共查询到20条相似文献,搜索用时 9 毫秒
1.
J.-C. Stoclet R. Andriantsitohaina N. L'heureux C. Martinez L. Germain F. Auger 《Cell biology and toxicology》1996,12(4-6):223-225
Relatively limited information is available regarding the mechanisms controlling vasomotricity in human vessels. Isolated vessels obtained from patients undergoing surgery were used to characterize the role of endothelial factors and to study coupling mechanisms between receptors, intracellular calcium, and contraction. However, these investigations are limited by the availability of tissues and many uncontrolled factors. Cultured human vascular cells were also used, were these cells rapidly lose at least some of their differentiated characters. Recently, a human blood vessel equivalent was constructed in vitro from cultured cells, using tissue engineering. This technique allowed us to obtain vessel equivalents containing intima, media, and adventitia layers or tubular media layer only. Contraction and rises in intracellular calcium produced by agonists were studied, indicating that such human vessel equivalents may provide valuable models for pharmacological studies. 相似文献
2.
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
3.
Summary Human arterial endothelial cells were cultured in vitro for up to 40 cumulative population doublings. Culture conditions similar
to those required for long-term propagation of human umbilical vein endothelial cells were employed. These included fibronectin-coated
culture vessels, 5 to 20% fetal bovine serum, endothelial cell growth factor, and heparin. Thoracic aorta endothelial cells
were larger than iliac artery endothelial cells. Both cell types stained positively for Factor VIII antigen by immunofluorescence.
A decrease in confluent density as a function of population doubling level was correlated with the appearance of large, senescent
cells in the cultures. Serum growth factors to which the arterial endothelial cells responded included insulin, transferrin,
epidermal growth factor, thrombin, and somatomedins. The effect of thrombin did not require the availabilty of the active
site of the protease. The effect of the somatomedins was only seen in the presence of heparin. Neither platelet-derived growth
factor nor hydrocortisone induced arteiral endothelial cell proliferation. These growth factor responses were also observed
on the part of human umbilical vein endothelial cells.
This work was supported in part by Public Health Service grants HL01030, HL01734, and AG00599. 相似文献
4.
Peroxidation products of polyunsaturated fatty acids may cause growth inhibition of cells in culture. This study was carried out to elucidate to what extent peroxidation products may be found in growth media, with and without cells and albumin, using thiobarbituric acid-reactive substances (TBARS) and protein carbonyl groups as measures of peroxidation. The growth of human microvascular endothelial cells was studied as influenced by docosahexaenoic (C22:6, n - 3), arachidonic acid (C20:4. n - 6), and serum albumin. Cell growth was strongly inhibited by the fatty acids, and the inhibition was related to the concentration of TBARS in the medium. Defatted albumin (0.5 g/100 ml) nullified the increase of TBARS in the medium and released the growth inhibition by the fatty acids. With polyunsaturated fatty acids (PUFA) there was a time- and concentration-dependent increase in media TBARS, observed both with and without cells, but the TBARS increase was somewhat greater in the presence of cells. Surprisingly, TBARS in cell-free media also increased somewhat upon increasing the albumin concentration from 0.5 to 5 g/100 ml, and the TBARS increase differed among various preparations of albumin. Unexpectedly, the albumin that had not been defatted gave the lowest TBARS values. The amount of protein carbonyl groups did not differ among various albumin preparations. It is concluded that PUFA may autooxidize in media used for cell cultures, and thereby cause an unspecific growth inhibition, which can be prevented by a low albumin concentration. However, even defatted albumin preparations may contain lipid peroxidation products, the causes and implications of which remain to be elucidated. 相似文献
5.
Roy J Kazi M Hedin U Thyberg J 《Differentiation; research in biological diversity》2001,67(1-2):50-58
Arterial smooth muscle cells grown in primary culture on a substrate of fibronectin in serum-free medium are converted from a contractile to a synthetic phenotype. This process is dependent on integrin signaling and includes a major structural reorganization with loss of myofilaments and formation of a large secretory apparatus. Functionally, the cells lose their contractility and become competent to migrate, secrete extracellular matrix components, and proliferate in response to growth factor stimulation. Here, it is demonstrated that the mitogen-activated protein kinases ERK1/2 play a vital role in the fibronectin-mediated modification of rat aortic smooth muscle cells. Immunoblotting showed that phosphorylated ERK1/2 (p44/p42) were expressed throughout the period when the change in phenotypic properties of the cells took place. Moreover, phosphorylated ERK1/2 accumulated in the nucleus as revealed by immunocytochemical staining. Additional support for an active role of ERK1/2 in the shift in smooth muscle phenotype was obtained by the finding that PD98059, an inhibitor of the upstream kinase MEK1, potently suppressed both the expression of phosphorylated ERK1/2 and the fine structural rebuilding of the cells. In conclusion, the observations point to an important and multifaceted role of ERK1/2 in the regulation of differentiated properties and growth of vascular smooth muscle cells. 相似文献
6.
J. Roy †¢ · M. Kazi† · U. Hedin† · J. Thyberg 《Differentiation; research in biological diversity》2001,67(1):50-58
Abstract Arterial smooth muscle cells grown in primary culture on a substrate of fibronectin in serum-free medium are converted from a contractile to a synthetic phenotype. This process is dependent on integrin signaling and includes a major structural reorganization with loss of myofilaments and formation of a large secretory apparatus. Functionally, the cells lose their contractility and become competent to migrate, secrete extracellular matrix components, and proliferate in response to growth factor stimulation. Here, it is demonstrated that the mitogen-activated protein kinases ERK1/2 play a vital role in the fibronectin-mediated modification of rat aortic smooth muscle cells. Immunoblotting showed that phosphorylated ERK1/2 (p44/p42) were expressed throughout the period when the change in phenotypic properties of the cells took place. Moreover, phosphorylated ERK1/2 accumulated in the nucleus as revealed by immunocytochemical staining. Additional support for an active role of ERK1/2 in the shift in smooth muscle phenotype was obtained by the finding that PD98059, an inhibitor of the upstream kinase MEK1, potently suppressed both the expression of phosphorylated ERK1/2 and the fine structural rebuilding of the cells. In conclusion, the observations point to an important and multifaceted role of ERK1/2 in the regulation of differentiated properties and growth of vascular smooth muscle cells. 相似文献
7.
Investigation into the interaction of methylparaben and erythromycin with human serum albumin using multispectroscopic methods 下载免费PDF全文
In this paper, the interaction of methylparaben and erythromycin with human serum albumin (HSA) was studied for the first time using spectroscopic methods including Fourier transform infrared (FTIR) spectroscopy and UV absorption spectroscopy in combination with fluorescence quenching under physiological conditions. The binding parameters were evaluated using a fluorescence quenching method. Based on Förster's theory of non‐radiation energy transfer, the binding average distance, r between the donor (HSA) and the acceptor (methylparaben and erythromycin) was evaluated. UV/vis absorption, FTIR, synchronous and 3D spectral results showed that the conformation of HSA was changed in the presence of methylparaben and erythromycin. The thermodynamic parameters were calculated according to the van't Hoff equation and are discussed. The effect of some biological metal ions and site probes on the binding of methylparaben and erythromycin to HSA were further examined. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献
8.
Long-term culture of human endothelial cells 总被引:9,自引:0,他引:9
Portia B. Gordon Ira I. Sussman Victor B. Hatcher 《In vitro cellular & developmental biology. Plant》1983,19(9):661-671
Summary Human umbilical vein endothelial cells can be grown in vitro for 28 passages (CPDL 58) in Medium 199 supplemented with newborn
bovine serum and a partially purified growth factor derived from bovine brain. Newborn bovine serum is superior to fetal bovine
serum for the proliferation of human umbilical vein endothelial cells seeded at low density in the presence of the growth
factor. The endothelial cells, which can be passaged every 7 to 10 d at a 1-to-5 split ratio, retain their morphological and
biochemical characteristics. The proliferation of cells seeded at low density (103/cm2) is proportional to the concentration of the growth factor present in the medium. The growth factor, which has an isoelectric
point between 5.0 and 5.5, can support cell proliferation at reduced serum concentrations; half-maximal growth is achieved
in medium containing the growth factor and 3% serum. The brain endothelial cell growth factor does not stimulate DNA synthesis
significantly in cultures of human skin fibroblasts.
This research was supported by grants from the U.S. Public Health Service (AG 01732, HL 16387, and HL 07080), the Cystic Fibrosis
Foundation, and the New York and American Heart Associations.
Victor B. Hatcher is an Established Fellow of the New York Heart Association and a recipient of the Ann Weinberg Cystic Fibrosis
Research Scholarship Award. 相似文献
9.
Linda J. Loretz Catherine A. Reznikoff 《In vitro cellular & developmental biology. Plant》1988,24(4):333-342
Summary We report the development of culture conditions which routinely support clonal growth of normal human uroepithelial cells
(HUC). Secondary cultures seeded at clonal densities and grown under conditions described herein have a colony-forming efficiency
(CFE) and colony size that will be useful for in vitro experiments. Primary cultures were dispersed to single cells and seeded
in a supplemented Ham's F12 medium containing 1% fetal bovine serum together with 3×105 lethally irradiated Swiss 3T3 feeder cells on plastic substrates preequilibrated with F12 medium containing 5 or 10% serum.
Using these conditions, the average CFE was 16.1±2.5%. A cloning efficiency of 4.9±1.5% was obtained under the same conditions
in serum-free F12+ when supplemented with a mixture of trace elements or 0.1 mM ethanolamine. The epithelial nature of the cloned cells was confirmed by morphology and by positive immunofluorescent staining
for human epithelial keratin proteins. To make this system useful for mutagenesis experiments, a clone of Swiss 3T3 feeder
cells resistant to 5 μg/ml 6-thioguanine (6TG) was derived from the parental cell line. This 6-TG-resistant Swiss 3T3 clone
supports HUC clonal growth with a CFE of 17.9±2.0% CFE. We also report clonal growth of HUC without feeder cells using supplemented
MCDB 170 medium containing 70 μg/ml bovine pituitary extract. The average cloning efficiency using these conditions was 5.7±1.7%.
This work was supported by NIH grant 29525 to C. A. R. L. J. L. is a recipient of National Science Foundation predoctoral
fellowship. 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.
Y Iwasaki J Iwasaki H C Freake 《Biochemical and biophysical research communications》1983,110(1):235-242
The human breast cancer cell line (T47D) has specific, high affinity calcitonin receptors and calcitonin-responsive adenylate cyclase. Human, salmon and [Asu1,7]eel calcitonin inhibited cell growth in a dose-related manner with almost equipotency. Analogues of human calcitonin demonstrated slight cell growth inhibition. We found extreme growth inhibition with daily treatment with dibutyryl cyclic AMP (10(-4) M). In contrast to calcitonin 1,25-(OH)2D3 had a biphasic effect on cell growth. Physiological doses (5 X 10(-10) M) of 1,25-(OH)2D3 stimulated growth of T47D, whereas treatment by supraphysiological amounts (2.5 X 10(-7) M) caused significant inhibition of growth. Calcitonin and 1,25-(OH)2D3 appeared to have additive effects. 相似文献
12.
Growth factor modulation of melanoma growth stimulatory activity mRNA expression in human malignant melanoma cells correlates with cell growth 总被引:2,自引:0,他引:2
This report demonstrates that the expression of melanoma growth stimulatory activity (MGSA) mRNA can be modulated in a positive fashion in the Hs294T human melanoma cell line by PDGF and MGSA. There is close correlation between MGSA expression and the pattern of cell growth in Hs294T cells. 相似文献
13.
Kristina Sundqvist Yun Liu Kristina Arvidson Kari Ormstad Lennart Nilsson Rune Toftgård Roland C. Grafström 《In vitro cellular & developmental biology. Animal》1991,27(7):562-568
Summary Human buccal epithelial cells have been reared from explants maintained in supplemented MCDB 153 medium. Primary epithelial
outgrowths show typical structural features and uniformly express keratins; subunit analyses demonstrate expression of keratins
5, 6, 14, 16/17, and 19. The cells exhibit up to 6% colony forming efficiency and divide at about 0.8 population doublings
per day on fibronectin/collagen-coated dishes at clonal density. Studies of markers of proliferation and differentiation in
buccal epithelial cells indicate that epidermal growth factor, cholera toxin, retinoic acid, and pituitary extract each exhibit
a distinctive ability to enhance growth and variably affect cell migration and cell surface area. Transforming growth factorβ-1 inhibits growth and increases surface area without affecting migration, involucrin expression, and cross-linked envelope
formation. Moreover, exposure of cells to fetal bovine serum, the tumor promoting agent 12-O-tetradecanoylphorbol-13-acetate or an elevated Ca2+ concentration (from 0.1 to 1 mM) inhibits growth and induces squamous differentiation as indicated by inhibition of migration, increases in surface area,
involucrin expression, or formation of cross-linked envelopes. The results show that epithelial cells can be reproducibly
derived from explant cultures of human buccal mucosa specimens and the cells transferred under serum-free conditions. Buccal
epithelial cells in culture undergo a pattern of growth and differentiation that mimics parakeratinization in vivo and variably
respond to several agents shown to modulate growth of cells that originate from other types of epithelia.
This work was supported in part by grants from the Swedish National Board of Laboratory Animals, the Swedish Medical Research
Council, the Swedish Natural Science Research Council, the Swedish Fund for Scientific Research Without Animal Experiments,
the Swedish Cancer Society, the Swedish Tobacco Company, and Lions Club International, Djurg?rden, Stockholm, Sweden. 相似文献
14.
Ichiro Ichinose Shuji Nakano Taito Esaki Tatsuhiko Koga Kenji Mitsugi Hidenori Yamada Yoshiyuki Niho 《In vitro cellular & developmental biology. Animal》1993,29(4):332-338
Summary A growth-inhibiting activity was identified in supernatants of the neoplastic V79 Chinese hamster cell line based on its ability
to inhibit the proliferation of the same cell line. The partially purified activity, provisionally termed “growth inhibiting
factor” (GIF) activity, inhibited the growth of a wide variety of human tumor cells, but not various normal human fibroblasts.
This species-nonspecific activity was reversible, saturable, and highly potent in tumorigenic cell lines, and was noted in
both monolayer culture and in soft agar. The inhibitory activity CIF was also exhibited in a chemically defined serum-free
medium supplemented with insulin and transferrin. GIF activity was stable to acid, heat, trypsin, and dithiothreitol but sensitive
to alpha-chymotrypsin. The pattern of growth modulation by GIF on V79 cells was apparently different from those exhibited
by bifunctional peptides such as transforming growth factor-beta, tumor necrosis factor-alpha, and interleukin-1-alpha. In
addition, GIF activity cannot be ascribed to these cytokines based on the physicochemical and immunologic properties. Although
GIF has yet to be purified to homogeneity, these data suggest that GIF might be a novel growth regulator which has a critical
role in regulating growth of V79 cells. The growth modulation of tumor cells by this tumor-derived growth inhibiting activity
suggested the presence of an autocrine growth regulatory mechanism even in tumor cells. 相似文献
15.
16.
Ge Gao Yang Zhang Yingjiu Chao Chaoshi Niu Xianming Fu Jianjun Wei 《Journal of cellular biochemistry》2019,120(12):19432-19441
Intracranial aneurysm (IA) is recognized as a lethal form of cerebrovascular disease mainly featured with a modulated phenotype of vascular smooth muscle cells (SMCs). It is generally believed that enhanced SMC proliferation and migration capabilities are the main characteristics in this process. In this study, we revealed that microRNA-4735 (miR-4735) participates in phenotypic modulation in a hypoxia-inducible factor-1 (HIF-1)-dependent manner of SMCs. miR-4735 targets the 3′-untranslated region of HIF-1. The downregulated expression of miR-4735 in IA tissues leads to elevated expression of HIF-1, which activates autophagy and promotes autophagy-mediated SMC proliferation and migration. Overexpression of miR-4735 suppressed HIF-1 expression and HIF-1-mediated autophagy, which led to impaired SMC proliferation and migration abilities. Forced expression of HIF-1 in miR-4735-overexpressed SMCs rescued the impaired SMC proliferation and migration abilities. In conclusion, miR-4735 plays an important role in phenotypic modulation in IA by regulating autophagy-promoted SMC proliferation and migration. 相似文献
17.
Many organisms occupy considerably different environments during individual's lifespan. We are interested in how the phenotypic
characteristics that are favourable in the earlier environment predict fitness in the later environment. High predictability
of fitness between the two consecutive environments suggests that the either the same traits are favored in both environments,
or that the favourable traits are genetically correlated. In this study, we ask how similarity of consecutive foraging environments
affects the phenotypic correlation of juvenile brown trout growth rate. More specifically, we used a genetically narrow stock
of hatchery-bred fish to contrast individual growth rates between high and low density hatchery environments, and thereafter
between hatchery and natural lake environment. As expected, growth rate was highly dependent on the environment, and the fish
showed considerable phenotypic plasticity. Furthermore, we found a strong positive correlation in growth rate between similar
foraging environments, for example, between high and low density hatchery stocks, and between hatchery and a lake with small
fish as main prey. However, hatchery growth did not predict growth rate in lakes where fish had to forage on bottom-dwelling
invertebrates. Our results suggest that when the consecutive environments differed dramatically with respect to traits that
fish use for foraging, relative performance of individual fish changed, earlier performance not being an accurate predictor
of performance in the new environment. In this case, fitness of the fish was determined by an environment-specific set of
traits that were not the same between the two consecutive environments. The result indicates that assessment of individual
performance may be highly environment specific in trout.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
18.
Monolayer cultures of human neuroblastoma cells were exposed to 915-MHz radiation, with or without sinusoidal amplitude modulation (80%) at 16 Hz, at specific absorption rates (SAR) for the culture medium and cells of 0.00, 0.01, 0.05, 0.075, 0.1, 0.5, 0.75, 1.0, 1.5, 2, or 5 mW/g. A significant increase in the efflux of calcium ions (45Ca2+) as compared to unexposed control cultures occurred at two SAR values: 0.05 and 1 mW/g. Increased efflux at 0.05 mW/g was dependent on the presence of amplitude modulation at 16 Hz but at the higher value it was not. These results indicate that human neuroblastoma cells are sensitive to extremely low levels of microwave radiation at certain narrow ranges of SAR. 相似文献
19.
Mitsuyama Takashi Hidaka Kouko Furuno Takashi Hara Nobuyuki 《Molecular and cellular biochemistry》1998,181(1-2):157-161
A 14-membered ring macrolide, erythromycin, acts not only as an antibacterial but also as an anti-inflammatory agent. We have previously reported that erythromycin modulates neutrophil functions and ameliorates neutrophil-induced endothelial cell damage through the action of cyclic AMP-dependent protein kinase (PKA) and nitric oxide (NO). We investigated the effect of erythromycin on human endothelial cell functions. Erythromycin enhanced intracellular calcium ion concentration ([Ca2+]i) of endothelial cells and NO release from endothelial cells. The enhancement of NO release from endothelial cells by erythromycin was abolished by addition of EGTA in the medium and was partially reduced by addition of H-89, an inhibitor of PKA. These results suggest that erythromycin enhances NO release from endothelial cells through the action of PKA and [Ca2+]i. In addition, constitutive NO synthase (cNOS) protein expression of endothelial cells was dose-dependently enhanced by treatment with erythromycin, which might also contribute to the enhancement of NO release from endothelial cells by erythromycin. The effect of erythromycin as an anti-inflammatory agent might be partially mediated through the enhancement of NO release from endothelial cells and the drug might be a useful tool for the investigation of cNOS of endothelial cells. 相似文献