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1.
Summary Adult human prostatic epithelium was cultured in a defined medium consisting of RPMI 1640 supplemented with transferrin, insulin, epidermal growth factor, dexamethasone, and vitamin A. In the presence of insulin, stabilized with zinc, maximum epithelial multiplication was obtained at an insulin concentration of 0.03 to 0.1 U/ml, corresponding to a zinc concentration of 1.4×10−7 M. At higher insulin concentrations, growth stimulation declined. Zinc-free insulin, on the other hand, stimulated cell multiplication with an optimum concentration of 0.3 to 1.0 U/ml. At this concentration the maximum growth was twice that obtained with zinc-stabilized insulin. Results demonstrate that growth inhibition caused by zinc limits the concentration of zinc-stabilized insulin, which can be used in serum-free, defined culture media. This work was supported by the Division of Cancer Cause and Prevention, National Cancer Institute, DHHS Grant No. CA-28279 to Webber.  相似文献   

2.
The effect of recombinant murine interferon-gamma on the growth of Chlamydia trachomatis was analyzed in a mouse fibroblast cell line (McCoy cells). Murine interferon-gamma had a very potent anti-chlamydial activity, although minimally affecting cellular proliferation. Over 95% inhibition of chlamydial inclusions was obtained at a concentration of 1 U/ml of interferon. At a concentration of 1 U/ml of murine interferon-gamma, there was minimal inhibition of the proliferative capacity of McCoy cells. Approximately 50% inhibition of cell growth was obtained with a concentration of 10 U/ml of interferon. Varying concentrations of tryptophan in the medium did not alter either the anti-chlamydial or the anti-proliferative activity of the interferon.  相似文献   

3.
Growth factors may play an important role in regulating the growth of the proximal tubule epithelium. To determine which growth factors could be involved, we have investigated the mitogenicity of various purified factors in rat kidney proximal tubule epithelial (RPTE) cells cultured in defined medium. Fibroblast growth factors, aFGF (acidic FGF) and bFGF (basic FGF), stimulate DNA synthesis in a dose-dependent manner, with ED50 values of 4.5 and 3.2 ng/ml, respectively; their effects are not additive. With cholera toxin in the medium, both aFGF and bFGF can replace insulin or epidermal growth factor (EGF) to attain the maximum level of cell growth, but they cannot replace cholera toxin. Cholera toxin specifically potentiates the effects of FGFs on DNA synthesis. At high cell density, both insulin and insulin-like growth factor 1 (IGF-1) induce DNA synthesis more effectively than EGF, FGFs and cholera toxin. The high concentration (0.2-1.0 microgram/ml) of insulin required for cell growth can be replaced by a low concentration of IGF-1 (10-20 ng/ml), indicating that insulin probably acts through a low affinity interaction with the IGF-1 receptor. Transforming growth factor-beta 1 (TGF-beta 1) inhibits DNA synthesis induced by individual factors and combinations of factors in a concentration-dependent manner. Northern blot analysis shows that mRNA for TGF-beta 1, IGF-1, and aFGF, but not bFGF are present in rat kidney. Western blot analysis and bioassay data confirmed that the majority of FGF-like protein in rat kidney is aFGF. The data suggest that in addition to EGF, IGFs, and TGF-beta, FGFs may also be important kidney-derived regulators of proximal tubule epithelial cell growth in vivo and in vitro.  相似文献   

4.
Effects of insulin on glucose metabolism in isolated human fat cells   总被引:3,自引:0,他引:3  
Isolated fat cells were used for the study of in vitro effects of insulin on glucose metabolism in human and rat adipose tissue. In human subcutaneous fat cells, effects of insulin could be detected at concentrations of glucose in the medium from 1 to 10 micro moles/ml. Cellular responsiveness was inversely proportional to the glucose level. At a constant concentration of 6 micro moles of glucose per ml, the effects of insulin at various concentrations up to 500 micro U/ml were investigated. At the highest concentration, which gave the maximal response, there was a 100% increase in the conversion of glucose-U-(14)C to glyceride-glycerol and a 40% increase in glucose oxidation. The dose-response curve was steepest between 2 and 20 micro U/ml. Rat epididymal fat cells were much more responsive to insulin. Glucose lipogenesis and pentose cycle activity could also be demonstrated in rat cells, whereas these activities could not be shown in fat cells from human omental and subcutaneous tissue. The findings for human cells are attributed to changes in cellular activity during preparation.  相似文献   

5.
The influence of gastric inhibitory polypeptide (GIP) on fatty acid incorporation into adipose tissue (FIAT) was studied in the rat on epididymal fat pads at concentrations amounting to 1, 2 and 4 ng/ml. Without insulin in the incubation medium, GIP induced a slight though significant FIAT decrease with a maximum of 9% for 2 ng/ml concentration. In the presence of rat insulin (100 μU/ml), it significantly enhanced the insulin-induced FIAT increase, that progressed from 106.4% of the basal value to 110.5% for 1 ng/ml concentration (P < 0.025) and to 118.2% for 4 ng/ml concentration (P < 0.0025).The existence of such a phenomenon as well as that of an hyperactive enteroinsular axis in obese subjects could represent two important factors in the development of obesity.  相似文献   

6.
In order to develop a defined cultivation medium for HL-60 cells, we cultivated these cells in a serum-free suspension medium and tested the effect of various growth factors. Of the factors tested, granulocyte/macrophage colony-stimulating factor was most active in growth stimulation. A much lower effect was obtained with granulocyte colony-stimulating factor and transferrin. No effect was found with interleukin-3 and insulin. Granulocyte colony-stimulating factor was the only growth factor tested that also induced differentiation as judged by the nitroblue tetrazolium test. Growth of HL-60 cells in medium containing granulocyte/macrophage colony-stimulating factor (125 U/ml) and transferrin (5 micrograms/ml) as the only protein factors was similar to growth in medium containing 10% serum. No increase in spontaneous differentiation of HL-60 cells in this defined medium was observed. Physiological concentrations of retinol bound to retinol-binding protein and retinyl ester in chylomicron remnants reduced proliferation as well as the level of c-myc oncoprotein and induced differentiation of HL-60 cells cultivated in defined medium. Hence, this defined medium may be useful when studying the function of retinoids in HL-60 cells.  相似文献   

7.
The effects of 17 beta-estradiol (E2), epidermal growth factor (EGF) and insulin, alone or in association on guinea-pig uterine epithelial cell proliferation were examined in serum-free culture conditions. Primary cultures of epithelial cells were made quiescent by serum depletion, then incubated in a chemically defined medium. In this medium, insulin increased DNA synthesis but not in a dose-dependent manner for concentrations ranging from 0.2 to 10 micrograms/ml. A significant effect of EGF was found only for the highest concentration tested (100 ng/ml). E2 alone or in the presence of insulin (1 microgram/ml) had no effect whatsoever on the concentration tested (10(-10)-10(-5)M). Insulin (10 micrograms/ml) plus EGF (100 ng/ml) exerted on DNA synthesis and cell proliferation a significant additive effect which was identical to the growth stimulation induced by 10% fetal calf serum. The effects of insulin plus EGF were not modified by the addition of E2. These findings suggest that E2 is not directly mitogenic for uterine epithelial cells in defined culture conditions and that the mitogenic response to optimal concentration of insulin plus EGF is independent of E2.  相似文献   

8.
方茂楠  苏晓荣  卫静  谢利芳  刘涛 《生物磁学》2011,(19):3706-3708
目的:探讨不同浓度外源性胰岛素在不同浓度葡萄糖情况下对β TC-3细胞胰岛素分泌的影响。方法:取对数生长期的13TC3细胞分三组,即低糖组、中糖组、高糖组(葡萄糖浓度分别取1.0mmol/L、3.Ommoi/L、20.Ommol/L)。每组分0、5、10、15、100、500、5000和50000μU/ml胰岛素八个亚组(其中0μU/ml作为对照组)。刺激10分钟后取上清液测C肽。结果:在高糖组中,C肽分泌量无明显差异;在中糖组中,10μU/ml和15μU/ml两组相对对照组C肽分泌量显著增加,50000μU/ml组C肽分泌量则相对对照组出现减少,其余3个亚组无明显改变;在低糖组中,c肽分泌量除5000μU/ml组减少外。其它亚组C肽分泌量无明显差畀。结论:胞外胰岛素在适宜葡萄糖浓度时,对BTC3细胞胰岛素分泌的反馈影响呈剂量依赖关系。  相似文献   

9.
采用3H-TdR参入法,测定碱性成纤维细胞生长因子(bFGF)、胰岛素和内皮素-1(ET-1)对体外培养的大鼠肾小球系膜细胞(MC)增殖的影响,以及胰岛素与bFGF或ET-1促MC增殖的协同作用。结果表明,不同浓度的bFGF(5-200ng/ml)和胰岛素(0.1-2.4U/ml)均显著升高MC的3H-TdR参入值(cpm值)(P<0.01)。ET-1对MC的cpm值的影响依剂量不同呈现两种不同的效应,在10-9-10-7mol/L时,随着浓度的升高,MC的cpm值明显升高(P<0.01),并以10-8mol/L作用最强;当升高到10-6mol/L时,MC的cpm值出现降低趋势。胰岛素与bFGF或低浓度ET-1(≤10-8mol/L)共同作用于MC时,MC的cpm值明显高于二者单独作用之和(P<0.01),与高浓度ET-1(>10-7mol/L)共同作用于MC时,MC的cpm值小于二者单独作用之和(P>0.05)。上述结果说明,胰岛素、bFGF和ET-1均能显著促进MC增殖;胰岛素与bFGF或低浓度的ET-1促MC增殖具有正协同作用,与高浓度ET-1呈现负协同作用。  相似文献   

10.
Maximum growth for Rhizopus sp. A-11 was obtained at a zinc ion concentration of 0.7 ppm in a liquid medium. Glucoamylase (GA, EC 3.2.1.3) production in Rhizopus sp. A-11 was maximized at 710 U/ml, at the presence of 75 ppm for calcium and 0.7 ppm of zinc ions in liquid medium. Zinc ion is known as an essential biometal for Rhizopus growth; however, growth was inhibited by the zinc ion concentration, not maximized. Although calcium ion was not necessary to Rhizopus growth, GA production using Rhizopus sp. A-11 was markedly stimulated by calcium ion concentration over 75 ppm in the liquid medium. The GA productivity of the present liquid culture was about 4.4 times higher than that of the solid state culture, based on the unit starch amount in the liquid and solid media carbon source. The characteristics of the GA produced by the Rhizopus sp. A-11 liquid culture were interesting; that is, almost all the GA produced was classified as raw starch-digesting GA (GA-I). Secreted protein in the culture liquid after 30 h was nearly GA, and had a limited amount of impure protein. As a result, it was found that using a Rhizopus culture in a specified metal-ion regulated medium was an effective method for producing GA. Thus the present culture method was renamed the "metal-ion-regulated liquid culture method".  相似文献   

11.
High yields (1939 U/ml) of an alkaline protease were obtained in batch fermentation of a Bacillus sp. using a response surface methodology. The interaction of four variables, viz., starch, peptone, incubation time, and inoculum density, suggested inoculum density to be an insignificant variable. However, incubation time had a profound effect on protease yields at all the concentrations of carbon and nitrogen used. The response surface raised and flattened with increase in time of incubation, and maximum protease production up to 1939 U/ml was obtained after 96 h of incubation. The model equation obtained was validated experimentally at maximum starch (15 mg/ml) and peptone (7.5 mg/ml) concentration with increased incubation time up to 144 h in the presence of minimum inoculum density (1%). An overall 2.6-fold increase in protease production was obtained as compared with mean observed response (750 U/ml) at zero level of all variables. Received: 19 July 2001 / Accepted: 15 August 2001  相似文献   

12.
Hemorrhagic shock was produced by bleeding conscious rats to a mean arterial pressure of 40 mm Hg, which was maintained for 2 h. Basal glucose uptake by isolated soleus muscle from normal rats and rats subjected to hemorrhagic shock ('shock' muscles) increased with the increase ib medium glucose concentration. Uptake values were similar in both groups of muscles. This indicates that there were no alterations in the basal glucose carrier mechanism during shock. Whereas insulin (0.1 U/ml) stimulated glucose uptake in control muscles under aerobic as well as under anaerobic conditions, it had no stimulatory effect in 'shock' muscles under either environment. Maximal stimulation of glucose uptake in 'shock' muscles was observed at an insulin concentration of 0.2 U/ml. The ability of muscle to bind insulin was not altered during shock. The present experiments indicate that insulin responsiveness to tissues is altered in shock. This could be due to alterations in the insulin sensitivity of the glucose carrier mechanism during shock.  相似文献   

13.
Mitogenic properties of the insulin derived from pig brain were compared with the action of pancreatic (standard) pig insulin and epidermal growth factor (EGF) using the culture of Swiss 3T3 cells. The brain insulin, likely as the pancreatic insulin, induced uptake of 14C-thymidine by resting cells in a dose-dependent manner at concentration of 0.5-2.0 micrograms/ml in culture medium. However, at equal concentrations of these hormones the proliferating effect of the brain insulin appeared to be 2-fold higher than the effect of the pancreatic hormone. At the same time the mitogenic action of both hormones was lower than that of EGF (10 ng/ml). The additive effect of the brain insulin and EGF, administered in combination, was more pronounced than the effect of the pancreatic insulin combined with EGF. The data obtained suggest a possible participation of brain insulin in the process of nerve cell proliferation.  相似文献   

14.
SYNOPSIS. In chemically defined media at carbohydrate concentrations ≧ 0.5% (w/v) Tetrahymena pyriformis W multiplied more rapidly, developed larger cells, and achieved greater growth as measured by optical density when carbohydrate was provided as dextrin rather than glucose. In media containing 0.3 mg/ml of amino acid nitrogen, growth increased with glucose concentration from 0.1 to 1%, did not change significantly to 3%, and was sharply inhibited at higher glucose levels. With dextrin, maximum growth paralleled carbohydrate concentration from 0.1 to 3%. At higher N levels the inhibitory concentration of glucose was lowered, but growth in dextrin media was not affected except at N concentrations that were inhibitory independent of carbohydrate source. At 1% carbohydrate levels, total cell protein per ml of culture was 60% greater, protein per cell approximately 50% greater, and cells were 1.5 to 2 times larger in media with dextrin than with glucose. Comparable differences in protein synthesis were observed at 2% carbohydrate levels and efficiency of conversion of substrate-N to protein-N was greater in the medium with dextrin than glucose.
Growth as measured by optical density in media with 0.3 mg/ml of N and 1 or 2% (w/v) of dextrin was not significantly reduced by the simultaneous presence of 1 or 2% glucose. This observation appeared to negate osmotic pressure as an explanation of reduced growth in the presence of glucose. At higher osmolar concentrations osmotic pressure appeared to be a major determinant of overall growth but not of cell size.  相似文献   

15.
A serum-free defined culture system has been developed that maintains follicle-stimulating hormone (FSH)-dependent differentiation of porcine granulosa cells from small follicles for up to six days in culture. Confluent monolayers of epithelioid cells were established after culture on fibronectin-coated culture dishes (FBN, 2 micrograms/cm2) in nutrient medium supplemented with human low-density lipoprotein (LDL, 10 micrograms/ml), insulin (I, 1 microgram/ml), and thrombin (TH, 1 NIH U/ml). Each of these factors was necessary to maintain the epithelioid morphology of the monolayers that attained 70% of the protein content and 71% of the cell number of replicate cultures maintained in nutrient medium supplemented with 10% fetal calf serum and insulin. Addition of FSH to the FBN/LDL/I/TH-supplemented cultures resulted in dose-dependent increases in progesterone secretion and [125I]-iodo-human chorionic gonadotropin (hCG) binding comparable to those obtained in the cultures containing serum. These results indicate that the attachment, epithelioid morphology, and differentiated function of porcine granulosa cells (GCs) can be maintained in defined culture conditions. This culture system will facilitate study of the effects of growth promoters and differentiative agents on GC function in the absence of poorly defined serum supplements.  相似文献   

16.
Zinc inhibited the colony formation of Cloudman S-91 murine melanoma cells in a dose dependent manner with an ID50 of 3.4 ug/ml. Total inhibition of the melanoma colony-forming units occurred at a zinc concentration of 4.42 ug/ml. In the presence of dexamethasone the ID50 for zinc inhibition was reduced by 49% and total inhibition of anchorage-independent growth occurred at the achievable in vivo zinc concentration of 3.0 ug/ml. Dexamethasone and zinc in combination effected a greater than additive inhibition of the murine melanoma colony-forming units. Statistical evaluation of these results showed that zinc and dexamethasone interacted synergistically to inhibit the formation of murine melanoma colonies.  相似文献   

17.
Effects of insulin on cultured hog thyroid cells were examined in a chemically defined medium supplemented with thyrotropin. Thyrotropin-induced follicle reconstruction and recovery of functional activities were stimulated by the addition of insulin to the culture medium. The iodide uptake and organification at the maximum stimulation exceeded 3 times those of the control values. The minimum effective dose of insulin was 60 microU/ml. The insulin stimulation occurred after a latency period of 24 h. Since no growth promoting effect of insulin was observed during the cultivation period in the present study, insulin appears to act in concert with thyrotropin in regenerating the characteristic properties of the differentiated cells during cultivation.  相似文献   

18.
Cultivation of Aureobasidium pullulans in medium with a low concentration of yeast extract (0.4 g/l) led to a decrease in the growth rate early in the fermentation as compared to cultivations in medium with high concentration of yeast extract. When this medium was supplemented with zinc and iron the cultivation closely resembled that obtained in medium with high concentration of yeast extract (4.0 g/l). The culture retained a high growth rate throughout the fermentation and the initiation of the mycelial to yeast (M-Y) transition and the exopolysaccharide production was delayed. In a defined medium or in defined medium without iron only a little exopolysaccharide was produced and the yeast fraction of the total biomass at the onset of the stationary phase was 22%–25%. However, cultivation in the defined medium without zinc resulted in a high production of exopolysaccharide and an increased intensity of the M-Y transition, which led to a yeast fraction of 41%.  相似文献   

19.
125I-Insulin binding to isolated brain microvessels from control, streptozotocin diabetic, and insulin-treated diabetic rats was measured. The binding was highest in the control (21.1 +/- 1.8%/mg capillary protein) and lowest in the diabetic (14.8 +/- 1.9%, p less than 0.01) animals. Administration of 2 U of protamine zinc insulin per day increased the maximum binding in the diabetic rats to 17.2 +/- 2.1%. Scatchard analyses of the binding showed that the major difference between the diabetic and the control animals was a decrease in the number of both high- and low-affinity sites in the diabetic animals. To test whether the failure of up-regulation in the hypoinsulinemic diabetic animal was related to an inherent defect in the endothelial cell or resulted from the diabetic milieu, cultured brain endothelial cells were tested for their capacity to up- and down-regulate their insulin receptors in vitro. In response to 100 ng/ml insulin for 12 h, these cells down-regulated their insulin receptors. When the insulin was removed, the insulin receptors returned to control levels. These studies showed that in vitro brain capillary endothelial cells have the capacity to increase their insulin receptors in response to a low-insulin environment, whereas in vivo the microvessels decrease their insulin receptors in response to diabetes.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

20.
Summary Biosynthetic human epidermal growth factor (Bh-EGF) induced dose-dependent synthesis and secretion of neutral mucin glycoprotein when the fundal cells isolated from rabbit stomach were cultured in serum-free medium containing Bh-EGF at concentrations as high as 10 to 100 ng/ml. At these high concentrations, Bh-EGF had no effect on the cell growth. In marked contrast, much lower concentrations from 0.1 to 1.0 ng/ml of Bh-EGF failed to stimulate mucin synthesis, but enhanced proliferation of the cells. Electrophoretic pattern of the mucin secreted from the cultured mucosal cells was very similar to that of the authentic mucin obtained from rabbit stomach. Maximal secretion of the mucin from the cells was observed at Hour 96 of the culture. Although fetal bovine serum (5%) and insulin (0.5 μg/ml) also stimulated the mucosal cells, both in growth and in mucin synthesis and release, the enhancing activity of the mucin synthesized and released by Bh-EGF at a concentration of 100 ng/ml per microgram DNA of cultured cells was far superior to that of 5% fetal bovine serum and 0.5 μg/ml insulin.  相似文献   

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