 2002

Journal of Dairy Research

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Rose, M. T., Aso, H., Yonekura, S., Komatsu, T., Hagino, A., Ozutsumi, K., Obara, Y. (2002). In vitro differentiation of a cloned bovine mammary epithelial cell.? Journal of Dairy Research, 69, (3), 345-355The aim of the study was to establish in vitro a bovine mammary epithelial cell (MEC) clone, able to respond to mitogenic growth factors and to lactogenic hormones. Mammary tissue from a 200-d pregnant Holstein cow was used as a source of MEC, from which a clone was established through a process of limiting dilution. When plated on plastic, the cells assumed a monolayer, cobblestone, epithelial-like morphology, with close contact between cells. Inclusion of IGF-1 and EGF in the media significantly increased the number of cells 5 d after plating. All cells stained strongly for cytokeratin and moderately for vimentin at young and old passage stages, indicating the epithelial nature of this cell clone. When the cells were plated at a high density on a thin layer of a commercial extracellular matrix preparation (Matrigel), lobular, alveoli-like structures developed within approximately 5 d, with a clearly visible lumen. When cells were plated onto Matrigel in differentiation media (containing lactogenic hormones), detectable quantities of ?-casein were present in the media and particularly on the lumen side of the structures. Omission of one of the lactogenic hormones (insulin, prolactin or hydrocortisone) reduced ?-casein release to the limit of detection of the assay used. Lactoferrin was also produced when the cells were plated on Matrigel, again principally on the lumen side of the lobules, though this was independent of the lactogenic hormones. By passage 40, the cells had senesced, and it was not possible to induce ?-casein or lactoferrin production. This study notes the establishment of a functional bovine mammary epithelial cell clone, which is responsive to mitogenic and lactogenic hormones and an extracellular matrix.Peer reviewe

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Section: mentioning

confidence: 99%

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confidence: 99%

In vitro differentiation of a cloned bovine mammary epithelial cell

Rose

Aso

Yonekura

et al. 2002

Journal of Dairy Research

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“…Several ruminant mammary epithelial cell lines (five bovine, two ovine and one caprine) have been established and well-characterized [2][3][4][5][6][7][8][9]. Bovine cell lines BMEC+H (Bovine Mammary Epithelial Cells of the Hormone-adapted) [4], HH2A (spontaneously immortalized bovine mammary epithelial cell line) [10], and ET-C (epithelial and myoepithelial-like characteristics) [11] do not express lactationspecific proteins.…”

Section: Introductionmentioning

confidence: 99%

Transcriptomics Comparisons of Mac-T cells Versus Mammary Tissue during Late Pregnancy and Peak Lactation

Hosseini¹,

Sharma²,

Bionaz³

et al. 2013

J Adv Dairy Res

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Epithelial cell cultures, including immortalized cells, have been used extensively to help answer basic questions of mammary gland physiology under normal or diseased conditions. The usefulness of using cell culture depends on the degree of identify compared to the in vivo mammary epithelium. Our objective was to compare the transcriptome by means of microarray and qPCR of immortalized bovine mammary epithelial cells (Mac-T) cultivated in plastic (i.e., 2 dimensional system) vs. mammary tissue obtained during late-pregnancy (-30 DIM) and peak lactation (+60 DIM). Functional analysis of microarray data was performed using enrichment analysis tools and the Dynamic Impact Approach (DIA). Overall, between 37% (vs. +60 DIM) and 44% (vs.-30 DIM) of the measured annotated genes were deemed as differentially expressed (DEG) between Mac-T cells and mammary tissue. These data, together with the bioinformatics results of the genes non-differentially expressed, indicated a larger overall similarity between Mac-T cells and lactating than non-lactating mammary tissue. However, Mac-T cells had a lower expression measured by qPCR of genes involved in milk synthesis compared to mammary tissue. The functional analysis of DEG further supported the poor lactation phenotype of Mac-T cells compared with mammary tissue. The bioinformatics analyses of DEG suggested that Mac-T cells cultivated on plastic had greater reliance on glucose, amino acids, and fatty acids for production of energy, greater cell-to-cell interactions, and more prone to react to inflammatory mediators relative to mammary tissue. Despites these differences, data suggest that Mac-T cells might be adequate for studying milk protein regulation. For studies of milk fat regulation Mac-T cells might be not as sensitive as the mammary tissue, particularly for its lower expression of PPARG and lower induction of PPARγ-and LXR-related pathways. Overall, this study revealed a marked difference in the transcriptome signatures between mammary tissue and Mac-T cells cultivated on plastic.

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“…The mammary gland cell shows cycles of mammary gland development, differentiation, and apoptosis during the span of its physiologic cycle and during pregnancy. Unfortunately, mammary gland cells have a limited lifespan in culture: approximately 10–15 passages (Gibson et al. 1991).…”

Section: Introductionmentioning

confidence: 99%

Recombinant adenovirus‐mediated human telomerase reverse transcriptase gene can stimulate cell proliferation and maintain primitive characteristics in bovine mammary gland epithelial cells

Zheng

Lan

et al. 2011

Dev Growth Differ

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The human telomerase reverse transcriptase (hTERT) gene has been used to stimulate the proliferation of most types of human cells. The present study was designed to evaluate the feasibility and efficiency of adenovirusmediated hTERT in the proliferation of bovine mammary gland epithelial cells (bMGEs). A plasmid and an adenovirus vector that carried hTERT, namely pEGFP-hTERT and Ad-hTERT, were constructed and transfected into bMGEs, respectively. In order to select the best strategy for stimulating cell proliferation, the adenovirusand plasmid-mediated hTERT were compared in terms of the positive cloning and transgenic efficiency. The results showed that only Ad-hTERT had high infection efficiency and produced a positive polyclone population (hTERT-bMGEs). The characteristics of the hTERT-bMGEs were investigated with further analysis by reverse transcription-polymerase chain reaction (RT-PCR), western blotting, proliferation assays, and flow cytometry, which showed that hTERT facilitated strong cell proliferation. Real-time quantitative PCR showed a normal level of expression of beta-casein, the caspase-8 and c-myc proto-oncogene, and immunofluorescence demonstrated the properties of the epithelial cells. In conclusion, the adenovirus-mediated hTERT gene could not only extend the cell lifespan, but also maintained the primary characteristics of the cells. It may be possible to extend the use of a wide variety of non-human mammalian cells in this way. This study has provided additional insight into the mechanism of cell proliferation by demonstrating the lack of integration of the adenovirus-mediated hTERT gene into the mammalian genome.

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Establishment and characterization of bovine mammary epithelial cell lines

Cited by 18 publications

References 23 publications

In vitro differentiation of a cloned bovine mammary epithelial cell

In vitro differentiation of a cloned bovine mammary epithelial cell

Transcriptomics Comparisons of Mac-T cells Versus Mammary Tissue during Late Pregnancy and Peak Lactation

Recombinant adenovirus‐mediated human telomerase reverse transcriptase gene can stimulate cell proliferation and maintain primitive characteristics in bovine mammary gland epithelial cells

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