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1.
Arthur E. Greene Jesse Charney Warren W. Nichols Lewis L. Coriell 《In vitro cellular & developmental biology. Plant》1972,7(5):313-322
Summary Three mosquito cell cultures designated as Suitor's clone ofAedes aegypti, Culiseta inornata, andAedes vexans were shown to be moth by immunological, karyological, and isozyme analyses. The cells reacted with rabbit antimoth serum
but not rabbit antimosquito serum. Chromosome analyses indicated Lepidopteran rather than Dipteran morphology, and three isozyme
systems were confirmative. Any one of these assays would be sufficient to indicate that contamination had occurred and could
be used as a periodic check for identity of cell cultures. Morphology and growth characteristics are also valid criteria to
distinguish between these particular orders of insect cells.
These studies were supported by Grant CA-04953-12 from the National Cancer Institute; General Research Support Grant FR-5582
from the National Institutes of Health; and Grant-in-Aid Contract M-43 from the State of New Jersey.
Recipient of Research Career Award 5-K3-16,749 from the National Institutes of Health. 相似文献
2.
Arthur E. Greene Jesse Charney Warren W. Nichols Lewis L. Coriell 《In vitro cellular & developmental biology. Plant》1972,7(6):313-322
Summary Three mosquito cell cultures designated as Suitor's clone ofAedes aegypti, Culiseta inornata andAedes vexans were shown to be moth by immunological, karyological, and isozyme analyses. The cells reacted with rabbit antimoth serum
but not rabbit antimosquito serum. Chromosome analyses indicated Lepidopteran rather than Dipteran morphology, and three isozyme
systems were confirmative. Any one of these assays would be sufficient to indicate that contamination had occurred and could
be used as a periodic check for identity of cell cultures. Morphology and growth characteristics are also valid criteria to
distinguish between these particular orders of insect cells.
These studies were supported by Grant CA-04953-12 from the National Cancer Institute; General Research Support Grant FR-5582
from the National Institues of Health; and Grant-in-Aid Contract M-43 from the State of New Jersey.
Recipient of Research Career Award 5-K3-16,749. from the National Institutes of Health. 相似文献
3.
Gerard J. McGarrity Lewis L. Coriell 《In vitro cellular & developmental biology. Plant》1973,9(1):17-18
Summary A commercially available anaerobic generator and incubation system that develops a low oxidation-reduction potential was used
for the assay of cell cultures for mycoplasmal contamination. Mycoplasma broth and agar media supplemented with dextrose,
yeast extract, and horse serum were used. This system supported growth of some mycoplasmas that failed to grow in incubators
with 5% CO2 in nitrogen previously used in culture of mycoplasmas in this laboratory.
This work was supported by a grant from the John A. Hartford Foundation General Research Support Grant No. 5 SO1 RR05582-4
from the National Institutes of Health, and by a Grant-in-Aid Contract from the State of New Jersey. 相似文献
4.
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
5.
Joseph L. Melnick Janet S. Butel Satvir S. Tevethia Nilambar Biswal Matilda Benyesh-Melnick 《In vitro cellular & developmental biology. Plant》1971,6(5):349-354
Summary This paper describes some current work pertaining to transformation of cells by oncogenic viruses.
Part I includes: (1) the effect of a deoxyribonucleic acid (DNA) tumor virus (SV40) on the antigenic characteristics of transformed
cells; (2) in vitro and in vivo methods of detecting virus-specific surface antigens; (3) the role that the host cell may
play in the expression of virus-coded antigens; and (4) the presence of virus-induced antigens as a possible mechanism of
the apparent nononcogenicity of certain virus variants.
Part II discusses (1) the physicochemical properties of the nucleic acid of a ribonucleic acid (RNA) tumor virus-the Moloney
sarcoma-leukemia virus (MSV-MLV) complex —(2) a preliminary analysis of viral RNA replication in cells transformed by MSV-MLV,
and (3) application to human tumors.
Supported in part by Research Grant CA 04600 and by Research Contract PH 43-68-678 within the Special Virus-Cancer Program,
National Cancer Institute, National Institutes of Health.
Recipient of Research Career Development Award 5-K3-CA 38,614 from the National Cancer Institute, National Institutes of Health 相似文献
6.
Gerard J. McGarrity 《In vitro cellular & developmental biology. Plant》1976,12(9):643-648
Summary Environmental sampling was performed during trypsinization and passage of 3T-6 cell cultures that contained a mean of 4.3×107 colony forming units (CFU) per ml supernatant ofA. laidlawii. The lip of the culture flask and the outside of the used pipet were always heavily contaminated. The outside of the culture
flask (3/7), the work surface (8/12) and the outside of a pan of disinfectant (4/5) were regularly contaminated with mycoplasmas.
Airborne mycoplasmas were detected eight of 32 times (25%) by settling plates; simultaneous forced-air samplers by two different
methods were always negative. The technician’s hands were contaminated two of 15 samples. When hands were contaminated, more
contamination was detected in the environment. Droplets ofA. laidlawii andM. orale inoculated onto work surfaces survived drying for a minimum of 3 days, even in laminar airflow cabinets. Twenty-five of 31
(80.6%) cell culture technicians carriedM. salivarium in their throats; only two carriedM. orale. It is concluded that mycoplasma-infected cultures are the most common source of further infection. Recommendations for prevention
and control of mycoplasmal infection are listed.
These studies were supported in part by Contract No. 1-GM-2112 from the National Institute of General Medical Sciences, Contract
No. 1-CB-23868 from the National Cancer Institute, General Research Support Grant 5-S01-RRO5582 from Research Resources, National
Institutes of Health, and by a Grant-in-Aid from the State of New Jersey. 相似文献
7.
Summary Bovine herpesvirus-1 infection in hamster embryo cells was found to be dependent upon input multiplicity; productive infection
was achieved at input multiplicities greater than one, while persistent infection was established when input multiplicities
were about 0.5. This persistence was characterized by a noncyclic, minimal degree of cytopathic effect with a low level of
released virus. Maintenance of the persistently infected cultures did not require external supportive measures. Subcultivation
of the persistently infected cultures led to virus replication followed by CPE and then cell regrowth. Within 3 to 4 weeks
after subcultivation a persistent infection was re-established. The possible mechanism for the bovine herpesvirus persistence
in hamster cells is discussed.
This work was supported in part by Public Health Service Research Contract FDA 233-74-1035 and by Research Grant AI-08648
from the National Institute of Allergy and Infectious Diseases, National Institutes of Health. 相似文献
8.
Gerard J. McGarrity Lewis L. Coriell 《In vitro cellular & developmental biology. Plant》1971,6(4):257-265
Summary The most frequent causes of cell culture contamination are poor techniques and housekeeping and inadequate sterility testing
of supplies and culture media. The most common contaminants of cell cultures areMycoplasma, Torula sp., andPseudomonas sp. Routine procedures used in this laboratory to control or prevent accidental contamination include the use of filtered
laminar air flow in transfer rooms, elimination of antibiotics wherever possible, careful execution of aseptic procedures,
periodic review and discussion of techniques, sterility testing of all media components, use of approved clothing, and effective
cleaning and disinfection procedures.
These studies were supported by a grant from the John A. Hartford Foundation General Research Support Grant FR-5582 from the
National Institutes of Health, Grant in Aid Contract M-43 from the State of New Jersey, and by Grant CA-04953-11 from the
National Cancer Institute. 相似文献
9.
Initiation and characterization of a diploid cell line from larval tissues ofAedes dorsalis (Meigen)
Barbara E. Cahoon James L. Hardy William C. Reeves 《In vitro cellular & developmental biology. Plant》1978,14(3):255-260
Summary Mosquito cell cultures were initiated from the minced tissues of newly hatchedAedes dorsalis (Meigen) larvae. Continuous cell division occurred only after an adaptive period of approximately 6 months. Optimal growth
of the cells required a relatively low pH of 6.5. Karyological studies showed that the cells have remained diploid (2n=6)
for 60 serial passages and that the cultures are free of contaminating cells. The cultures also were shown to be free of bacteria
(includingMycoplasma), fungi and virions. Subpopulations (strains) of the original parental cultures have been selected and characterized on the
basis of morphology, karyology, growth rate and monolayer formation.
These studies were supported in part by funds from the Office of Naval Research, by Research Grant AI03028 from the National
Institute of Allergy and Infectious Diseases, and by General Research Support Grant I-SO1-FR-05441 from the National Institutes
of Health, U.S. Department of Health, Education and Welfare. 相似文献
10.
Lawrence W. Anderson Keith G. Danielson Howard L. Hosick 《In vitro cellular & developmental biology. Plant》1979,15(11):841-843
Summary A cell line and subline with epithelial characteristics were established from mouse mammary hyperplastic alveolar nodules
(HAN). The cells do not grow in suspension cultures in vitro or form tumors in vivo. The cells do produce significant amounts
of C-type and A-type virus and low amounts of plasminogen activator.
This work was supported by ACS Postdoctoral Fellowship PF-1473, and Grant 5-ROI-CA-16392-05 and Contract NO1-CB-63986 from
the National Institutes of Health. 相似文献
11.
A method for the continuous cultivation of mammary epithelium 总被引:1,自引:0,他引:1
Etienne Y. Lasfargues Dan H. Moore 《In vitro cellular & developmental biology. Plant》1971,7(1):21-25
Summary Established cell lines of mammary epithelium have been obtained from mice, rats, and hamsters. Maintenance and replication
of the epithelium in serial subcultures were dependent on their periodic treatment with collagenase. Because collagenase is
not cytotoxic and has maximum efficiency at neutral pH in isotonic saline solutions containing calcium and magnesium, this
enzyme can be introduced directly into the culture medium; cells have been maintained for 3 days in such a medium with serous
enrichment at no detriment to them. Up to 10% concentrations of serum have not interfered with enzymatic activity.
Supported by United States Public Health Service Grant CA-08515 and CA-08740 from the National Cancer Institute. General Research
Support Grant FR-5582 from the National Institutes of Health, and Grant-in-Aid Contract M-43 from the State of New Jersey. 相似文献
12.
Michael G. Gabridge Robert B. Polisky 《In vitro cellular & developmental biology. Plant》1977,13(8):510-516
Summary The amount of adenosine triphosphate (ATP) in hamster trachea organ cultures was determined with a technique based on light
emission from a luciferin/luciferase/ATP reaction. The amount of ATP, expressed as ng per mg dry weight, was consistent in
tracheal explants prepared from various animals and changed negligibly when explants were cultivated in vitro for several
days. The amount of ATP was related directly to cellular activity and integrity in the epithelium since inactivation by heat
or freeze-thaw rapidly depleted measurable ATP, and ciliary activity and ATP content were related directly. When tracheal
explants were infected with 105 to 107 CFU of virulentMycoplasma pneumoniae cells, both ciliary activity and ATP content in the tissue dropped dramatically after approximately 5 to 8 days (up to 85%
and 60% decreases, respectively). Exposure of explants to 50 to 200 μg per ml of purifiedM. pneumoniae membranes also caused significant decreases in ciliary activity and ATP. When explants were infected with attenuated or nonvirulent
mycoplasmas, ciliary activity was only slightly decreased, while ATP values often rose slightly. The technology associated
with the determination of ATP levels in tracheal explants should prove useful as a new, objective, analytical approach to
cell viability in organ cultures.
This investigation was supported in part by the National Institutes of Health (PHS Grant AI 12559), by a Biomedical Sciences
Support Grant made to the University of Illinois School of Life Sciences, and by the University Research Board. 相似文献
13.
Summary A method is described for obtaining and culturing large numbers of lung cells from normal adult male rats. The lungs were
perfused in situ to remove blood cells and then perfused via the trachea with a trypsin-collagenase solution to initiate tissue
digestion. The tissue was further digested in the enzyme solution and approximately 2×108 viable lung cells were obtained per animal. Primary cultures contained a mixed cell population. Through eight subcultures
about 70% of the cell population possessed an epithelial-like morphology, whereas the remaining 30% was fibroblast-like. Three
clones of epithelial-like cells were isolated at the fourth subculture. The mass culture lung cells and the epithelial-like
clone that was studied retained a normal karyotype and did not grow in soft agar. Both the mass culture cells and the epithelial
clone metabolized the lung carcinogen benzo(a)pyrene (BP) to water-soluble products. Furthermore, the mass culture lung cells
metabolized BP to intermediate(s) which mutated Chinese hamster V79 cells from ouabain sensitivity to ouabain resistance.
These lung cell cultures have potential use in cell transformation, mutation and carcinogen metabolism studies.
Visiting scientist from Hungary.
This research was supported by Grant 5 R01 CA20022 and Public Health Service Contract N01 CP33278 from the Division of Cancer
Cause and Prevention, National Cancer Institute, National Institutes of Health. 相似文献
14.
Kenneth J. Lazarus George A. Hashim Philip Y. Paterson Eugene D. Day 《Neurochemical research》1984,9(9):1295-1308
Three antisera to myelin basic protein—a rabbit antiserum pool against rat myelin, a rabbit antiserum pool against rat myelin basic protein (MBP), and a monkey antiserum against bovine MBP—were found to contain detectable levels of antibodies that would bind radiolabeled S49 (GSLPQKAQRPQDENG). Strongly encephalitogenic in Lewis rat, S49 is a synthetic peptide representing residues 69–84 of bovine MBP with a deletion of glycine-76 and histidine-77 to make it analogous to rat and guinea pig MBPs. The rabbit antimyelin antiserum and the monkey anti-MBP antiserum contained antibodies directed against a non-sequential determinant that required asparagine 84, the glycine-histidine deletion, and residues 69–71 for maximal activity. S49-reactive antibodies from the rabbit anti-MBP antiserum were directed solely against a sequential determinant comprising residues 69–71. S49-reactive antibodies from all three antisera reacted in liquid phase with purified intact rat, guinea pig, and bovine MBP showing that the determinant is exposed for B cell recognition even in bovine MBP and can serve both as immunogen and reactant.This work supported at Duke University Medical Center by Research Grant NS-10237 from the National Institutes of Health of the U.S. Public Health Service and the Medical Scientist Training Program Grant #5-T32-OMO-7171-08; at St. Luke's Hospital Center by NS-15322 from the National Institutes of Health of the U.S. Public Health Service; and at Northwestern University by Research Grant NS-06262 from the National Institutes of Health of the U.S. Public Health Service. 相似文献
15.
The objective of this study was to determine if thermophilic fungi exist in the mycoflora of man and in the aeroflora of his environment.Humicola lanuginosa andHumicola grisea were isolated from 5 of 55 samples of outside air. Three thousand cultures were taken from the nasal mucosae, skin surfaces and recta of 570 children. Cultures were incubated at 50°C. Thermophilic fungi were isolated from 6 of 287 children receiving immunosuppressive therapy for malignancies and from 1 of 283 normal children.H. lanuginosa was recovered from the skin of one, the rectum of one and the nasal mucosae of three patients.Mucor pusillus was isolated from the nasopharynges of two patients.Further studies are now indicated to determine the pathogenicity of these organisms with respect to tissue invasive disease, antigenicity and metabolite toxicity.Supported by General Research Support Grant RR-05584 from National Institutes of Health; Cancer Research Center Grant CA-08480 and Training Grant CA-05176 from the National Cancer Institute, National Institutes of Health and by ALSAC. 相似文献
16.
Georgirene D. Vladutiu Richard M. Fike Valerie T. Amigone 《In vitro cellular & developmental biology. Plant》1981,17(7):588-592
Summary Fibroblasts derived from patients with I-cell disease have been shown to accumulate many natural substrates including a three
to fourfold increase in sialic acid content compared to that found in normal fibroblasts. This diverse accumulation of storage
material is due to a massive deficiency of multiple lysosomal hydrolases as they are preferentially excreted into the culture
fluid. There is evidence that the I-cell plasma membrane itself is abnormal with respect to certain transferase activities
and in its sensitivity to freezing and Triton X-100. In this study, we have shown that a neuraminidase-sensitive substrate,
and perhaps others in I-cell fibroblasts, contribute to an increased electronegativity of the I-cell fibroblast surface and
to the cells' sensitivity to freezing. We also found that neuraminidase treatment of I-cell fibroblasts before preservative
freezing in liquid nitrogen enables the cells to adapt more easily to subculture upon thawing.
This project was supported in part by National Institutes of Health (NIH) BRSG Grant RR-05493, NIH Grant 1-R01-HD-11453-01-A1,
National Science Foundation Grant PCM 77-05733, and Maternal and Child Health Service Project 417. Georgirene D. Vladutiu
is the recipient of Research Career Development Award 1K04 HD 00312-01A1 from the National Institutes of Health. 相似文献
17.
C. W. Long R. DelGiudice R. S. Gardella M. Hatanaka 《In vitro cellular & developmental biology. Plant》1977,13(7):429-433
Summary Human H. Ep-2 and mouse 3T6 cells infected byMycoplasma hyorhinis showed an increase in [3H]uracil uptake and a more than 20-fold increase in the activity of uracil phosphoribosyltransferase (UraPRT). Uninfected
cell cultures gave background levels of this enzyme activity. A survey of 16 strains of mycoplasma showed 13 to possess UraPRT
activity. Rabbit kidney cells (RK13) were infected with eight different strains of four mycoplasma species known to be common
cell culture contaminants. Seven of the eight cell cultures showed elevated UraPRT activities four days after infection. This
enzyme activity may be of value in monitoring cell cultures for mycoplasma and aid in classification.
This work was supported by Contract NO1-CP-53530 with the National Cancer Institute, National Institutes of Health, and Contract
FDA 74-41 of the Food and Drug Administration, Bethesda, Maryland 20014. 相似文献
18.
Human semen as a source of epithelial cells for culture 总被引:3,自引:0,他引:3
Stephanie Gordon Phillips David M. Phillips Elvin A. Kabat Orlando J. Miller 《In vitro cellular & developmental biology. Plant》1978,14(8):639-650
Summary When washed cells from human semen samples were plated out, epithelial cultures were obtained. The human ejaculates used as
starting material contained, in addition to spermatazoa, 103 to 107 cells of other types, including granulocytes, macrophages lymphocytes, spermatocytes and epithelial cells. Although no fractionation
of cell types was attempted, semen samples yielded epithelial cultures uncontaminated by fibroblasts. The cultured cells appeared
characteristically epithelial with a polygonal shape, interdigitating cell membranes, and desmosomes. ABH blood-group antigenic
determinants of the donor were expressed with variable frequency as a surface antigen on these cells. About half the trials
gave some cell attachment. Most cultures remained as small, tight colonies, but a few reached confluency in about 5 weeks
and could be subcultured successfully. Cell proliferation, as monitored by [3H]thymidine incorporation into nuclear macromolecules, ceased in less than 2 months.
Aided by grants from the National Science Foundation (BMS-72-02219 A04 and PCM 76-81029 to E. A. K.), the Public Health Service
(CA 12504 to O. J. M.), and the National Foundation-March of Dimes. The earlier portions of this study were carried out under
a Program Project Grant from the National Institutes of Health (No. 5 PO GM 18153). 相似文献
19.
The genetic rescue of Pgd
n
lethal alleles, accomplished by combining them with mutations lacking glucose-6-phosphate dehydrogenase activity, has led to the hypothesis that Pgd
n
lethality may be due to the accumulation of 6-phosphogluconate. In this article we report the rescue of Pgd
n
/Y males by dietary supplements (fructose and linolenate) designed to minimize 6-phosphogluconate production.This investigation was supported by Research Grant GM-15691 and Training Grant T01-GM-0685 of the National Institutes of Health. 相似文献
20.
Isolation,characterization, and long-term culture of fetal bovine tracheal epithelial cells 总被引:5,自引:0,他引:5
Brenda L. Schumann Terence E. Cody Marian L. Miller George D. Leikauf 《In vitro cellular & developmental biology. Plant》1988,24(3):211-216
Summary Epithelial cells were isolated from fetal bovine trachea by exposing and stripping the mucosal epithelium from the adjacent
connective tissue. The tissue was minced and enzymically dissociated in Ca-Mg-free medium containing dispase and dithiothreitol.
The stripping procedure and selective trypsinization produced epithelial cell cultures free of fibroblasts. Seeded on plastic,
the plating efficiency was 21.5% with a doubling time of 24 h. Dome formation, evidence of occluding junctions and active
ion transport characteristic of epithelial cells, was common. Growth of the cells on glass, collagen, and Engelbreth-Holm-Swarm
(EHS) substrate demonstrated a striking difference in morphology. Cells grown on EHS presented a more distinctly three-dimensional
growth pattern and many more microvilli when compared to cells grown on glass or collagen. The cells retained their epithelioid
characteristics through more than 30 passages as shown by the presence of distinct apical and basolateral membranes, tight
junctions, and positive keratin staining.
This study was supported in part by grants BRSG S07 RR05408-25, Biomedical Research Support Grant Program, Division of Research
Resources, by ES 00159, Center Grant, National Institutes of Environmental Health Sciences, by R23-HL37621, New Investigator
Award, National Heart, Lung and Blood Institutes, National Institutes of Health, and by the Health Effects Institute, an organization
jointly funded by the U.S. Environmental Protection Agency (Assistance Agreement X-8120059) and automotive manufactures. The
contents do not necessarily reflect the views of policies of HEI, EPA, or automotive manufacturers. 相似文献