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1.
Michael H. Simonian Mark L. White David A. Foggia 《In vitro cellular & developmental biology. Plant》1987,23(4):247-252
Summary The life span and growth from clonal density of bovine adrenocortical cell cultures were studied in serum-supplemented medium
and a serum-free defined medium, which supported sustained cell proliferation and steroid production. The total culture life
span was 79 population doublings in serum-supplemented medium with fibroblast growth factor (FGF) and 36 population doublings
in the defined medium without serum. Older passage cell cultures grown in the defined medium progressively lost the ability
to produce 11β- and 21-hydroxylated steroids, which was observed previously for cultures in serum-supplemented medium, and
also had a decline of 17α-hydroxylated steroid production. The cloning efficiency in the defined medium was 12.2% as compared
to 24% in serum-supplemented medium with FGF. Five isolated clonal cell lines grown in the defined medium were characterized
for steroid function in response to steroidogenic agents. All five clonal cell lines had stimulated steroid production with
8-bromo-cAMP, but only four of the clonal lines were stimulated also by adrenocorticotropin. None of the clonal cell lines
produced 11β-, 21- or 17α-hydroxylated steroids in response to treatment with either steroidogenic agent, results that were
similar to data obtained from older mass cultures. The apparent deficiency of the defined medium as compared to serum-supplemented
medium for maximum support of the culture life span and cloning efficiency may be useful in studies of cellular aging and
its relation to differentiated function for this cell culture system.
This study was supported by the Iowa Diabetes and Endocrinology Research Center (grant AM25295 from the National Institutes
of Health, Bethesda, MD). D.A.F. was supported by a National Research Service Award from the National Institutes of Health
(grant HL07485). 相似文献
2.
Joseph M. Lombardo Karl W. Lanks 《In vitro cellular & developmental biology. Plant》1982,18(3):251-253
Summary Cell lines derived from a murine neuroblastoma, Clone N18, were cured of theirM. hyorhinis infection by in vivo passage. The major variable determining success of this method was found to be the incubation time in
vivo. Infected cells maintained in vivo for 27 d or more and then placed in culture were free of mycoplasma whereas those
maintained in vivo for 7 or 14 d were found to still be infected. This approach to eliminating mycoplasma infection may be
successful using other tumor cell lines.
This work was supported in part by Grants ES01530, GM24592, and GM26167 from the National Institutes of Health, Bethesda,
MD. 相似文献
3.
Schroeder Jamie M. Chamberlain Constance Davidson Elizabeth W. 《In vitro cellular & developmental biology. Plant》1989,25(10):887-891
Summary Increasing doses ofBacillus sphaericus toxin were used to select a toxin-resistant cell line from theCulex quinquefasciatus line. This resistant cell line was proven to beC. quinquefasciatus in origin by isozyme analysis and karyotype. The resistant line bound fluorescent-labeled toxin as did the unselected susceptible
line. A high level of resistance was quickly achieved, and this level was maintained after 4 mo. culture in the absence of
toxin.
This work was supported by grant number RO1 AI 22702 from the National Institutes of Health, Bethesda, MD. 相似文献
4.
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
5.
Barbara Ruben Migeon 《Biochemical genetics》1968,1(4):305-322
Somatic hybrids of drug-resistant mutant hamster and mouse cell lines have been isolated and propagated in long-term culture and have been studied in respect to karyotype and three enzymes. During the course of propagation the long-surviving hybrid clones show progressive loss of telocentric chromosomes associated in at least one case with loss of mouse enzyme. Hybrid clones showed hybrid molecules for malate dehydrogenase (MDH), lactate dehydrogenase (LDH), and 6-phosphogluconate dehydrogenase (6PGD) made up by recombination of parental subunits.This work was supported by National Institutes of Health Grant HD 00486. 相似文献
6.
Lindegren Carl C. Lindegren Gertrude Drysdale Robert B. Hughes John P. Brenes-Pomales Arturo 《Genetica》1957,28(1):1-24
Summary A single tetrad in which one genetical marker had segregated irregularly was analyzed genetically by outcrossing each culture derived from the tetrad to other haploid clones. Regular segregation in the resultant hybrids indicated that the cultures were all haploid. The original ascus was tetratype proving that all four nuclei had survived after reduction. All clones were haploid proving that the irregularities could not have arisen from fusion following an extra mitosis. It is inferred that the extra recessive was the result of an interaction in the hybrid in which a dominant was converted into a recessive allele. The converted clone was identified by the intermediate character of its physiological activity.This work has been supported by grants from the National Cancer Institute of the National Institutes of Health, Public Health Service, Contracts C-1179 and C-2140 and Anheuser-Busch, Inc. 相似文献
7.
Grazyna Adamus Z. Suzanne Zam Scotts Emerson Paul A. Hargrave 《In vitro cellular & developmental biology. Plant》1989,25(12):1141-1146
Summary We present a practical method for the rescue of previosly stable hybridoma clones which increases the proportion of desired
cells in the population before cloning by limiting dilution. When the antibody activity of a culture supernatant was lower
than that previously obtained, a precloning distribution at a density of 10 cells per microtiter well greatly improved the
chances of obtaining a single active clone by subsequent limiting dilution. The Poisson distribution model was used to evaluate
the method. Probabilities calculated clearly demonstrate the advantage of this precloning distribution step when attempting
to isolate a hydridoma cell line that is relatively rare in a population.
This work was supported in part by grants EY 06225 and EY 06226 from the National Eye Institute of the National Institutes
of Health, Bethesda, MD and by an unrestricted departmental award from Research to Prevent Blindness, Inc. 相似文献
8.
Development of two cloned epithelial cell lines from normal adult mouse and rat ventral prostates 总被引:1,自引:0,他引:1
Yoshinobu Kubota Eugene B. Gehly Karl H. Link Charles Heidelberger 《In vitro cellular & developmental biology. Plant》1981,17(11):965-978
Summary Two epithelial cell lines were established, one from adult C3H mouse and one from adult Fischer rat ventral prostate. These
cell lines were obtained from explant cultures, using Ham's F12 medium supplemented with HEPES, insulin, testosterone, hydrocortisone,
epidermal growth factor, and 7.5% fetal bovine serum. A low concentration of trypsin and EDTA in Ca++-and Mg++-free phosphate buffer was used for passaging the cells. The rat cell line was established following implantation of prostate
tissue in nude mice. These cell lines stained positively for acid phosphatase and were dependent upon epidermal growth factor
for growth. Morphological studies, including electron microscopy, revealed a highly characteristic epithelial morphology of
both cell lines. These cell lines have hypotetraploid chromosome numbers and are capable of metabolizing benzo(a)pyrene. We propose the application of these cells as models for the study of prostate carcinogenesis.
This work was supported in part by Grant CA-21, 746, and by the Electron Microscope Core Facility on Grant CA-14,089, from
the National Cancer Institute, National Institutes of Health, Bethesda, MD. 相似文献
9.
Midori Hosobuchi Martha R. Stampfer 《In vitro cellular & developmental biology. Plant》1989,25(8):705-713
Summary Normal human mammary epithelial cells (HMEC) from different individual reduction mammoplasty specimens were all growth inhibited,
and showed a flattened, elongated morphology in response to human recombinant transforming growth factor β1 (TGFβ). The degree
of growth inhibition varied among specimens, but none of the normal HMEC maintained growth in the continued presence of TGFβ.
The degree of growth inhibition also varied with cell age in vitro, cells closer to senescence being more sensitive. TGFβ
sensitivity was additionally assayed in two established cell lines derived from one of the reduction mammoplasty specimens
after exposure to benzo(a)pyrene. Although varying degrees of growth inhibition and morphologic changes were observed in the
cell lines, both lines contained populations that maintained active growth in the presence of TGFβ. Subclones of these lines
demonstrated a great plasticity in their growth response to TGFβ, with individual clones ranging from strongly growth inhibited
to nearly unaffected. These results suggest that multiple factors influence the extent of TGFβ-induced growth effects on both
normal and transformed mammary epithelial cells, and that some of these factors may act through epigenetic mechanisms.
This work was supported by CA24844 from the National Institutes of Health, Bethesda, MD, and the Office of Energy Research,
Office of Health and Environmental Research of the U.S. Department of Energy under contract DE-AC03-76SF00098. 相似文献
10.
Jan Schwarz Gerald L. Baum C. J. K. Wang Eula L. Bingham Harold Rubel 《Mycopathologia》1957,8(3):189-193
Summary The first repeatedly successful infection withHistoplasma capsulatium of pigeons and chickens is reported.From 44 intravenously infected pigeonsHistoplasma capsulatum could be recovered by culture in 20 cases as long as 45 days after injection; from 10 chickens 5 positive cultures were obtained.This research was in part supported by grant E-576 of the National Institutes of Health. 相似文献
11.
Carmen Raventos-Suarez 《In vitro cellular & developmental biology. Plant》1985,21(3):161-164
Summary We report that synchronized cultures ofPlasmodium falciparum with up to 40% parasitized cells can be obtained with the use of low, red blood cell suspensions and only daily replacement
of culture medium. These cultures contained not only a reduced proportion of uninfected red cells but also of population of
cells with brief and equal time of exposure to culture conditions. Such high parasitemias are desirable for studies of ring-staged
parasites (for which enrichment techniques are not available) and late-staged parasites when the manipulations for enrichment
are inappropriate or unsuccessful.
This work was supported by Grant HL-21016 from the National Institutes of Health, Bethesda, MD
Preliminary observations on this report were presented at the Poster session at the Red Cell 6th Ann Arbor Conference, Michigan,
October, 1983. 相似文献
12.
Tileye Feyissa Li-Hua Zhu Legesse Negash Margareta Welander 《Plant Cell, Tissue and Organ Culture》2007,88(3):277-288
The effects of growth regulators, wounding and antibiotics on regeneration of Hagenia abyssinica were investigated and the rolB gene was introduced into this species by Agrobacterium-mediated transformation. Regeneration was affected by type of growth regulators, wounding and antibiotics. Up to 100% regeneration
could be obtained. Three transformed clones (T1, T2.1, T2.2), confirmed by PCR and Southern blot, were obtained only by excluding
kanamycin from the selection medium 6 weeks after culture, followed by selection during shoot multiplication. RT-PCR revealed
strong expression of rolB gene in shoots and roots of all the transgenic clones, but from leaf samples, it was detected only in T1. Rooting frequency
was 77% (T1), 50% (T2.1), 57% (T2.2) and 0% for control shoots on growth regulator-free rooting medium. 相似文献
13.
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
14.
William J. Lindblad Louise C. Flood 《In vitro cellular & developmental biology. Plant》1987,23(6):413-416
Summary Several dermal fibroblast lines have been established from explants taken from adult rats. The cells have been cultured for
2 yr and possess stable and well-defined growth characteristics through subculture 18. The cells are readily stored in liquid
nitrogen with good viability after thawing.
Collagenase activity secreted into the culture medium of the cells at different periods of growth has been examined. There
is an 88% drop in total enzyme activity present in the medium between 4 and 14 d of culture, when the cells were plated to
reach confluence at Day 8 to 10. A more pronounced fall is noted at earlier times when the cells are plated at a higher density.
The correlation between DNA content of the cell monolayer and enzyme activity was −0.895, indicating a possible relationship
between the growth of cells and collagenase release.
This study was supported in part by grant AM 30856 from the National Institutes of Health, Bethesda, MD. 相似文献
15.
E. A. Schwinghamer 《Antonie van Leeuwenhoek》1967,33(1):121-136
Many antibiotic-resistant mutants derived from effective strains ofRhizobium — mainlyR. leguminosarum andR. trifolii showed some loss of effectiveness on the homologous host. The frequency and degree of such changes in effectiveness depended on the type of antibiotic-resistance involved in the mutations. Resistance to streptomycin, spiramycin, chloramphenicol, or the tetracycline group of antibiotics was associated with little or no change. Resistance to D-cycloserine, novobiocin, vancomycin, bacitracin or penicillin was accompanied by partial or full loss of effectiveness in about one-half of the mutants. Full loss of effectiveness occurred most frequently among mutants selected for resistance to viomycin or neomycin. In contrast, restoration of the ability for effective symbiosis occurred in prototrophic revertants of a non-nodulating, purine-requiring mutant. Alteration of cell-wall characteristics is suggested as a possible basis for decrease of effectiveness among mutants resistant to those antibiotics, notablyD-cycloserine, which are known to inhibit cell-wall synthesis.This investigation was supported by a Public Health Service research career development award (1-K3-GM-22, 594) and research grant (GM-12131) from the National Institutes of Health. 相似文献
16.
We have examined the effect of addition of hydroxocobalamin to growth medium on the activity of the adenosylcobalamin-requiring enzyme methylmalonyl CoA mutase in normal human fibroblasts and in mutant human fibroblasts derived from patients with inherited methylmalonicacidemia. The mutant cell lines were assigned to four distinct genetic complementation groups (cbl A, cbl B, cbl C, and cbl D), each deficient in some step in the synthesis of adenosylcobalamin from hydroxocobalamin. After control cells were grown in cobalamin-supplemented medium, mutase holoenzyme activity increased markedly in a time- and concentration-dependent fashion. Growth in cobalamin-supplemented medium had no effect on mutase activity in some mutant lines belonging to the cbl B group, while activity increased severalfold in other cbl B mutants and in all cbl A, cbl C, and cbl D mutants examined, although mutase activity was still <10% of control. Comparison of mutase holoenzyme activity and total propionate pathway activity suggests that enhancement of mutase activity in mutant cells after cobalamin supplementation to values 5–10% of control may be sufficient to overcome the inherited metabolic block and to restore total pathway activity to normal.This work was supported in part by a research grant from the National Institutes of Health (AM 12579). H. F. W. is a recipient of a traineeship from the National Institutes of Health (T01-GM02299). 相似文献
17.
Dianne M. Rausch Sidney B. Simpson Jr. 《In vitro cellular & developmental biology. Plant》1988,24(3):217-222
Summary The immune suppressed lizard,Anolis carolinensis, can be used to test for in vivo tumor production by cell lines derived from a variety of ectothermic vertebrates. Cell lines
tested for tumor production were also assessed for loss of attachment-dependent proliferation and contact inhibition of cell
overlap. The results demonstrate that the criteria standardly used to assess transformation and neoplastic change in cultured
mammalian cells apply equally well to cultured cells from ectotherms.
Supported by grants AG01476 and NS24162 from the National Institutes of Health, Bethesda, MD. 相似文献
18.
Herbert Lazarus E. F. Barell S. Oppenheim Awtar Krishan 《In vitro cellular & developmental biology. Plant》1974,9(5):303-310
Summary Two lymphocytic cell lines were obtained from a single specimen of peripheral blood buffy coat from a patient with acute lymphoblastic
leukemia who concurrently developed classical infectious mononucleosis. These two cell lines exhibit distinguishing differences
in culture in parallel with reviously described differences in the cells when implanted into eonatal Syrian hamsters. These
observations suggest that the cell lines were derived from two pre-existing different classes or “stelines” of cells—one of
which resembles other cell lines derived from patients with acute lymphoblastic leukemia, whereas the other resembles cell
lines derived from other, patients with infecttious mononucleosis.
These studies were suported in part by research Grants C-6516 from the National Cancer Institute and FR-05526 from the Division
of Research Facilities and Resources, National Institutes of Health. 相似文献
19.
Ogle Jane W. Lange R. D. Dunn C. D. R. 《In vitro cellular & developmental biology. Plant》1978,14(11):945-950
Summary The in vitro production of the important regulatory of erythropoiesis, erythropoietin (Epo), is reviewed. It is concluded
that it is possible to produce almost routinely small quantities of Epo in tissue culture. Although such procedures offer
the potential to provide large quantities of the hormone for clinical use, the optimum culture conditions and mechanisms for
triggering Epo production have yet to be resolved.
This work was supported by Grants No. 74444 from The John A. Hartford Foundation, and HL 10567 from the National Institutes
of Health. 相似文献
20.
Root rot disease tolerant clones of turmeric variety Suguna of Curcuma longa L. were isolated using continuous in vitro selection technique against pure culture filtrate of Pythium graminicolum. Large amount of profuse, compact, creamish white callus was obtained from in vivo vegetative bud when cultured on LSBM fortified with 2,4-D (3 mg l−1) after 45 days of culture. Callus was challenged with pure culture filtrate of P. graminicolum to isolate viable callus within 30 days of culture, which was further subjected to pure culture filtrate treatment. After three cycles of treatment, four cell lines which are tolerant to culture filtrate was isolated through continuous in vitro selection and subcultured on regeneration medium LSBM fortified with BAP (4 mg l−1) along with the control non-selected callus to obtain complete plantlets through discontinuous in vitro selection technique. Plants regenerated from tolerant and non-selected calli were screened for disease tolerance by adopting in vitro sick plot technique. The data obtained from this experiment revealed a ratio of 225:49 tolerant: susceptible in vitro clones retrieved from tolerant callus. However, plants regenerated from the CL1a1 and non-selected calli were susceptible under in vitro sick plot technique. The root rot disease tolerant clones were hardened and established in soil with 90% survival frequency. 相似文献