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1.
Antisera produced to mammary adipose cells from midpregnant BALC/c females can be used to distinguish mammary adipose cells from mammary epithelial cells and fibroblasts. The mammary adipose membrane antigen detected by indirect immunofluorescence was found in adipose cells from (a) mammary glands of virgin, midpregnant and lactating mice; (b) mammary fat pads that had been surgically cleared of glandular elements; and (c) epididymis. In all tissues, this cell-surface antigen was removed by the enzymatic action used to dissociate the cells from the tissues and was shown to be fully restored when cells were cultured for 48 hr. 相似文献
2.
Quantitation of human mammary epithelial antigens in cells cultured from normal and cancerous breast tissues 总被引:1,自引:0,他引:1
Masao Sasaki J. A. Peterson R. L. Ceriani 《In vitro cellular & developmental biology. Plant》1981,17(2):150-158
Summary A sensitive radioimmunoassay technique was developed to quantitatite the level of human breast celltype specific antigens
on cells from normal breast and from various established cell lines of breast and nonbreast origins. Polyacrylamide gel electrophoresis
revealed four major proteinaceous components (150,000; 75,000; 60,000; and 48,000) in human milk fat globule membranes that
were used to immunize rabbits in order to elicit antimammary epithelial cell antibody. Antisera obtained were rendered specific
by abosorptions and were able to recognize three specific mammary epithelial components of the breast epithelial cell. Human
mammary epithelial (HME) antigen expression was highest (1290 ng/106 cells) in normal breast epithelial cells from primary cultures of normal breasts. Lower levels (range: 955 to 330 ng/106 cells) were found in breast epithelial cells from cell lines established from cancerous breast tissue. Cells of nonbreast
origins as well as fibroblasts from breast gave much lower values (less than 30 ng/106 cells). On treatment, with trypsin, of two breast epithelial cell lines (MDA-MB-157 and MCF-7) 80 to 85% of their HME antigen
expression was lost, suggesting that a majority of these breast antigens reside on the cell surface.
This work was Supported by Grant PTD-99 from the American Cancer Society, Grant CA19455 and CA20286 from the National Cancer
Institute, and Biomedical Research Support Grant RR05467 from the National Institutes of Health. Most cells used in the present
study were produced with support from National Cancer Institute Contract Y01-CP8-0500, Biological Carcinogenesis Branch, Division
of Cancer Cause and Prevention, under the auspices of the Office of Naval Research and the Regents of the University of California. 相似文献
3.
Electrical coupling has been observed between cultured cells of the mouse mammary gland in five distinct physiological or pathological states. We have employed young primary cultures of cells dissociated from the following tissues: normal glands from young virgin or midpregnant females, hyperplastic alveolar nodules (believed to be precancerous) transplanted in gland-free mammary fat pads, and spontaneous mammary adenocarcinomas and their pulmonary metastases. All successfully impaled pairs of cells (a total of 97 pairs) were found to be ionically coupled. Furthermore, in normal and tumor cell cultures, electrical coupling was observed between dome-dome and dome-nondome cell pairs. This study correlates with electronmicroscopic studies of fresh normal, hyperplastic, and tumor samples, which show the presence of gap junctions in all three. 相似文献
4.
Sheldon S. Shen Susan T. Hamamoto Dorothy R. Pitelka 《The Journal of membrane biology》1976,29(1):373-382
Summary Electrical coupling has been observed between cultured cells of the mouse mammary gland in five distinct physiological or pathological states. We have employed young primary cultures of cells dissociated from the following tissues: normal glands from young virgin or midpregnant females, hyperplastic alveolar nodules (believed, to be precancerous) transplanted in gland-free mammary fat pads, and spontaneous mammary adenocarcinomas and their pulmonary metastases. All successfully impaled pairs of cells (a total of 97 pairs) were found to be ionically coupled. Furthermore, in normal and tumor cell cultures, electrical coupling was observed between dome-dome and dome-nondome cell pairs. This study correlates with electronmicroscopic studies of fresh normal, hyperplastic, and tumor samples, which show the presence of gap junctions in all three. 相似文献
5.
Leonard A. Cohen Julian Tsuang Po C. Chan 《In vitro cellular & developmental biology. Plant》1974,10(1-2):51-62
Summary The characteristics of normal mammary epithelial and 7,12-dimethylbenz[a]anthracene (DMBA)-induced adenocarcinoma cells derived
from rats and grown in monolayer culture were compared. Normal mammary epithelial cells exhibited different morphology and
agglutinability by plant lectins, slower growth rate, and lower saturation density and cloning efficiency. In addition, the
normal cells were sensitive to the toxic effect of DMBA, and were unable to grow in soft agar or to form tumors, when inoculated
into newborn Sparague-Dawley rats. The converse was true in each case for the adenocarcinoma cells.
Supported by Public Health Service Research Grant CA 01237603 from the National Cancer Institute
Portions of this paper were presented at the 65th Annual Meeting of the American Association for Cancer Research at Houston,
Texas, 1974. 相似文献
6.
Elevated levels of xanthine oxidase were found in (1) lactating mouse mammary glands, compared with virgin and midpregnant glands; and (2) primary mouse mammary cells cultured on floating collagen gels, compared with non-secretory cells on attached gels. In primary culture, increase in xanthine oxidase activity above a basal level coincided with secretory activity as measured by casein production; intracellular levels of casein and xanthine oxidase showed a high degree of correspondence. It is suggested that xanthine oxidase levels can be used as an indicator of in vivo and in vitro secretory differentiation in mammary epithelial cells. 相似文献
7.
Rillema JA Hill MA 《American journal of physiology. Endocrinology and metabolism》2003,284(1):E25-E28
Iodide is an essential constituent of milk that is present in concentrations more than an order of magnitude higher than in the maternal plasma. Earlier, a sodium-iodide symporter was identified in the mammary gland; this transporter is presumed to take iodide from the maternal plasma into the alveolar epithelial cells of the mammary gland. We now report the existence of a second iodide transporter, pendrin, which is also essential for iodide accumulation in milk. Via Western blotting methods, high levels of the transporter were detected in lactating tissues; lesser amounts were found in tissues from midpregnant and virgin mice. Prolactin, at physiological concentrations, stimulated the expression of the pendrin transporter in cultured mammary tissues taken from 12- to 14-day-pregnant mice. The prolactin effect on iodide uptake into cultured mammary tissues was abolished by pendrin transport inhibitors, including DIDS, furosemide, and probenecid. These studies suggest that the prolactin stimulation of pendrin activity is an essential element in the prolactin stimulation of iodide uptake into milk. 相似文献
8.
Growth inhibitors in plasma derived human serum 总被引:1,自引:0,他引:1
Dennis A. Pigott Margaret A. Grimaldi Marie L. Dell’Aquila Edwin V. Gaffney 《In vitro cellular & developmental biology. Plant》1982,18(7):617-625
Summary It was reported previously that plasma derived human serum (PDS) inhibited the growth of cells established from malignant
human breast tissues and the MCF-7 cell line but did not inhibit the growth of cells from nonmalignant mammary tissues, including
the HBL-100 cell line. Plasma derived human serum was fractionated in the current study by molecular sieve chromatography
on Sephadex G-100 in an effort to characterize the factor(s) responsible for inhibition. Plasma derived human serum contained
several growth inhibitory fractions, which were designated G-1, G-2, G-3, and G-4. The G-1 was associated with the lipoproteins
and immunoglobulins of serum. The lipid portion of G-1 inhibited the growth of both MCF-7 and HBL-100 cells, whereas the protein
fraction contained a low activity factor directed against MCF-7 cells only. The G-2 also inhibited MCF-7 cell growth at a
low specific activity and was separated in the serum albumin fraction. The MCF-7 inhibitory activity in the G-3 fractions
from individual donors fluctuated with the level of activity in the starting sera. The cell specific G-3 components were purified
further by Sephadex G-100 superfine chromatography and gel electrophoresis. A tentative molecular weight of 50,000 was assigned
to the G-3 inhibitor. The G-4 fraction consisted of small molecular weight materials migrating in advance of the column volume,
which inhibited the growth of both cell lines.
This investigation was supported by Grant PDT-140 from the American Cancer Society, Inc., and PHS Grant CA30284 awarded by
the National Cancer Institute, Bethesda, Maryland. 相似文献
9.
In vitro analysis of proliferating epithelial cell populations from the mouse mammary gland: Fibroblast-free growth and serial passage 总被引:3,自引:0,他引:3
Michael T. White A. S. L. Hu Susan T. Hamamoto S. Nandi 《In vitro cellular & developmental biology. Plant》1978,14(3):271-281
Summary Normal and neoplastic mouse mammary epithelial cells were cultured in nutrient medium containing D-valine substituted for
L-valine. Fibroblast overgrowth was prevented and epithelial cell functions and morphology were retained in cultures maintained
in, D-valine medium up to 2 months. A nonenzymatic technique was devised to dissociate epithelial cell monolayers. The combined
use of this dissociation buffer and D-valine nutrient medium made it possible to passage serially normal and neoplastic mammary
epithelial cells. Normal cells were derived from mammary glands of animals stimulated with exogenous hormones for various
periods. The period of in vivo hormonal stimulation influenced the ability of normal mammary epithelial cells to attach and
proliferate in primary and serially passaged cultures. A greater proportion of cells derived from glands following 2 to 4
weeks of hormonal stimulation were recovered after replating and showed higher labeling indices during serial passage than
cells from unstimulated or 5- to 7-week stimulated groups.
This investigation was supported by Grant No. CA 05388 from the National Cancer Institute and by Cancer Research Funds of
the University of California. 相似文献
10.
Growth of normal human mammary cells in culture 总被引:27,自引:0,他引:27
M. Stampfer R. C. Hallowes A. J. Hackett 《In vitro cellular & developmental biology. Plant》1980,16(5):415-425
Summary Reduction mammoplasty tissue was used to obtain short-term cultures of human epithelial cell populations. Digestion of tissue
with collagenase and hyaluronidase resulted in cell clusters (organoids) resembling ductal and alveolar structures; these
could be separated by filtration from the stromal components. Epithelial outgrowth from these organoids was greatly enhanced
by the addition of conditioned medium from other human epithelial and myoepithelial cell lines. Additionally, the mammary
epithelial growth was stimulated by insulin, hydrocortisone, epidermal growth factor, and steroid hormones. With this enriched
nutritional environment, active cell division could be maintained for 1 to 3 months and cells could be serially subcultured
1 to 4 times.
This research was supported by Grant PDT-72 from the American Cancer Society and Grant CP-70510 from the National Institutes
of Health. 相似文献
11.
12.
13.
Summary Effects of three adrenal corticoids on in vitro mammary differentiation were compared in neonatally estrogenized (E) and uninjected
control (N) BALB/c Crgl female mice. E mice were injected with 10 μg of 17β-estradiol on each of the first 5 days of life.
At 4 weeks of age, all mice were pretreated with 1 μg 17β-estradiol and 1 mg progesterone for 9 consecutive days. Groups of
10 or more entire mammary glands then were organ-cultured for 5 days at 37°C on chemically defined medium in 95% O2/5% CO2 with various combinations of hormones: mammotropin (M), somatotropin (S), insulin (I), thyroxine (T); and corticosterone
(B), aldosterone (A), or cortisol (F). Differentiation followed a quantitative pattern of MSIT<BMSIT<AMSIT<FMSIT for both
E and N tissues. E tissues formed more alveoli and more lobules in vitro than N tissues with each of the corticoids tested.
These findings may have pathologic significance.
This work was supported in part by Public Health Service Grant CA-18285 from the National Cancer Institute and by Public Health
Service General Research Support Grant NIH-5-S01-RRO 5344-11. 相似文献
14.
Ceriani R. L. Peterson J. A. Abraham S. 《In vitro cellular & developmental biology. Plant》1978,14(11):887-894
Summary Treatment of mouse mammary epithelial cells (MMEC) with various enzymes used for dispersing and transferring cells results
in extensive digestion of materials on the cell surfaces. MMEC biosynthetially labeled with [3H]fucose, [14C]fucose and [3H]amino acids or with125I by the lactoperoxidase method were exposed to either collagenase plus hyaluronidase, followed by pronase, or to trypsin
in concentrations and conditions currently used for cell dispersion. Whereas the latter enzyme preparation solubilized 76%
of the trichloroacetic acid precipitable radioactive fucose and 96% of the protein-bound125I, collagenase plus hyaluronidase treatment released lesser amounts of each label. Subsequent treatment of the cells with
pronase removed additional surface-labeled materials, but the total amounts released were still less than when the trypsin
preparation alone was employed. Released cell surface materials were analyzed by gel chromatography. Some of the peaks obtained
also were examined by polyacrylamide gel electrophoresis. The labeled materials that remained attached to the MMEC after enzymatic
treatment were investigated by these two methods as well. We could show that collagenase plus hyaluronidase solubilized three
main glycoprotein components from the cell surface. In addition, we could show that the extensive cell surface damage caused
by these two enzyme preparations was due to the high proteolytic activity present in these preparations as judged by their
ability to hydrolyze rabbit gamma globulin labeled with125I. Even though their membranes were extensively damaged by the enzyme treatments, the dispersed cells could be cultured successfully
in vitro and could incorporated fucose into their surfaces in a manner similar to that by intact tissue. Through the use of
gel-filtration (cochromatography of [14C]fucose and [3H]fucose cell surface materials), we could demonstrate the identity of cell surface glycoproteins synthesized by cultured
cells and by intact tissue.
This work was supported by Grant Nos. CA 11736 and CA 19455 from the National Cancer Institute, and Biomedical Research Support
Grant No. RR05467 from the National Institutes of Health, DHEW. 相似文献
15.
Long-term organ culture of mouse mammary gland 总被引:1,自引:0,他引:1
J. W. Harbell P. D. Bowman J. M. Shannon C. W. Daniel 《In vitro cellular & developmental biology. Plant》1977,13(8):490-496
Summary A method for maintaining mouse mammary gland in organ culture for periods of at least 30 days is described. Strips of the
number four mammary glands were cultured in individual tubes while fully submerged in Medium 199 supplemented with insulin,
aldosterone, ovine prolactin and bovine growth hormone. Exchange processes were aided by slowly rotating the tubes during
culture. Mammary tissue from midpregnant BALB/c and virgin GR/A mice was induced to undergo lobulo-alveolar development, secrete
and remain differentiated and metabolically active for the period of culture. Cells of both the ductal and alveolar epithelium
continued to synthesize DNA and divide. The submerged roller-tube culture allows the use of larger pieces of tissue than can
be accommodated in static culture, and the technique may prove applicable to the culture of a variety of tissues. 相似文献
16.
D. Gaillard R. Négrel G. Serrero-Davé C. Cermolacce G. Ailhaud 《In vitro cellular & developmental biology. Plant》1984,20(2):79-88
Summary Ob17 is a clonal cell line isolated from the epididymal fat pad of C57 BL/6J ob/ob mouse that differentiates into adiposelike
cells in serum-supplemented medium. In serum-free medium, this cell line shows increased growth under the addition of insulin,
transferrin, fibroblast growth factor (FGF), and a factor present in extract of rat submaxillary gland (SMGE). This medium
is referred to as 4F. Epidermal growth factor or nerve growth factor cannot replace SMGE, whereas partially purified platelet
extract can substitute for FGF but only partially for SMGE. 4F Medium is able to support the proliferation of cells from other
established preadipocyte clonal lines, HGFu and 3T3-F442A, and also of preadipocyte cells isolated from the stromal-vascular
fraction of rat and mouse adipose tissues. In each case 4F medium is insufficient to support the differentiation of these
cells into adipocytes. Ob17 cells grown and maintained in serum-free hormone-supplemented medium retain the ability to convert
to adiposelike cells after serum addition. This serum requirement for differentiation cannot be substituted by the addition
of growth hormone or of other putative adipogenic factors, or both. The results are discussed with respect to the requirements
for growth and differentiation of the 3T3-L1 and 1246 preadipocyte cell lines previously described.
This work was supported by the “Centre National de la Recherche Scientifique” (Grant 1208-Biochimie du Développement and Grant
4162-Endocrinologie), by the “Ministère de la Recherce et de la Technologie” (Grant 81-L-1322), by the “Fondation pour la
Recherche Médicale,” by NATO (Grant 1704), and by the “Institut National de la Santé et de la Recherche Médicale” (Grant 827006). 相似文献
17.
E. M. Rivera S. D. Hill M. Taylor 《In vitro cellular & developmental biology. Plant》1981,17(2):159-166
Summary The purpose of this study was to determine whether culturing hyperplastic mammary nodules in hormone-free medium would enhance
their oncogenicity following subsequent transplantation into mammary fat pads. The underlying hypothesis is that the proliferation
of transformed cells within the nodules is inhibited by hormone-dependent normal cells also present in the nodules. Accordingly,
both primary hyperplastic nodules and tissues from a hyperplastic outgrowth of a primary nodule were maintained as organ cultures
for varying periods in hormone-free Medium 199. The results show that whereas noncultured nodules developed mammary tumors
at an incidence of only 15%, those passaged in organ culture gave rise to mammary tumors at an incidence of 40 to 43%. This
threefold enhancement in the oncogenicity of mammary nodules is interpreted to be due, at least in part, to a reduction in
the normal mammary cell content of nodules. Consistent with this interpretation is the observation that cultured nodules gave
rise to mammary outgrowths that were predominantly hyperplastic, whereas noncultured nodules generated outgrowths with varying
proportions of hyperplastic and normal ductal mammary tissue.
This investigation was supported by National Cancer Institute Grant CA-17862. 相似文献
18.
V. N. Katiyar J. Enami S. Nandi 《In vitro cellular & developmental biology. Plant》1978,14(9):771-774
Summary The in vitro effects of protein hormones on the stimulation of casein secretion by mouse mammary epithelial cells were studied.
Mouse mammary glands were enzymatically dissociated and used immediately or were stored frozen and thawed just before use.
Cells were cultured on floating collagen gels in the presence of insulin, cortisol and a pituitary or placental polypeptide
hormone. Casein, released into the medium, was assayed by a radioimmunoassay against one of the components of mouse casein.
Mammary cells released casein into the medium in the presence of as little as 10 ng of ovine prolactin per ml of medium. Human
growth hormone stimulated the casein secretion to the same extent as prolactin. Human placental lactogen, ovine and bovine
growth hormones were less stimulatory. Luteinizing hormone, follicle-stimulating hormone and thyroid-stimulating hormone had
no effect on the stimulation of casein secretion.
This investigation was supported by Grant No. CA 05388 awarded by the National Cancer Institute, DHEW, and by Cancer Research
Funds of the University of California. 相似文献
19.
Richard T. Glass Edwin C. Glassell Jerald J. Killion G. Mark Kollmorgen 《Cancer immunology, immunotherapy : CII》1977,2(1):57-62
Summary BDF1 mice bearing L1210 leukemia were treated with chemotherapy or in combination with neuraminidase-treated cells and BCG. Histological evaluation was done on these mice at various intervals after therapy in order to determine the rate and extent of metastatic involvement in various tissues and organs. Results were compared to tumor-bearing mice which were not treated. In all animals, tissues were classed as having minimal involvement, moderate involvement or maximal involvement based on a scale of 0 through 4. Results indicated that: (a) mice which were long term survivors did not completely reject their tumor for weeks after treatment with chemotherapy and immunotherapy; (b) complete tumor rejection did not indicate a restoration of normal tissue integrity; and (c) failure of chemotherapy-immunotherapy had no consistent pathology, but was probably due to tumor distribution rather then tumor burden per se.This study was supported, in part, by Contract No. NO1-CB-43864 and Grant No. CA 14460 from the National Cancer InstituteThe Abbreviations used are: BCNU; 1,3-bis-(2-chloroethyl)-1-nitros-ourea; Saline: 0.9% NaCl solution; BCG: Bacillus Calmette-Guerin; C. parvum: Corynebacterium parvum; pfu: plaque-forming units 相似文献
20.
Etienne Y. Lasfargues Jennie C. Lasfargues 《In vitro cellular & developmental biology. Plant》1981,17(9):805-810
Summary Out of three attempts to induce neoplasia in normal C57B1 mammary epithelial cells with the mouse mammary tumor virus (MuMTV)
only one presented signs of tumorigenicity. Immunofluorescence showed that virus synthesis took place in all three sublines
but tumorigenicity as detected by cell aggregation viability (CAV) and transplantation into syngeneic mice failed to occur
in two of them. By comparison, cells from a BALB/c spontaneous mammary tumor that do not express MuMTV were 100% tumorigenic,
whereas cells from a BALB/cfC3H tumor with a 95% virus-producing cell population had a normal CAV and were tumorigenic only
in 60% of the test animals. This lack of correlation suggested that many of the virus-producing cells were not neoplastic
and that neoplasia might occur under virus stimulation only if a restricted population of genetic cell variants existed. Accelerated
tissue culture passages of virus-free C57B1 and BALB/c normal mammary cells resulted in their spontaneous neoplasia at Passages
23 and 50 respectively; when duplicated cells cryopreserved in early passages were revived and cultivated in the same manner,
neoplasia occurred at Passages 27 and 58. The similarity of the passage numbers appears to confirm the existence of genetic
cell variants among the normal cell population.
This investigation was supported by U.S. Public Health Service Grant R01-CA-08515 from the National Cancer Institute. 相似文献