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V Anand N Dogra S Singh SN Kumar MK Jena D Malakar AK Dang BP Mishra TK Mukhopadhyay JK Kaushik AK Mohanty 《PloS one》2012,7(7):e40469
Background
The objective of this study was to establish the buffalo mammary epithelial cell line (BuMEC) and characterize its mammary specific functions.Methodology
Buffalo mammary tissue collected from the slaughter house was processed enzymatically to obtain a heterogenous population of cells containing both epithelial and fibroblasts cells. Epithelial cells were purified by selective trypsinization and were grown in a plastic substratum. The purified mammary epithelial cells (MECs) after several passages were characterized for mammary specific functions by immunocytochemistry, RT-PCR and western blot.Principal Findings
The established buffalo mammary epithelial cell line (BuMEC) exhibited epithelial cell characteristics by immunostaining positively with cytokeratin 18 and negatively with vimentin. The BuMEC maintained the characteristics of its functional differentiation by expression of β-casein, κ-casein, butyrophilin and lactoferrin. BuMEC had normal growth properties and maintained diploid chromosome number (2n = 50) before and after cryopreservation. A spontaneously immortalized buffalo mammary epithelial cell line was established after 20 passages and was continuously subcultured for more than 60 passages without senescence.Conclusions
We have established a buffalo mammary epithelial cell line that can be used as a model system for studying mammary gland functions. 相似文献3.
本研究旨在建立牦牛乳腺上皮细胞体外培养体系。采用胶原酶消化法成功地建立了牦牛乳腺上皮细胞系(YMEC),通过免疫细胞化学、超微结构观察和RT-PCR 法对YMEC 细胞进行了鉴定,并研究了其形态、活力、生长曲线以及核型等生物学特性。结果表明,YMEC 细胞染色体2n = 60,群体倍增时间为45 ~ 48 h,持续培养25 代后出现细胞分化;细胞呈典型的“铺路石样”形态,其表面有丰富的微绒毛,细胞质内含丰富的线粒体和粗面内质网。污染检测结果为阴性。在激素诱导培养时,检测到了β - 酪蛋白mRNA 的表达。表明本研究成功建立了保留泌乳功能的牦牛乳腺上皮细胞系,为研究牦牛乳腺上皮细胞的功能提供了理想的工具。 相似文献
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C. Anne Gibson Juan R. Vega Craig R. Baumrucker Cori S. Oakley Clifford W. Welsch 《In vitro cellular & developmental biology. Animal》1991,27(7):585-594
Summary One bovine mammary epithelial cell clone, designated PS-BME-C1, and two bovine mammary epithelial cell lines, designated PS-BME-L6
and PS-BME-L7, were derived from mammary tissue of a pregnant (270 day) Holstein cow. The cells exhibit the distinctive morphologic
characteristics of mammary epithelial cells and express the milk fat globule membrane protein, PAS-III. They form domes when
cultured on plastic substrata and acinilike aggregates when cultured on a collagen matrix. These cells are capable of synthesizing
and secretingα-lactalbumin andα-s1-casein when cultured on a collagen matrix in the presence of insulin, cortisol, and prolactin. The cells have a near-normal
diploid number and do not grow in suspension culture. When transplanted to the cleared mammary fat pads of female athymic
nude mice, the cells readily proliferate forming noninvasive palpable spherical cellular masses within 8 wk after inoculation.
The cells may become a useful tool to study the regulation of ruminant mammary epithelial cell growth and differentation.
This work was supported by the Pennsylvania State University Experiment Station.
The PS-BME cells are the property of The Pennsylvania Research Corporation. Scientists interested in obtaining the PS-BME
clone or cell lines for their research may request them from the corresponding author. 相似文献
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Establishment and characterization of a bovine mammary epithelial cell line with unique properties 总被引:8,自引:0,他引:8
Boris Zavizion Marilyn van Duffelen Warren Schaeffer Ioannis Politis 《In vitro cellular & developmental biology. Animal》1996,32(3):138-148
Summary Clonal cell lines (BME-UV) were established from primary epithelial cells by stable transfection with a plasmid, carrying
the sequence of the simian virus 40 early region mutant tsA58, encoding the thermolabile large T antigen. The BME-UV cells
have undergone more than 300 population doublings and produce intranuclear large T antigen. At low confluency, growing islands
of cells are apparent exhibiting the characteristic cobblestone morphology of epithelial cells. The BME-UV cells expressed
functional markers such as microvilli and desmosomes and biochemical markers of mammary epithelial cells such as a repertoire
of cytokeratins. The BME-UV cells are capable of synthesizing low levels of α-lactalbumin and α8l (50 ng/ml of medium/24 h). One of the cell lines, BME-UV1 showed enhanced proliferation in the presence of epidermal growth
factor (EGF) and insulinlike growth factor I (IGF-I). The BME-UV1 cell line is the only known bovine mammary epithelial cell
line responsive to EGF. The BME-UV cells grown on collagen at low confluency are capable of developing very long projections
that most likely allow for communication between cells at a distance from each other. The BME-UV cells may become a valid
model system to examine bovine mammary epithelial proliferation and differentiation and cell-to-cell communication. 相似文献
7.
Establishment and characterization of a lactating dairy goat mammary gland epithelial cell line 总被引:1,自引:0,他引:1
To study milk synthesis in dairy goat mammary gland, we had established an in vitro lactating dairy goat mammary epithelial
cell (DGMEC) line. Mammary tissues of Guan Zhong dairy goats at 35 d of lactation were dispersed and cultured in a medium
containing epithelial growth factor, insulin-like growth factor-1, insulin transferrin serum, and fetal bovine serum. Epithelial
cells were enriched by digesting with 0.25% trypsin repeatedly to remove fibroblast cells and were identified as epithelial
origin by staining with antibody against cytokeratine 18. The DGMECs displayed monolayer, cobble-stone, epithelial-like morphology,
and formed alveoli-like structures and island monolayer aggregates which were the typical characteristics of mammary epithelial
cells. A one-half logarithmically growth curve and cytoplasmic lipid droplets in these cells were observed. In this paper,
we also studied the lactating function of DGMECs. Results showed that DGMECs could secrete lactose and β-casein. Lactating
function of the cells had no obvious change after 48 h treated by insulin, while prolactin could obviously raise the secretion
of milk proteins and lactose. 相似文献
8.
Establishment and initial characterization of the ovine mammary epithelial cell line nish 总被引:4,自引:0,他引:4
Neta Ilan Itamar Barash Elisha Gootwine Moshe Shani 《In vitro cellular & developmental biology. Animal》1998,34(4):326-332
Summary Analysis of the molecular mechanisms involved in the differentiation and formation of the characteristic three-dimensional
structures of the developing mammary gland of the major milk-producing livestock (ducts, end buds, and alveoli) requires in vitro model cell cultures. The few cell lines that have been established from dairy animals do not fully reproduce the entire program
of mammary differentiation. Here we present the initial characterization of a unique mammary epithelial cell line derived
spontaneously from midpregnant sheep (NISH). These cells form in vitro functional structures resembling ducts, lateral buds, and alveoli that secrete β-lactoglobulin (BLG) in an ECM (extracellular
matrix)-dependent manner. Interestingly, the presence of growth hormone dramatically increased BLG secretion from NISH cells
cultured on ECM. It appears that GH is required not only to establish the structural organization but also is continuously
needed to maintain BLG expression. Stable transfection of NISH cells with BLG/Human Serum Albumin (HSA) hybrid gene constructs
revealed that the relative level of expression was comparable to the in vivo secretion of HSA in transgenic mice carrying these gene sequences. No expression could be detected in cells transfected with
hybrid genes carrying either HSA cDNA or the entire HSA gene, and HSA expression was dependent on the presence of intronic
sequences. These results demonstrate that NISH cells may prove a useful tool for studying the differentiation and organogenesis
of mammary epithelial cells under defined culture conditions. Furthermore, transfected NISH cells may be an alternative for
the transgenic mouse model in evaluating the potential of gene constructs to be efficiently expressed in the mammary gland
of transgenic farm animals. 相似文献
9.
Boris Zavizion Marilyn van Duffelen Warren Schaeffer Ioannis Politis 《In vitro cellular & developmental biology. Animal》1996,32(3):149-158
Summary The thermolabile large T-antigen, encoded by the simian virus 40 early region mutant tsA58, was used to establish clonal cell
lines (BMM-UV) from primary bovine myoepithelial cells. The BMM-UV cells have undergone more than 300 population doublings
without any signs of senescence, and they contain the intranuclear large T antigen. At low confluency, they grow in a spindlelike
manner and develop very long projections that most likely allow for communication of cells at a distance from each other.
Establishment results in a decrease in the number of cells that contract in response to oxytocin compared with the parental
nontransfected cells (20% versus 45%). Oxytocin responsiveness of BMM-UV cells increases when the cells are cultured in a
medium supplemented with staphylococcal proteases. Proliferation of BMM-UV cells increases when they are cultured in the presence
of epidermal growth factor (10 ng/ml) or insulinlike growth factor I (50 ng/ml). The BMM-UV cells may become a useful model
to study growth properties, cell-to-cell communication, and the function of bovine mammary myoepithelial cells. 相似文献
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Yoji Nagashima Yoshiharu Ohaki Makoto Umeda Mitsuo Oshimura Kazuaki Misugi 《Virchows Archiv. B, Cell pathology including molecular pathology》1989,57(1):383-392
A new cell line designated ENU-T-1 has been established from a xenotransplanted experimental rat nephroblastoma. The cultured
cells are spindle-shaped or polygonal and are arranged in a wavy fashion morphologically similar to cultured embryonal renal
epithelial cells. The cells exhibit a number of epithelial characteristics. Enzyme histochemistry gives positive reactions
for gammaglutamyltranspeptidase and alkaline phosphatase, both of which are present in renal tubular epithelial cells. Immunofluorescence
studies show positive reactions for vimentin and cytokeratin. When inoculated into athymic nude mice, the cultured cells form
tumors composed of sheets of epithelial cells with focal tubular formation.
This cell line may be of value in studying differentiation of nephroblastoma, and possibly normal nephrogenesis. 相似文献
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Y Nagashima Y Ohaki M Umeda M Oshimura K Misugi 《Virchows Archiv. B, Cell pathology including molecular pathology》1989,57(6):383-392
A new cell line designated ENU-T-1 has been established from a xenotransplanted experimental rat nephroblastoma. The cultured cells are spindle-shaped or polygonal and are arranged in a wavy fashion morphologically similar to cultured embryonal renal epithelial cells. The cells exhibit a number of epithelial characteristics. Enzyme histochemistry gives positive reactions for gamma-glutamyltranspeptidase and alkaline phosphatase, both of which are present in renal tubular epithelial cells. Immunofluorescence studies show positive reactions for vimentin and cytokeratin. When inoculated into athymic nude mice, the cultured cells form tumors composed of sheets of epithelial cells with focal tubular formation. This cell line may be of value in studying differentiation of nephroblastoma, and possibly normal nephrogenesis. 相似文献
16.
Establishment and characterization of a human chorionic gonadotropin (hCG) producing cell line (RTSG) from an ovarian epithelial cancer 总被引:3,自引:0,他引:3
Y Udagawa S Nozawa K Chin M Sakayori M Mikami K Ohta K Tsukazaki K Kiguchi R Iizuka 《Human cell》1990,3(1):70-75
A new cell line designated RTSG established in vitro from the pleural effusion of a patient with metastatic ovarian epithelial cancer has been subcultured 46 times for more than 2 years. The cells grew in a monolayered sheet, showing a tendency to pile up, with the population doubling in 48 hrs. Electron-microscopically, desmosomes were characteristically observed, suggesting the cells were of epithelial origin. Chromosomal analysis revealed aneuploidy with a tetraploid mode. The heterotransplanted tumors in nude mice were histopathologically classified as a poorly differentiated adenocarcinoma, whereas the original tumor consisted mainly of mucinous and serous cystadenocarcinoma and only partly of poorly differentiated adenocarcinoma. The cells secreted hCG (38.8 mIU/day/10(6) cells) and beta-hCG (6.1 ng/day/10(6) cells) in spent medium. Immunocytologic +-and-histochemical staining for tumor markers of the original tumor, the cultured cells and the transplanted tumors also revealed the localization of not only hCG and beta-hCG but also CA19-9 and CA-125 whose values had been elevated in the preoperative serum (hCG: 10 mIU/ml, CA19-9: 6,400 U/ml, CA-125: 225 U/ml). Results of PAS, Alcian-blue and Mucicarmine strains indicated that most of the PAS-positive substances in the cultured cells and the transplanted tumors were consistent with glycogen while the original tumor mainly contained mucin except for the lesion of poorly differentiated adenocarcinoma with glycogen. These results suggested that the cultured cells might originate from poorly differentiated adenocarcinoma cells in the original tumor. 相似文献
17.
Human gestational choriocarcinoma cell line (HOCC) was established from the mouse graft of choriocarcinoma. The HOCC cells were spindle or polygonal in shape and multi-nucleated giant cells, showing neoplastic and pleomorphic features. The cell proliferated stably, and the population doubling time was about 32 hours. The chromosome numbers showed a wide distribution of aneuploidy, the mode was in hypertriploid range and the marker chromosomes were recognized in the several generations. Heterotransplantation was easy, and subcutaneous transplantation of 1 X 10(7) cells in nude mouse formed a tumor composed of choriocarcinoma. It is most noteworthy characteristic of the cell line that it produced human chorionic gonadotropin (hCG) in an in vitro culture system and in vivo in nude mice. 相似文献
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Isamu Ishiwata Chieko Ishiwata Akira Tsuiki Kazushige Kiguchi Hisashi Hashimoto Hiroshi Ishikawa 《Human cell》2008,17(1):33-41
Human gestational choriocarcinoma cell line (HOCC) was established from the mouse graft of choriocarcinorna. The HOCC cells were spindle or polygonal in shape and multi-nucleated giant cells, showing neoplastic and pleomorphic features. The cell proliferated stably, and the population doubling time was about 32 hours. The chromosome numbers showed a wide distribution of aneuploidy, the mode was in hypertriploid range and the marker chromosomes were recognized in the several generations. Heterotransplantation was easy, and subcutaneous transplantation of 1 × 107 cells in nude mouse formed a tumor composed of choriocarcinoma. It is most noteworthy characteristic of the cell line that it produced human chorionic gonadotropin (hCG) in an in vitro culture system and in vivo in nude mice. 相似文献
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Ishiwata I Ishiwata C Anzo M Minami R Kiguchi K Tachibana T Ishikawa H 《Human cell》2003,16(4):231-239
A cell line designated "HMMME" was established from the pleural fluids of a malignant mesothelioma patient. This line grew well without interruption for 12 years and was subcultured over 200 times. The cells were spindle and roundish in shape and displayed a monolayer sheet in an epithelial pavement cell arrangement. They were neoplastic, had pleomorphic features, and easily formed multilayering without contact inhibition. The cell cytoplasm was strongly positive against anti-vimentin, anti-calretinin and anti-pan-keratin, but negative against anti-BerEP4. The cells proliferated rapidly, and the population doubling time was about 42 hours. Their chromosome number showed a wide distribution of aneuploidy with a mode in the diploid range; many marker chromosomes were observed. The cultured cells were easily transplanted into the subcutaneous of nude mice and produced a tumor classified as a malignant mesothelioma. 相似文献
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I Ono I Ishiwata T Nakaguchi M Soma N Tokita C Ishiwata M Mukai S Nozawa H Ishikawa 《Human cell》1989,2(3):272-277
The malignant schwannoma cell line (HKMS) was established from the subcutaneous tumor of Axilla region of a 48-year-old Japanese woman. The HKMS line has the following biological properties. 1. The HKMS cells were spindle in shape and showed neoplastic and pleomorphic features. The monolayer sheet of HKMS cells showed the resemble cell-arrangement with that of the original tumor tissue. 2. The cells showed a stable growth and the serial passages were successively carried out 150 times within 3 years. Their population doubling time is about 40 hours. 3. The chromosome number varied widely, and the modal number was stable at the 78-80. The marker chromosomes were present. 4. The cells were transplanted into the subcutis of nude mice and produced the malignant schwannoma. 相似文献