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31.
本文报告一种为适于杂交瘤细胞生长“无血清”培养基-适量的成牛血清低分子成分超滤液、10mg/L转铁蛋白(T)、10mg/L胰岛素(I)、20μmol/L乙醇胺(E)、40nmol/L硒酸钠(S)等诸补充成分替代胎牛血清加到基础液中-也完全适用于培养肿瘤细胞,且观察到与之相似的规律性结果,癌细胞在本“无血清”培养若的生长水平达到在含胎牛血清培养基中生长的水平。对于癌细胞的生长,LMW-CS与T、I、 相似文献
32.
In areas with short growing seasons, poor early vegetative growth of soybean (Glycine max [L.] Merr.) is often attributed to the restrictive effect of cool soil conditions on nodulation and N2-fixation by this subtropical grain legume. However, there are few studies regarding potential genetic variability of soybean and Bradyrhizobium japonicum genotypes for nodulation at cool root-zone temperatures (RZT). Experiments were conducted to (1) test for a threshold temperature for low RZT inhibition of soybean nodulation and (2) ascertain whether this threshold temperature response depends mainly on the micro- or macrosymbiont. In experiment 1 soybean seedlings (Glycine max [L.] Merr. cv. Maple Arrow) were inoculated with 1 ml of a log phase culture of B. japonicum strain 532C, H8 or H15 (the latter two strains were isolated from cold soils of Hokkaido, northern Japan) and maintained at either 16, 17.5, 19 or 25°C RZT. In experiment 2 seedlings of cv. Maple Arrow and a cold-tolerant Evans isoline were combined with strain 532C and two Hokkaido strains (H5, H30) at both 19 and 25°C RZT. Results indicated that N2-fixation at 44 days after inoculation was substantially reduced (30–40%) by RZT as high as 19°C, due to development of less nodule mass and to a delay in the onset of N2-fixation and a small decrease in the number of nodules formed. However, the number of nodules formed was sharply reduced and the time required for the first appearance of nodules was significantly delayed below an RZT of 17.5°C. Differences between cultivars for nodulation and N accumulation were apparent at 25°C, but were abolished by growth at 19°C, indicating that, in spite of differences in growth potential between the cultivars under optimum RZT, both cultivars were equally limited by low RZT. Differences between B. japonicum strains were consistent across temperatures and were largely attributable to higher rates of specific nodule activity recorded for strain 532C, which seemed well adapted to low RZT. These results suggest that the host plant mediates the sensitivity of N2-fixation under low RZT and that inoculation with B. japonicum strains from cold environments is unlikely to enhance soybean N2-fixation under cool soil conditions. 相似文献
33.
Suspension cultures of Sf-9 cells at different stages of growth were infected with a recombinant baculovirus expressing -galactosidase, using a range of multiplicities of infection (MOI) of 0.05 to 50. Following infection, the cells were resuspended either in the medium in which they had been grown or in fresh medium. Specific -galactosidase yields were not markedly affected by either MOI or medium change in cultures infected in early exponential phase (3×106 cells mL–1). In cultures infected at later growth stages, -galactosidase yields could only be maintained by medium replacement. The possibility that this requirement for medium replacement is due either to the accumulation of an inhibitory byproduct or nutrient limitation was examined. Alanine, a major byproduct of cultured insect cell metabolism, did not significantly reduce recombinant protein yield when added to infected cultures in concentrations of up to 40 mM. Following a factorial design, various nutrient concentrates were added alone or in combination to cultures infected in late exponential phase. Additions that included both yeastolate ultrafiltrate and an amino acid mixture restored specific -galactosidase yields to levels observed at earlier growth stages or in late stages with medium replacement; the addition of these concentrates, by permitting production at higher cell density, led to increases in the volumetric yield of recombinant protein. Together or separately, the concentrates when added to uninfected late exponential phase cultures, lead to a doubling of the maximum total cell protein level normally supported by unamended medium. 相似文献
34.
Fluorescence and reflectance characteristics of manganese deficient soybean leaves: Effects of leaf age and choice of leaflet 总被引:5,自引:0,他引:5
Leaf reflectance and fluorescence characteristics of soybean (Glycine max cv Bragg) are influenced strongly by Mn availability. This report evaluates the effects of leaflet choice, leaf age, and leaf nodal position on several spectral characteristics. Leaves were obtained from soybeans grown hydroponically under controlled environmental conditions with wide differences in Mn supply. The ratio of constant yield fluorescence (Fo) to variable yield fluorescence (Fv), the ratios of reflectance at 750 nm to 550 nm and that at 650 nm to 550 nm, the position of the "red edge" near 700 nm, and an index of leaf "yellowness" were measured periodically. Increasing leaf age caused increases in the "red edge" and in both reflectance ratios. Leaf "yellowness" and the fluorescence ratio Fo/Fv decreased with leaf age and increased with leaf nodal position, primarily in Mn deficient leaves. Effects arising from leaf choice were smaller than those caused by Mn deficiency. 相似文献
35.
W. A. Parrott J. N. All M. J. Adang M. A. Bailey H. R. Boerma C. N. Stewart Jr. 《In vitro cellular & developmental biology. Plant》1994,30(3):144-149
Summary Lepidopteran insects are major defoliating pests of soybean in the southeastern United States. Soybean plants transgenic for
a nativecryIA(b) gene fromBacillus thuringiensis var.kurstaki HD-1 were obtained. Embryogenic cultures were induced by plating cotyledons on a Murashige and Skoog-based medium supplemented
with 40 mg/liter of 2,4-dichlorophenoxyacetic acid (2,4-D). The embryogenic cultures were maintained in liquid medium containing
5 mg/liter 2,4-D. These cultures were subjected to microprojectile bombardment, followed by selection on 50 mg/liter hygromycin.
Resistant embryogenic cell lines were transferred to growth regulator-free medium to permit recovery of mature somatic embryos.
After a desiccation period, the somatic embryos were returned to growth regulator-free medium for conversion into plants.
Southern hybridization analysis verified transformation. Feeding assays of T1 plants from one cell line deterred feeding, development, and survival of velvetbean caterpillar at a level comparable to
that of GatIR81-296, a soybean breeding line with a high level of insect resistance. Reduced feeding on T1 plants correlated with the presence of the transgene. 相似文献
36.
Catherine Jumarie Christiane Malo 《In vitro cellular & developmental biology. Animal》1994,30(11):753-760
Summary Caco-2 cell human colon adenocarcinoma cell line was used to study the hormonal regulation of small intestinal epithelial
cell differentiation. We had previously shown that insulin-transferrin-selenium and triiodothyronine (5 × 10−8
M)-supplemented medium can best replace serum after 2 days of culture for both the maintenance and differentiation of Caco-2
cells. The present study demonstrates that precoating petri dishes with complete serum allows the growth and differentiation
of Caco-2 cells seeded directly in serum-free medium. On the other hand, precoating with dialyzed serum inhibits alkaline
phosphatase and dipeptidyl-dipeptidase IV activities by more than 50%. The results obtained with complete serum-precoated
culture plates indicate that there is no synergy between insulin and triiodothyronine because cells maintained in transferrin-selenium
and triiodothyronine-supplemented medium, with or without insulin, express comparable enzyme activities. Moreover, large increases
in alkaline phosphatase and dipeptidyl-dipeptidase IV activities were observed when triiodothyronine was added to the culture
medium by the time confluency was reached. In contrast, γ-glutamyltransferase was lowered to a greater extent when triiodothyronine
was present from the beginning of culture. These findings show that triiodothyronine preferentially stimulates alkaline phosphatase
and dipeptidyl-dipeptidase IV activities during the differentiation period whereas it selectively inhibits γ-glutamyltransferase
during the proliferation phase. Triiodothyronine acts in a dose-dependent manner. 相似文献
37.
M. Kondo W. E. Finkbeiner J. H. Widdicombe 《In vitro cellular & developmental biology. Animal》1993,29(1):19-24
Summary Tracheal epithelial cells were grown on Nuclepore filters coated with human placental collagen. When grown immersed in medium
containing fetal bovine serum, cells displayed an undifferentiated ultrastructure (no cilia and a cell height of ∼ 10 μm).
Short-circuit current (Isc) was approximately 1/10 that of the native epithelium. By contrast, when grown in hormonally defined, serum-free medium with
an air interface, cells showed Isc equal to or greater than the original tissue, possessed cilia, and had a cell height of ∼ 50 μm. Responses in Isc to mediators were similar to those of the original tissue, but differed from those of dog or human tracheal epithelium. Given
the ready availability and low cost of the native tissues, bovine tracheal cultures grown in serum-free medium with an air
interface should prove useful in studies of airway epithelial physiology. 相似文献
38.
Growth of cells in a new defined protein-free medium 总被引:1,自引:0,他引:1
Kjell Bertheussen 《Cytotechnology》1993,11(3):219-231
The development of a new stable synthetic serum replacement (SSR) is described, which allows the cultivation of mammalian cells in a defined, protein-free medium containing only dialyzable components. With a low concentration of insulin (RPMI-SR2 medium), growth rates of the transformed cell lines L929, HELA S3, and the hybridoma 1E6 were comparable to growth rates obtained with a serum-containing medium. The same medium also supported long-term cultivation of non-dividing mouse macrophages. The main principle of SSR is a metal ion buffer containing a balanced mixture of iron and trace metals. Stability against precipitation of important metals is achieved by the combined use of EDTA and citric acid as chelating agents. Efficient iron supply is mediated through the inclusion of the compound Aurintricarboxylic acid as a synthetic replacement for transferrin. SSR also contains a growth-promoting surfactant, Pluronic F68. Thus SSR provides a general foundation for growth and differentiation normally provided by serum.Limitations of other serum-free medium designs are discussed here: 1) the inability of transferrin to chelate all metals in the medium; and 2) the use of inorganic iron salts or iron citrate as an iron supplement leads to rapid precipitation of iron hydroxide in the medium. Both these problems are solved in the design of SSR. 相似文献
39.
James A. Bunce 《Physiologia plantarum》1992,86(1):173-179
The short-term stimulation of the net rate of carbon dioxide exchange of leaves by elevated concentrations of CO2 usually observed in C3 plants sometimes does not persist. Experiments were conducted to test whether the patterns of response to the environment during growth were consistent with the hypotheses that photosynthetic adjustment to elevated CO2 concentration is due to (1) feedback inhibition or (2) nutrient stress. Soybean [Glycine max (L.) Merr. cv. Williams] and sugar beet (Best vulgaris L. cv. Mono Hye-4) were grown from seed at 350 and 700 μl? CO2, at 20 and 25°C, at a photon flux density of 0.5 and 1.0 mmol m?2 S?1 and with three nutrient regimes until the third trifoliolate leaf of soybean or the sixth leaf of sugar beet had finished expanding. Net rates of CO2 exchange of the most recently expanded leaves were then measured at both 350 and 700 μl 1?1 CO2. Plants grown at the elevated CO2 concentration had net rates of leaf CO2 exchange which were reduced by 33% in sugar beet and 23% in soybean when measured at 350 μl 1?1 CO2 and when averaged over all treatments. Negative photosynthetic adjustment to elevated CO2 concentration was not greater at 20 than at 25°C, was not greater at a photon flux density of 1.0 than at 0.5 mmol m?2 S?1 and was not greater with limiting nutrients. Furthermore, in soybean, negative photosynthetic adjustment could be induced by a single night at elevated CO2 concentration, with net rates of CO2 exchange the next day equal to those of leaves of plants grown from seed at the elevated concentration of CO2. These patterns do not support either the feedback-inhibition or the nutrient-stress hypothesis of photosynthetic adjustment to elevated concentrations of CO2. 相似文献
40.
Adle Martial Isabelle Gaillard Jean-Marc Engasser Annie Marc 《Enzyme and microbial technology》1995,17(12):1062-1066
A homemade serum-free medium containing a low protein level under 0.1 g l−1 has been proved to support long-term cultures of VO 208 hybridoma cells successfully up to 50 days. The low protein level was achieved by supplying the lipids through liposomes containing cholesterol, oleic acid,
- dipalmitoyl phosphatidylcholine, and bovine serum albumin. The influence of the liposome content in the feeding medium was studied in a continuous culture performed with step variations of the liposomes level, from 7.5 to 30 ml l−1. The cell density decreased at the highest liposomes content while it became higher with 7.5 or 12 ml l−1 of liposomes. For each step variation appeared a transitory activation of the specific rates of nutrient consumption, metabolite production and antibody secretion, as well as a transitory decrease of the specific cell growth rate. The overall structure of the antibodies was not affected during the culture. 相似文献