共查询到20条相似文献,搜索用时 31 毫秒
1.
Gary M. Williams Edilberto Bermudez Dominick Scaramuzzino 《In vitro cellular & developmental biology. Plant》1977,13(12):809-817
Summary The conditions for obtaining representative, adult rat hepatocyte primary cell cultures were improved such that viable yields
of 50% of the liver were produced which gave rise to cultures representing 30% of the liver. The survival of the cultures
in various media was compared revealing that in complex media, particularly containing galactose, survival was improved.
This study was supported by Contract No. N01-CP-55705 from the National Cancer Institute and Research Grant No. BC-133B from
the American Cancer Society. 相似文献
2.
P. J. Price J. I. Mansfield C. M. Hassett 《In vitro cellular & developmental biology. Plant》1979,15(2):82-85
Summary The antiviral drugs amantadine hydrochloride and rimantadine hydrochloride were tested as to their oncogenic potential using
a serial line of Fischer rat embryo cells that previously had been shown to be an accurate indicator of chemicals known to
be oncogenic in animal studies. Neither compound was found to have transforming activity. At slightly toxic levels, rimantadine
hydrochloride, but not amantadine hydrochloride, protected the same cell line from the transformation induced by the polycyclic
hydrocarbons 3-methylcholanthrene and benzo(a)pyrene.
This work was supported by Contract N01-CP-43240 within The Virus Cancer Program of the National Cancer Institute. 相似文献
3.
P. J. Price C. M. Hassett J. I. Mansfield 《In vitro cellular & developmental biology. Plant》1978,14(3):290-293
Summary Three chlorinated hydrocarbons, proposed or already in use as industrial subsitutes for the hydrocarbon trichloroethylene,
were tested for in vitro transforming potential in a Fischer rat embryo cell system (F1706), which previously has been shown
to be sensitive to transformation by chemical carcinogens. Trichloroethylene and the three substitutes (1,1,1 trichloroethane,
tetrachloroethylene and methylene chloride) all were found to induce transformation, the three substitutes being equal or
more efficient transforming agents.
This work was supported by Contract N01-CP-43240 within The Virus Cancer Program of the National Cancer Institute. 相似文献
4.
E. Y. Lasfargues W. G. Coutinho A. S. Dion 《In vitro cellular & developmental biology. Plant》1979,15(9):723-729
Summary A human breast tumor cell line BT-474 derived from an invasive ductal carcinoma was experimentally infected in vitro with
a mouse mammary tumor virus from the RIII strain (RIII-MuMTV). The virus that replicated in the human cells was characterized
as a mouse virus by immunofluorescence, electron microscopy and the presence of a specific RNA-directed DNA polymerase. The
cells themselves were human as per the karyotype and isoenzyme migration patterns. It is concluded that human cells are susceptible
to the mouse mammary tumor virus and can, eventually, support its replication.
This work was supported by USPHS Grant CA-08515 from the National Cancer Institute and by NIH Contract N01-CP-81003. 相似文献
5.
Summary Serial passage cultures of colonic epithelial cells from young rats have been maintained for more than 6 months in Eagle's
minimum essential medium buffered with HEPES (25 mM) and supplemented with 2.5% fetal bovine serum, 0.5 μg/ml insulin, 5.0 μg/ml transferrin, and antibiotics. The cells proliferated
in this medium with a population doubling time of approximately 53 h. The cells retained differentiated morphology as evidenced
by secretory activity and the presence of secretory granules, microvilli, tonofilaments, and desmosomal junctions. Further
cells at the fourth passage had normal karyotypes with 42 chromosomes and exhibited anchorage dependent growth. High concentrations
of fetal bovine serum (10 to 15%) exerted toxic effects on the colonic epithelial cell cultures.
Supported by National Cancer Institute Contract N01-CP-75914. 相似文献
6.
David M. Halton Ward D. Peterson Bharati Hukku 《In vitro cellular & developmental biology. Plant》1983,19(1):16-24
Summary Procedures that involve cell cultures require careful quality control to avoid inter- and intraspecies contamination. We have
developed and electrophoresis technique that can be used routinely in cell culture laboratories to monitor cell line integrity.
The method involves the isoenzymatic separation of nine polymorphic enzymes, three of which can be used for cell line species
determinations and seven of which can be used for human cell line characterizations. Examples of how the system has been applied
to both inter- and intraspecies identifications are described. The routine application of this protocol would be a valuable
asset for laboratories concerned with establishing effective cell culture quality control.
This work was supported by Contract N01-CP-9-1003 from the National Cancer Institute, Bethesda, MD. 相似文献
7.
Leonard J. Schiff Steven J. Moore 《In vitro cellular & developmental biology. Plant》1980,16(10):893-906
Summary A system for maintaining adult rat colonic mucosa in organ culture for up to 28 days is described. Distal colonic mucosa physically
separated from the muscle layers was cultured at 37°C on a substrate of human fibrin foam in HEPES- and bicarbonate-buffered
Waymouth's MB 752/1 medium supplemented with 10% fetal bovine serum,l-glutamine, bovine albumin, ascorbic acid, hydrocortisone, insulin, and ferrous sulfate; the optimal atmosphere for culture
was 95% O2 and 5% CO2. Viability of explants was demonstrated by tissue morphology with light microscopy, incorporation of [3H]thymidine and [3H]leucine into DNA and protein, [14C]glucosamine and [3H]fucose incorporation, and glycoprotein synthesis. Two days after initiation of culture, degeneration of surface and crypt
cells was observed. Secreted mucosubstances covered the explants. Explants maintained in 95% O2 retained a variable number of glandular crypts with normal columnar epithelium for 14 to 21 days in culture. At 28 days,
explants contained a single layer of cuboidal surface epithelium and a rare cryptlike gland.
This work was supported by the National Cancer Institute Contract N01-CP-75953 and in part by the International Cancer Research
Data Bank Program of the National Cancer Institute, National Institutes of Health, under Contract N01-CO-65341 with the International
Union Against Cancer. 相似文献
8.
Summary The effect of variations in culture conditions on survival of fragments of mouse and rat descending colon in organ culture
was studied by morphological and functional criteria. A combination of conditions demonstrated to be beneficial permitted
maintenance for at least 35 days. These included: a gaseous environment of 95% O2:5% CO2, an attachment matrix consisting of a Metricel GA-4 membrane (pore size, 0.8 μ), intermittent exposure to the gas and fluid
phases by rocking in 5 ml medium and supplementation of the medium with 1.0 μM dexamethasone and 10% FBS. During this time,
the crypt structure of the mucosal epithelium was well preserved, and DNA synthesis in the crypts and mucin production in
the crypts and superficial epithelium continued. In addition, the synthetic hormone, pentagastrin, stimulated DNA synthesis
in the mucosal epithelium of mouse colon fragment in short-term organ culture.
This work was supported by Environmental Protection Agency Grant R803998-01-1 and National Cancer Institute Contract N01-CP-75952. 相似文献
9.
Abla A. Creasey Helene S. Smith Adeline J. Hackett Kimie Fukuyama William L. Epstein Stewart H. Madin 《In vitro cellular & developmental biology. Plant》1979,15(5):342-350
Summary Three human melanoma cell lines derived from one primary and two metastatic tumors from three different patients were characterized
for growth properties usually associated with malignant transformation; these include cell morphology, growth rate, saturation
density, growth in semisolid media, colony-forming ability on contact-inhibited monolayers of normal fibroblasts and epithelial
cells, and tumorigenicity in immunosuppressed mice. Variations in expression of aberrant properties were evident among the
lines. One of the metastatic lines satisfied all the parameters of malignancy tested and the other showed a number of these
properties, whereas the primary essentially fulfilled only one. These results suggest that cultured melanoma cells reflect
the clinical variability often observed among melanoma patients and the metastatic melanoma seems to display a higher degree
of malignant transformation than the primary.
THis work was supported in part by USPHS Grant No. 5 T01 AI00332-06 from the National Institutes of Health, Contract E73-2001-N01-CP-3-3237
from the Virus Cancer Program of the National Cancer Institute, and USPHS Grant No. 0H00714-02 from the National Institute
for Occupational Safety and Health. 相似文献
10.
D. L. Fine L. O. Arthur L. J. T. Young 《In vitro cellular & developmental biology. Plant》1976,12(10):693-701
Summary Several cell culture factors were found to influence in vitro expression of mouse mammary tumor virus (MMTV) in the mouse
adenocarcinoma cell line Mm5mt/c1. Cells were propagated in a variety of commercially available cell culture media to which dexamethasone (DXM) was added as
a stimulator of MMTV production. Culture seeding density, culture medium type, and glucose concentration each influenced MMTV
production when expressed on a per cell basis. Maximum cell growth occurred in cultures grown in RPMI-1640 medium containing
insulin. Those media which provided good cell growth were not necessarily optimal for virus expression. Addition of insulin
did not stimulate MMTV synthesis although dexamethasone alone was stimulatory in all media used; however, maximum MMTV expression
was obtained when dexamethasone and insulin were used in concert. Equivalent levels of MMTV-specific cell membrane antigen,
MMTV-specific protein, and virus particles were produced at incubation temperatures of 32°, 34° or 37° C; however, higher
levels of virus-related RNA-dependent DNA polymerase (RDDP) activity were recovered from cultures incubated at 32° and 34°
C than at 37° C. Decreased levels of RDDP were attributed to enzyme thermolability at 37° C incubation.
Research sponsored by the National Cancer Institute under Contract No. N01-CO-25423 with Litton Bionetics, Inc., and Contract
No. N01-CP-33253 with the University of California. 相似文献
11.
M. Manousos M. Ahmed C. Torchio J. Wolff G. Shibley R. Stephens S. Mayyasi 《In vitro cellular & developmental biology. Plant》1980,16(6):507-515
Summary Studies conducted with virus-infected monolayer cell cultures have demonstrated the feasibility of producing several tumor-associated
viruses in microcarrier (mc) cultures (Sephadex G50 beads treated with DEAE-chloride). The efficiency of cell adherence to
mc varied with the cell type, the pH of the growth medium, and the stirring force applied to keep the mc in suspension. Most
cells attached firmly to the mc and could not be removed easily with routine trypsinization procedures. Techniques using Enzar-T
and Pronase were effective in detaching cells from mc in 10 to 15 min while retaining 95% cell viability. After detachment,
Ficoll gradients were used for rapid and complete separation of viable cell suspensions from the mc. Retrovirus production
in large volumes of mc cultures was investigated with periodic harvesting of growth fluids. Physical, biochemical, and biological
properties of the Mason-Pfizer monkey virus and the RD114 virus recovered from the mc cultures were identical to those produced
in conventional cultures. The utilization of mc has several applications in research and short-term cultures, but the as-yet-unsolved
technical problems met were found to be serious limitations when attempting mass cell culturing on a long-term basis.
For reprint requests address: Dr. Keith Jensen, Pfizer, Inc., Groton, CT 06340.
This work was supported in part under contract N01-CP-33234 within the Virus Cancer Program of the National Cancer Institute. 相似文献
12.
D. L. Klein W. Brown P. Cote W. Garner J. W. Pearson 《Cancer immunology, immunotherapy : CII》1978,4(4):229-237
Summary Immunostimulators such as Corynebacterium parvum (C. parvum), Bacillus Calmette-Guerin (BCG), pyran copolymer, and glucan were examined in the guinea pig L
2
C lymphoblastic leukemia model to determine their capacity for therapeutic modulation of the immune response of the host toward controlling leukemic cell proliferation. The dose, route, and frequency of administration of the stimulators were also evaluated as a function of time in order to obtain an optimal antileukemic effect. Results indicated that only C. parvum and BCG were capable of significantly increasing host survival when given 1 day after an inoculation of 1.5×10
4
viable leukemic cells. Administration of BCG or C. parvum, alone or in combination with irradiated blast cells on either days 4 or 7, was totally ineffective in prolonging survival. In the majority of cases, enhanced leukemic growth was observed on these days. The combination of BCG and/or C. parvum with irradiated syngeneic blast cells given 24 h after leukemia inoculation promoted a synergistic response with a significant increase in median survival time and a number of long-term survivors.This work was supported by contract N01-CP-53566 within the Virus Cancer Program of the National Cancer Institute 相似文献
13.
Normal murine melanocytes in culture 总被引:11,自引:0,他引:11
Akihiko Tamura Ruth Halaban Gisela Moellmann Janet M. Cowan Michael R. Lerner Aaron B. Lerner 《In vitro cellular & developmental biology. Plant》1987,23(7):519-522
Summary A major obstacle to applying the techniques of molecular biology to the genetics and cell biology of pigmentation has been
our inability to grow normal murine melanocytes in culture. We report here the establishment and characterization of continuously
proliferating cultures of cutaneous pigment cells from seven strains of mice. Melanocytes were grown from the dermis of newborn
mice in medium containing 12-0-tetradecanoyl-13-phorbol-acetate; a substance, such as melanotropin, that raises intracellular
levels of cyclic AMP; and an extract made from human placenta.
This work was supported by Grant R01 CA04679 from the U.S. National Institutes of Health and a fellowship to Dr. A. Tamura
from Mr. and Mrs. Allen Locklin. The chromosome studies were carried out in the laboratory of Dr. Uta Francke, Department
of Human Genetics, Yale University. JCM was supported by NIH contract number N01-CP-21037. 相似文献
14.
Judy M. Strum Elizabeth A. Hillman 《In vitro cellular & developmental biology. Plant》1981,17(1):33-43
Summary Normal breast tissue from a 17-year-old girl was grown in organ culture for 3 weeks. A comparison was made between the effects
on the epithelium of a defined culture medium containing various combinations of hormones and a serum-supplemented medium
that has been used to successfully maintain other human tissues for 4 months routinely, and in some cases for up to 1 year.
After culture for 3 weeks the explants were exposed to [3H]thymidine and autoradiographs were prepared and evaluated in order to determine labeling indexes. The only serum-free defined
medium that permitted any significant survival or labeling of the cells contained insulin + hydroxycortisone + prolactin.
However, serum-supplemented medium alone gave an even higher labeling index, and this was elevated more in media containing
either progesterone or other combinations of hormones. Our study indicates that normal human breast (removed at the early
postovulatory stage of the menstrual cycle) can be maintained in a differentiated state for 12 days in serum-supplemented
media. By 2 weeks the cells had begun to migrate onto the surface of the explant. They then began to accumulate tonofilaments
so that after 3 weeks in culture nearly all of the cells contained tonofilaments. The one exception was found in breast tissue
cultured in the presence of human chorionic gonadotropin, where the cells maintained differentiated characteristics, despite
the fact that they contained many lysosomes.
This work was supported by Grant R01 CA20764 and in part by Contract N01-CP-95640 awarded by the National Cancer Institute. 相似文献
15.
Dharam P. Chopra Lee J. Wilkoff 《In vitro cellular & developmental biology. Plant》1977,13(4):260-267
Summary In an effort to establish a test system to examine the carcinogenic potential of chemicals, mouse prostate explants were maintained
as organ cultures and the effects of carcinogenic and noncarcinogenic compounds were examined at various intervals after treatment.
The degree of hyperplasia produced by a compound was determined by the colcemid metaphase arrest technique. Extensive hyperplasia
of the prostatic epithelium occurred at 8 days after treatment with 3-methylcholanthrene, the 11–12 epoxide of methylcholanthrene,
benzo(a)pyrene and N-methyl-N-nitro-N-nitrosoguanidine. At 12 days most carcinogentreated explants were anaplastic. The noncarcinogenic
compounds, pyrene and phenanthrene, did not produce a mitotic stimulatory effect on the epithelium of the explants. The data
suggest that the organ culture system of mouse prostate may be employed as a test system to obtain preliminary information
regarding the carcinogenicity of a compound.
This investigation was supported by Contract No. NO1-CP-22064 from the Lung Cancer Segment, Division of Cancer Cause and Prevention,
National Cancer Institute, NIH, Department of Health, Education and Welfare. 相似文献
16.
Rajendra G. Mehta Leonard J. Schiff Steven J. Moore Ann Marie Buckley Marcia I. Dawson 《In vitro cellular & developmental biology. Plant》1986,22(3):164-168
Summary The mechanism of action of retinoid in reversing keratinization in hamster trachea is yet unknown. The purpose of this study
was to determine if cellular retinoic acid binding protein (CRABP) is present in tracheal epithelium following incubation
in serum-free, vitamin A-deficient culture medium for 10 days, and if the effectiveness of a retinoid in reversing keratinization
in organ culture is correlated with its ability to compete for CRABP sites. The cytosol prepared from tracheal cultures contained
CRABP at a concentration of 2.61 pmoles per mg protein. Of the four retinoids with carboxyl end group selected for the study,
two of the biological active retinoids competed for the CRABP sites. However, no correlation was observed between the biological
activity of the inactive retinoids and their ability to associate with the CRABP sites. These results indicate that even though
the action of retinoid may be mediated by retinoid binding protein, it cannot be used as a sole predicator of retinoid response
in hamster trachea.
This investigation was supported by Contract N01-CP-31012 and U. S. P. H. Grants CA30512 and CA32428, which were awarded by
the Division of Cancer Etiology, National Cancer Institute, DHHS.
Editor's Statement Tracheal organ cultures provide a useful model for the study of epithelial differentiation and carcinogenesis.
Much attention has been given to the action of retinoids in this process. Mehta et al. demonstrate a lack of correlation between
biological activity and specific cytosolic binding of members of this class of compounds, pointing out the need for a more
complete biochemical understanding of the mechanism of action and active forms of retinoids in this and other systems in vivo
and in vitro. David W. Barnes 相似文献
17.
L. F. Liebes V. M. Maher P. Scherr J. J. McCormick 《In vitro cellular & developmental biology. Plant》1976,12(3):265-268
Summary The use and construction of an automatic gas tank switching system are described. This device monitors the gas pressure in
a CO2 incubator gas system and automatically switches to a reserve tank when the main supply tank is depleted. The unit contains
an alarm system that signals either a loss of power or of gas pressure in the supply system.
This research was supported by National Cancer Institute Contract No 1-CP-33226, grants CA 13058 and CA 14680 and an institutional
grant to the Michigan Cancer Foundation by the United Foundation of Greater Detroit. 相似文献
18.
David K. Howard Jeffrey Schlom Paul B. Fisher 《In vitro cellular & developmental biology. Plant》1983,19(1):58-66
Summary We have studied the process of mammary cell transformation in vitro using a single cell clone (Clone 18) from a presumptive
epithelial cell line, C57MG, derived from a normal mammary gland; a mouse mammary tumor virus (MMTV) host-range variant (RIII)vp4;
and the potent initiating carcinogen 7,12-dimethylbenz(a)anthracene (DMBA). After several serial subcultures, cells treated with virus and then with carcinogen exhibited an altered
(transformed) morphology, a dramatic increase in anchorage independence, an increase in multinucleation after exposure to
cytochalasin B, an enhanced ability to proliferate in low Ca2+ (0.01 mM) medium, and tumorigenicity when inoculated subcutaneously into athymic (nude) mice. Although some of these phenotypic alterations
were observed also in cultures treated singly with MMTV or DMBA and in cultures exposed to DMBA before infection with MMTV,
enhanced cytochalasin B multinucleation and tumorigenicity were properties observed only in mass cultures of cloned cells
first infected with MMTV and then exposed to DMBA. This demonstrates for the first time that exposure of presumptive mammary
epithelial cells to MMTV followed by DMBA, but not to either agent alone or to DMBA followed by MMTV, results in malignant
transformation of these cells.
Support for these studies was provided in part by the National Cancer Institute, Bethesda, Md., Contract N01-CP-01018. 相似文献
19.
Several complex nutrient media were compared for their effectiveness in maintaining viable and functional mouse colon mucosa in organ culture. The order of superiority for preserving survival of normal tissues for 14 days was: Williams' Medium E > Morgan's 199 > CMRL-1066 > Waymouth's MB 752/1 > Eagle's MEM > Trowell's T8. The 3H-thymidine labeling index was highest in colon explants maintained in Morgan's 199 > Williams' Medium E > Waymouth's MB 752/1 > CMRL-1066 > Eagle's MEM. However, the very high labeling produced by Morgan's 199 medium was abnormal in comparison to in vivo levels. Supplementation with 1.0 µM dexamethasone almost always improved crypt survival and maintained normal DNA synthetic activity.Supported by National Cancer Institute contract No. 1-CP-75952 and grant No. CA-29602. 相似文献
20.
Paul J. Price Angela E. Auletta Martin P. King Patricia M. Hugunin Robert J. Huebner 《In vitro cellular & developmental biology. Plant》1976,12(8):595-598
Summary Fischer rat embryo cells chronically infected with Rauscher murine leukemia virus, and known to be sensitive to transformation
by potent chemical carcinogens, were transformed by the weak carcinogen 4-nitropyridine-1-oxide. Transformed cells grew in
semi-solid agar and produced tumors in newborn Fischer rats. Transformation was inhibited by antisera specific for the ecotropic
Rauscher murine leukemia virus, but not by antisera of equal toxicity specific for xenotropic Swiss mouse AT-124 virus.
This work was supported by contract NO1-CP-43240 within the Virus Cancer Program of the National Cancer Institute. 相似文献