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991.
Summary The ability of human erythroleukaemia K562 cells to take up aluminium from Al-transferrin and Al-citrate has been examined. Uptake from Al-transferrin was dose-dependent over the range 68–544 ng/ml of aluminium, and increased over a 12-day period. In contrast, uptake from Al-citrate was low even at an aluminium concentration of 6800 ng/ml and did not increase over time. Neither form of aluminium greatly affected cell growth. It is concluded that Al-transferrin, rather than Al-citrate, is the physiologically relevant form of this metal with respect to cellular uptake, but that any metabolic abnormalities induced by aluminium do not affect proliferation of this cell line. 相似文献
992.
造血细胞活力冷冻损伤的可恢复性 总被引:1,自引:0,他引:1
人骨髓冻存后其造血祖细胞活力有一定程度下降,本研究对这种下降的可逆性作了初步观察。结果发现,用双层法和单层法作CFU-GM培养时,未冻存骨髓集落产率相近,冻存骨髓双层法的CFU-GM产率高于单层法。骨髓细胞用20%FM-CM、PHA-LYCM、PHA-PMCM预孵育2h后,分别测定其CFU-GM、BFU-E与CFU-Mix,发现这种孵育过程对未冻存骨髓的集落产率无明显影响,而冻存骨髓的集落产率在孵育后可升高(GEMmeg除外)。说明骨髓造血祖细胞对冻存的损伤反应不均一,部分受损细胞在一定条件下可以恢复其增殖活力。这对于用冻存骨髓作骨髓移植可能有一定意义。 相似文献
993.
Armelle Baeza-Squiban Sylvie Romet Anne Moreau Francelyne Marano 《In vitro cellular & developmental biology. Animal》1991,27(6):453-460
Summary Primary cultures of rabbit tracheal cells were obtained as outgrowths from explants of tracheal mucosa. A 30% collagen substratum
containing serum and minimal essential medium was required for obtaining an outgrowth of epithelial cells keeping their differentiated
characteristics. The tracheal epithelial cells obtained near the explant in the first days of culture presented morphologic
similarities with normal tracheal epithelium. Cultures contained basal cells and epithelial polarized cells that exhibited
apical tight junctions and desmosomes. Ciliated cells stayed functional during all time culture. Their number slightly increased
at the beginning of the culture and then stayed constant when the total number of cells increased. Development of the outgrowth
was rapid and significant inasmuch as the outgrowth surface reached 30 times that of the explant after less than 8 days. This
was linked to cellular proliferation, as demonstrated by the incorporation of bromodeoxyuridine (BrdU) in phase-S nuclei and
the revelation of BrdU using an immunofluorescence technique. The epithelial nature of the outgrowth cells and the absence
of contamination with fibroblasts were established by positive staining with anti-keratin antibody and by negative staining
with anti-vimentin antibody, respectively.
This work was supported by DRET and by CIFRE grant awarded to S. R. 相似文献
994.
D. Needham 《Cell biochemistry and biophysics》1991,18(2):99-121
Studies that examine the shear- and abrasion-sensitivity of proliferating cells are important in order to understand the behavior
of hybridoma cells in bioreactor culture and metastasizing cancer cells in the bloodstream. Little is known about the link
between morphology, structure, and mechanical properties of a given cell line, especially with respect to variations throughout
the cell cycle. In our experiments with GAP A3 hybridoma cells, distinct cell morphologies were identified and correlated
with phases of the cell cycle by video microscopic observation of synchronized cells, and of individual cells that were followed
throughout their cell cycle. Micropipet manipulation was used to measure the geometrical (cell volume) and mechanical (apparent
cell viscosity) properties of single cells. As the cell cycle progressed at 37°C, an increase in cell volume from 1400 μm3 to 5700 μm3 was accompanied by an increase in apparent cell viscosity from 430 poise to 12,000 poise, consistent with an accumulation
of more cytoplasmic material in the “older” cells. Hybridomas are representative of the various leukemias derived from hemopoietic
cells, and even though as a whole, they appeared to be rather shear-insensitive, the wide range of property values demonstrates
that a given cell line cannot be characterized by a single value for any one property, and that properties must be related
to the cell cycle when considering proliferating cells. It is interesting to see if distinct stages in the metastatic sequence
of events might correlate with any of these physical features of the cell cycle, irrespective of cell type or cell line. For
example, the cytokinetic doublet could represent a fragile structure that may fail and produce cell death under fluid-shear
conditions that would not affect the cells at any other stage in the cell cycle. Identifying such cell cycle-dependent features
in metastasizing cancer cells could lead to a better understanding of the metastatic process and to possible clinical treatments
directed at making cells more shear- and abrasion-sensitive, and therefore, more likely to be killed by the natural hydrodynamic
forces of the circulatory system. 相似文献
995.
Hiroyoshi Hoshi Yuji Takagi Keizo Kobayashi Masakazu Onodera Taneaki Oikawa 《In vitro cellular & developmental biology. Animal》1991,27(7):578-584
Summary We have developed an improved serum-free medium to optimize the cell growth of bovine granulosa cells. The cells on collagen-coated
culture plates proliferated extensively in a nutrient medium supplemented with insulin, heparin binding growth factor-2 (HBGF-2),
lipoprotein, and bovine serum albumin (BSA). The cell doubling time at logarithmic phase and final cell density at confluent
cultures were equal to those of cultures grown in the presence of medium supplemented with optimal concentration (10%) of
fetal bovine serum (FBS). Whereas HBGF-2 or insulin alone had a small mitogenic effect of granulosa cells, lipoprotein or
BSA did not. When lipoprotein, BSA, or insulin was added together with HBGF-2, synergistic cell proliferation was observed
in all combinations. Insulin or lipoprotein had an additive mitogenic stimulation of these cells in the presence of BSA. After
granulosa cells were subcultivated in a serum-containing medium until three generations [8.5 cumulative population doubling
level (CPDL)], subsequent subcultivation of the cells in a complete serum-free medium could be achieved up to six generations
(14.4 CPDL). These results demonstrate that this serum-free medium can support the optimal cell growth and long-term subcultivation
of bovine granulosa cells. 相似文献
996.
Javier Turnay Nieves Olmo José G. Gavilanes María A. Lizarbe 《In vitro cellular & developmental biology. Animal》1991,27(6):447-452
Summary Fibroblastlike primary cells have been obtained from human colon adenocarcinoma explants. Such cells disappear during cell
culture and thus have not been previously studied. These cells have a number of altered phenotypic characteristics: a) morphology;
b) growth behavior and adherence to culture substrate (they required 3 h for 90% attachment and only presented a flattened
morphology 40 h after platting); and c) collagen metabolism. Increased protein biosynthesis (about double than control colon-derived
fibroblasts) and maintained ability for collagen biosynthesis have been observed for the tumor-associated fibroblastlike cells.
Thus, the collagen to noncollagenous proteins ratio was decreased for these cells. They exhibited an altered type I:type III
collagen (5:1 instead of 3:1 in colon fibroblasts) and procollagen (2:1 against 5:1 in colon fibroblasts) ratios as well as
a decreased secretion of collagen with an abnormal deposition of procollagens in the cell layer. These studies show a permanent
phenotypic alteration in the tumor-associated fibroblastlike cells. 相似文献
997.
We have previously shown that human recombinant methionyl manganese superoxide dismutase (MnSOD) is more efficient than CuZnSOD as an anti-inflammatory agent in a model of acute inflammation (Carrageenan-induced pau edema). This effect was attributed to the prolonged half-life of MnSOD in blood (t1/2 = 6 h vs. 10 min. respectively). In the present study, the two enzymes were compared in terms of their effectiveness in two systems: (I) Adjuvant-induced arthritis in rats, which is considered to be a model for the chronic situation of rheumatoid arthritis and (2) Bleomycin-induced lung fibrosis. which is a chronic situation believed to be mediated by oxygen free radicals.
Rats inflicted with adjuvant arthritis were treated during the period of maximal joint swelling (Days 15-21 after adjuvant injection) with MnSOD or CuZnSOD (12 to 50mg/kg, i.p. daily). MnSOD administration resulted in a 50-75% reduction of paw swelling, as well as inhibition of the elevation of serum globulins. A similar treatment with CuZnSOD gave merely marginal effects.
In the second system, lung fibrosis was induced in rats by intratracheal administration of bleomycin. MnSOD (50mg/kg, s.c.), administered 2 h before and then 2 and 4 days after bleomycin, markedly inhibited lung fibrosis, as evident from lung weight and collagen content measured by the 3rd week. By contrast, CuZnSOD administration did not give a significant effect. The results indicate that MnSOD is superior to CuZnSOD in the treatment of chronic inflammatory processes. In addition, they lend further support to the involvement of oxygen free radicals in bleomycin toxicity. 相似文献
Rats inflicted with adjuvant arthritis were treated during the period of maximal joint swelling (Days 15-21 after adjuvant injection) with MnSOD or CuZnSOD (12 to 50mg/kg, i.p. daily). MnSOD administration resulted in a 50-75% reduction of paw swelling, as well as inhibition of the elevation of serum globulins. A similar treatment with CuZnSOD gave merely marginal effects.
In the second system, lung fibrosis was induced in rats by intratracheal administration of bleomycin. MnSOD (50mg/kg, s.c.), administered 2 h before and then 2 and 4 days after bleomycin, markedly inhibited lung fibrosis, as evident from lung weight and collagen content measured by the 3rd week. By contrast, CuZnSOD administration did not give a significant effect. The results indicate that MnSOD is superior to CuZnSOD in the treatment of chronic inflammatory processes. In addition, they lend further support to the involvement of oxygen free radicals in bleomycin toxicity. 相似文献
998.
Summary A serum-free culture system supplemented with neural tissue extract for normal and tumor human esophagi was applied to the
culture of mouse esophageal epithelium. Similar to mouse mesenchyme and skin epithelium, esophageal epithelial lines (MEE)
emerged after serial culture. The cells had an apparent unlimited life span but retained morphology and other characteristics
of normal epithelial cells. The cells formed a small cyst consisting of keratined squamous epithelium in syngenic hosts. A
screen for growth factors that stimulated growth of the nonmalignant MEE cells in the absence of neural extract revealed that
epidermal growth factor (EGF) and heparin-binding (fibroblast) growth factors (HBGF) were most effective. An HBGF-like activity
was apparent in extracts of rapidly proliferating but not quiescent MEE cells at low or confluent densities. A cloned cell
line (MEE/C8) was selected from MEE cell cultures in the absence of neural extract. MEE/C8 cells proliferated independent
of either EGF or HBGF at rates equal to MEE cells, cell extracts exhibited HBGF-like activity at all stages of proliferation,
and the cells formed large invasive tumors in syngenic hosts. The HBGF-like activity present in extracts of tumorigenic MEE/C8
and proliferating nonmalignant MEE cells had properties similar to HBGF-1 (acidic fibroblast growth factor). These results
constitute a cultured mouse esophageal epithelial cell model for study of conversion of immortalized premalignant cells to
malignant cells, and suggest that conversion from a state of cell cycle-dependent autocrine expression of one or more members
of the HBGF family to a state of constitutive expression correlates with and may contribute to malignancy.
The work was supported in part by grants CA37589 and DK35310 to Dr. McKeehan, from the National Cancer Institute, Bethesda,
MD. 相似文献
999.
Maryceline T. Espanol Lawrence Litt Guo-yuan Yang Lee-Hong Chang Pak H. Chan Thomas L. James Philip R. Weinstein 《Journal of neurochemistry》1992,59(5):1820-1828
Metabolic tolerance of low intracellular pH (pH(i)) was studied in well-oxygenated, perfused, neonatal, rat cerebrocortical brain slices (350 microns thick) by inducing severe hypercapnia. In each of 17 separate experiments 80 brain slices (approximately 3.2 g wet weight) were suspended in an NMR tube, perfused with artificial CSF (ACSF), and studied at 4.7 T with 31P and 1H NMR spectroscopy. Spectra obtained every 5 min monitored relative concentrations of lactate or high-energy phosphate metabolites, from which pH(i) and extracellular pH were determined. Unperturbed slice preparations were metabolically stable for > 10 h, with no significant changes occurring in pHi, ATP, phosphocreatine (PCr), inorganic phosphate, or lactate. Different levels of hypercapnia were produced by sequentially perfusing slices with the following different ACSF batches, each having previously been equilibrated with a specific mixture of CO2 in oxygen: (a) 10% CO2, 15 min of perfusion; (b) 30% CO2, 15 min of perfusion; (c) 50% CO2, 15 min of perfusion; (d) 70% CO2, 30 min of perfusion; (e) 50% CO2, 15 min of perfusion; (f) 30% CO2, 15 min of perfusion; and (g) 10% CO2, 15 min of perfusion. At the completion of this protocol slices were again perfused with fresh ACSF that was equilibrated with a 95% O2/5% CO2 gas mixture. In each of five separate 1H and 31P experiments, brain slices were recovered within 2 h after termination of exposure to high CO2. The pHi was determined from measurements of the chemical shift difference between phosphoethanolamine and PCr, using a calibration curve obtained for our preparation.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
1000.
Specific Expression of N-Acetylaspartate in Neurons, Oligodendrocyte-Type-2 Astrocyte Progenitors, and Immature Oligodendrocytes In Vitro 总被引:14,自引:0,他引:14
Jutta Urenjak Stephen R. Williams David G. Gadian Mark Noble 《Journal of neurochemistry》1992,59(1):55-61
To test the specificity of N-acetylaspartate (NAA) as a neuronal marker for proton nuclear magnetic resonance (1H NMR) spectroscopy, purified and characterized cultured cells were analyzed for their NAA content using both 1H NMR and HPLC. Cell types studied included cerebellar granule neurons, type-1 astrocytes, meningeal cells, oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells, and oligodendrocytes. A high concentration of NAA was found in extracts of cerebellar granule neurons (approximately 12 nmol/mg of protein), whereas NAA remained undetectable in purified type-1 astrocytes, meningeal cells, and mature oligodendrocytes. However, twice the neuronal level of NAA was found in O-2A progenitors grown in vitro. In addition significant levels of NAA were also detected in cultures of immature oligodendrocytes. Our data partly support previous suggestions that NAA may be a useful neuronal marker for 1H NMR spectroscopic examination of the adult brain. However, they also raise the further possibility that alterations of NAA associated with some specific brain disorders, particularly disorders seen in newborn and young children, may reflect abnormalities in the development of oligodendroglia or their precursors. 相似文献