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1.
Fabienne Kinard Lieve de Clercq Brigitte Billen Brigitte Amory Jules-Joseph Hoet Claude Remacle 《In vitro cellular & developmental biology. Plant》1990,26(10):1004-1010
Summary Cell suspensions prepared by collagenase digestion of pancreata obtained from 21.5-d-old rat fetuses were preincubated in
RPMI medium containing 10% fetal bovine serum (FBS), to ensure cell adhesion. Twenty hours later, this medium was replaced
by a chemically defined medium. Dulbecco's modified Eagle's (DME)-F12 was used alone or supplemented with various combinations
of transferrin, sodium selenite, or Ultroser G. The evolution of the culture and the islet ultrastructure were similar in
defined and serum-containing media. However, in the defined medium, the neoformed islets seemed less numerous, and the fibroblast
layer less dense, when compared to the RPMI+10% FBS control medium. At Day 7, in defined media, the total insulin content
per dish was half that of control cultures. None of the tested additives improved the yield of the cultures. The fractional
insulin release per day was elevated in defined media. In subsequent incubations, glucose and leucine stimulated insulin release
in a way characteristic of these cells of fetal origin. The labeling index of islet cells cultured in DME-F12 reached 10.7%,
which is not far from that observed in RPMI+10% FBS. Such a defined medium is useful to study B cell physiology, avoiding
the possible interaction of serum components with substances to be tested.
The support of the Fonds National de la Recherche Scientifique and the ‘Loterie Nationale’ of Belgium is also acknowledged. 相似文献
2.
Marc E. Bracke Marc De Mets Rita M. L. Van Cauwenberge Luc Vakaet Jr. Georges K. De Bruyne Marc M. Mareel 《In vitro cellular & developmental biology. Plant》1986,22(9):508-514
Summary Confronting cultures of precultured embryonic chick heart fragments (PHF) with aggregates of malignant cells in vitro have
been shown to be relevant for a number of aspects of tumor invasion in vivo. Preculture of the heart fragments, formation
of cell aggregates and subsequent culture of confronting pairs have so far been done only in serum-containing culture media.
We describe here confronting cultures of PHF with invasive MO4 mouse cell aggregates or noninvasive MDCK dog kidney cell aggregates in serum-free media. Heart fragments precultured in
the absence of serum seemed to be necrotic after confronting culture in serum-free media. However, preculturing in media supplemented
with 10% fetal bovine serum allowed us to do subsequent confronting cultures in absence of serum. Cell aggregates were also
prepared in serum-containing medium. MO4 cells occupied and replaced the heart tissue within 4 d, whereas MDCK cells remained at the periphery, of the PHF. This indicates
that serum-free confronting cultures can discriminate between invasive and noninvasive cells. The viability of individual
PHF and cell aggregates cultured in the same way as in confrontations was ascertained by histology and by explantation and
postculturing on a solid tissue culture substrate. Growth of the cultures was smaller in serum-free media than in media supplemented
with 10% fetal bovine serum. The main advantage of serum-free culture conditions in vitro is the elimination of the influence
of serum components on invasion, and the ability to examine the effect on invasion of drugs that are, susceptible to inactivation
by serum.
This work was supported by the Fonds van de Sport Vereniging tegen de Kanker, Brussels, Belgium, and the Fonds voor Geneeskundig
Wetenschappelijk Onderzoek Brussels, Belgium 相似文献
3.
R. Loppes 《Molecular & general genetics : MGG》1969,104(2):172-177
Summary Forward mutations to auxotrophy have been induced in Chlamydomonas reinhardi after treatment of wild-type cells with ethyl methanesulfonate (EMS) and plating on a selective medium supplemented with yeast extract but deprived of ammonium chloride.Among the 41 stable mutants isolated, 6 were arginine-requiring. Reconstruction experiments show that the growth of all these arginine auxotrophs is inhibited on media containing high concentrations of NH
4
+
ions. It is concluded that the isolation of this new class of mutants was made possible because of the low ammonium concentration of the plating medium.It appears therefore that in Chlamydomonas, as in other microorganisms, the specificity of mutations may be imposed by the selective medium.Chargé de Recherches du Fonds National de la Recherche Scientifique. 相似文献
4.
Marie-France Grasset Jean Paul Blanchet 《In vitro cellular & developmental biology. Plant》1984,20(4):302-304
Summary The growth of late erythroid precursors (CFU-Es) from adult bone marrow is inhibited when Iscove's modified Dulbecco's medium
supplied in liquid form is used. Catalase and other H2O2 destroying compounds restore the capacity of culture medium to support colony development. However early precursors from
adult bone marrow and fetal liver CFU-Es were resistant to H2O2.
This work was supported by grants from the Centre National de la Recherche Scientifique, the Institut National de la Santé
et de la Recherche Médicale and the Fédération Nationale des Centres de Lutte contre le Cancer. 相似文献
5.
Christine Halleux Yves-Jacques Schneider 《In vitro cellular & developmental biology. Animal》1991,27(4):293-302
Summary Iron absorption by intestinal epithelial cells, passage onto plasmatic apotransferrin, and regulation of the process remain
largely misunderstood. To investigate this problem, we have set up an in vitro model, consisting in CaCo2 cells (a human colon
adenocarcinoma line, which upon cultivation displays numerous differentiation criteria of small intestine epithelial cells).
Cells are cultivated in a serum-free medium, containing 1μg/ml insulin, 1 ng/ml epidermal growth factor, 10μg/ml albumin-linoleic acid, 100 nM hydrocortisone, and 2 nM T3 on new, transparent, Cyclopore polyethyleneterephthalate microporous membranes coated with type I collagen. Cells rapidly
adhere, grow, and form confluent monolayers; after 15 days, scanning electron microscopy reveals numerous uniform microvilli.
Domes, which develop on nonporous substrata, are absent on high porosity membranes. Culture medium from upper and lower compartments
of microplate inserts and cell lysates were immunoprecipitated after labeling with [3H]glucosamine and leucine; analysis was done by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed
by autoradiography. [3H]transferrin is found mainly in the lower compartment and in cells; [3H]apolipoprotein B is released in both compartments, and fibronectin almost entirely recovered in the lower compartment; [3H]transferrin receptors and ferritin are only present in cell lysates. Binding experiments also show that transferrin receptors
are accessible from the lower compartment. These results suggest that CaCo2 cells, cultivated in synthetic medium on membranes
of appropriate porosity, could provide an in vitro model of the intestinal barrier, with the upper compartment of the culture
insert corresponding to the apical pole facing the intestinal lumen and the lower one to the basal pole in contact with blood.
This work was supported by the Belgian Fonds de la Recherche Scientifique Médicale (grant 3.4551.88) and the Walloon Region. 相似文献
6.
R. D. Ellender J. H. Wharton B. L. Middlebrooks 《In vitro cellular & developmental biology. Plant》1979,15(2):112-113
Summary A cell line, SP-2, was established from spleen tissue ofBairdiella chrysura (the silver perch). The line is susceptible to lymphocystis virus and the amphibian LT-1 virus but refractory to six additional
viruses. The modal chromosome number of primary silver perch cells is 48, but SP-2 cells are heteroploid. For growth, Leibovitz
L-15 medium supplemented with fetal bovine serum and sodium chloride (to 0.150m) was employed. Cells replicated best at 25° to 28° C.
Funds for this investigation were supplied by the National Oceanographic and Atmospheric Administration, Office of Sea Grant
No. 04-3-158-58. 相似文献
7.
Roeland van Wijk Lydie Tichonicky Jaques Kruh 《In vitro cellular & developmental biology. Plant》1981,17(10):859-862
Summary Exposure of HTC cells to sodium butyrate caused inhibition of growth. The site of growth inhibition was studied by time-lapse
cinematography and [3H]thymidine incorporation studies. Evidence is presented that sodiunm butyrate affected the cell cycle at a specific point
immediately after mitosis. Inasmuch as it does not modify the interphase duration after its removal, butyrate may be used
for HTC synchronization.
This work was supported by l'Institut Nationale de la Santé et de la Recherche Médicale and la Centre Nationale de la Recherche
Scientifique (L. T. and J. K.). 相似文献
8.
A myogenic cell line with altered serum requirements for differentiation 总被引:19,自引:0,他引:19
Dfferentiation properties of a cell line, L84, which originated from a non-fusing clone isolated from the myogenic line L8, are described. In nutritional medium supplemented with 10% serum used routinely with L8 cells, L84 cells continue to proliferate to very high densities and fail to form multinucleated fibres. When grown in medium supplemented with 2% horse serum of 2% horse serum plus 0.1% microng/ml insulin, L84 cells behave very similarly to L8 cells grown in medium supplemented with 10% horse serum: when the cultures reach confluency, proliferation decreases and cells start to fuse and form a dense network of fibres. Large increases in creatine kinase activity and synthesis of myosin are associated with cell fusion. Under conditions in which L84 cells do not fuse the increase in these synthetic activities is not observed, even after extremely high cell densities are reached. The data show that L84 cells retain the programme for their differentiation into muscle fibres. The difference between L84 and its progenitor line L8 lies in the sensitivity to the environmental conditions which trigger the expression of this programme. 相似文献
9.
Summary. The nature of pectins (acidic, methyl-, or acetyl-esterified) in the shoot meristem of Sinapis alba was assessed by immunocytochemistry with the 2F4 monoclonal antibody in light and electron microscopy. This antibody is specific
for “egg-boxes” – the polygalacturonic acid conformation induced by calcium as described in Liners et al. (Plant Physiol.
99: 1099–1104, 1992). Hardly any acidic pectin was detected in meristem walls; the pectins were largely methyl-esterified
and esterified by acetyl groups and/or other esters. After in situ chemical or enzymatic de-esterification, labeling was distributed
over the primary wall and the middle lamella of meristematic cells. Acidic pectin and Ca2+-cross-linked homogalacturonans were absent from the pit fields, where plasmodesmata traverse the middle lamella. The type
and distribution of pectins are discussed in relation to cellular adhesion between active meristem cells.
Correspondence and reprints: Unité de Recherches en Biologie Cellulaire Végétale, Département de Biologie, Facultés Universitaires
Notre-Dame de la Paix, rue de Bruxelles 61, 5000 Namur, Belgium. 相似文献
10.
We report here that it is possible to induce differentiation in a subline of L5 myoblast line (L5/A10) by manipulating the culture media. When L5/A10 myoblast are cultured in F14 supplemented with 10% fetal calf serum the cells grow with a division time of 12 h and reach confluency at a cell density of approximately 2.4 X 10(5) cells per cm2, without undergoing differentiation, characterized, morphologically, by formation of multinucleated fibers, and biochemically, by the synthesis of muscle specific proteins such as creatine phosphokinase or myokinase. However, cells, grown in F14 + 10% fetal calf serum, will undergo regular differentiation after a limited number of division when transferred to F14 medium supplemented with limiting concentrations (1-2%) of fetal calf serum. Investigations of the biochemistry of myoblast differentiation in cell culture will be facilitated by the availability of a cell line that can undergo differentiation under controlled conditions. 相似文献
11.
The glutamine requirement for thein vitro proliferation of fish cells was investigated with cell lines from four different species and three tissues: goldfish skin (GFSk-S1), Chinook salmon embryo (CHSE-214), and raibow trout liver (RTL-W1) and spleen (RTSp-W1). With a supplement of fetal bovine serum, the basal medium, Leibovitz's L-15, without glutamine supported the proliferation of all four cell lines as well, or nearly as well, as L-15 with 2 mM glutamine. This was true over short term assays of two to four weeks and for continuous propagation. CHSE-214 also grew as well with or without 2 mM glutamine in Minimum Essential Medium with fetal bovine serum. However, when the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in L-15 without glutamine. Therefore, glutamine was not required for growth in L-15, and in fact, was inhibitory in the absence of the dialyzable fraction of serum. By contrast, glutamine appeared to be important for growth in Minimum Essential Medium. When the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in Minimum Essential Medium with 2 mM glutamine. These results suggest that the glutamine requirement for thein vitro proliferation of fish cells is conditional and depends on the basal medium and serum supplement.Abbreviations BSA
bovine serum albumin
- CHSE-214
Chinook samon embryo cell line
- dFBS
dialyzed fetal bovine serum
- FBS
fetal bovine serum
- GFSk-S1
goldfish skin cell line
- GS
glutamine synthetase
- L-15
Leibovitz's L-15 media
- L929
mouse fibroblast cell line
- MEM
minimum essential medium Eagle
- PBS
phosphate buffered saline
- RTL-W1
rainbow trout liver cell line
- RTSp-W1
rainbow trout spleen cell line 相似文献
12.
A marine fish cell line from the snout of red spotted grouper Epinephelus akaara, a protogynous hermaphrodite, was established, characterized, and subcultured with more than 60 passages. The grouper snout
cell line (GSC) cells multiplied well in Dulbecco’s modified Eagle’s medium (DMEM) medium supplemented with 10% fetal bovine
serum. The optimal growth temperature was 25°C, and morphologically the cells were fibroblastic. Chromosome analysis revealed
that the GSC cell line has a normal diploid karyotype with
. A virus titration study indicated that the cells were susceptible to turbot Scophthalmus Maximus rhabdovirus (SMRV) (108.5 TCID50 ml−1), while the viral titer of frog Rana grylio virus 9807 (RGV9807) reached 103.5 TCID50 ml−1. The infection was confirmed by cytopathic effect (CPE), immunofluorescence, and electron microscopy experiments, which detected
the viral particles in the cytoplasm of virus-infected cells, respectively. Further, significant fluorescent signals were
observed when the GSC cells were transfected with pEGFP vector DNA, indicating their potential utility for transgenic and
genetic manipulation studies. 相似文献
13.
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
14.
Hemolymph analysis and evaluation of newly formulated media for culture of shrimp cells (Penaeus stylirostris) 总被引:1,自引:0,他引:1
Shimizu C Shike H Klimpel KR Burns JC 《In vitro cellular & developmental biology. Animal》2001,37(6):322-329
Summary Creation of a shirmp cell line has been an elusive goal. This failure may be due to the composition of the cell culture medium,
which may be inadequate to support primary cultured cells. Shrimp hemolymph should contain the nutritional components needed
to support cell growth and division. We report here the comprehensive biochemical analysis of hemolymph from the blue shrimp,Penaeus stylirostris (Litopenaeus stylirostris) (see Holthuis, L. B. Shrimps, and prawns of the world, in: FAO species catalog. Vol. 1. Rome: Food and Agriculture Organization
of the United Nations; 1980), for free amino acids (FAAs), carbohydrates, electrolytes, metals, pH, and osmolality. Levels
of hemolymph components were compared to 2×L-15 with 20% fetal bovine serum, a commonly used culture medium for crustacean
cells. The FAAs, taurine and proline, and the metals, strontium and zinc, were significantly higher in hemolymph than in the
2×L-15 medium. In contrast, other FAAs were up to 50 times higher in the 2×L-15 medium than in the hemolymph. To mimic more
closely the hemolymph composition, we created two new media based on either the 0.2×L-15 or the M199 medium. We compared the
microscopic appearance of cells cultured in these media and evaluated deoxyribonucleic acid (DNA) and protein synthesis by3H-thymidine uptake and35S-methionine uptake assays. The ovary cells ofP. stylirostris cultured in either of the new media formed monolayers, while the cells cultured in 2×L-15 medium did not. Despite these differences,
there was no evidence of sustained DNA or protein synthesis with any of the media. Future studies to establish a shrimp cell
line should focus on analysis of the cell cycle and on overcoming the molecular blocks to cell division. 相似文献
15.
Ferrous ion (Fe2+) has been considered to be a cause of neuronal oxidative injury. Since body fluids contain protein and serum is an essential component of tissue culture medium, we have examined the role of serum protein on Fe2+-mediated oxidative stress using PC12 cells and rat cerebral cortices. Fe2+ or the combination of ascorbate and Fe2+ increased concentrations of thiobarbituric acid reactive substances (TBARS) in PC12 cells and cerebrocortical homogenates in medium (RPMI 1640), but did not increase TBARS when the medium was supplemented with 10% fetal bovine serum. Treatment with ascorbate/Fe2+ in serum-free medium reduced endogenous glutathione (GSH) concentration in PC12 cells. However, the medium supplemented with serum did not reduce GSH concentrations. PC12 cell death induced by ascorbate/Fe2+ was alleviated by increasing serum or bovine albumin concentrations in the medium. These observations indicated that oxidative injury caused by the transition metal ion could be lessened by adding fetal bovine serum to culture medium. 相似文献
16.
Summary We describe the in vitro influence of 3,5,3′-triiodo-l-thyronine (T3),l-thyroxine (T4), a thyroid-stimulating hormone (TSH), and/or estradiol (E2: chosen as the control of the methodology) on the cell kinetics (cell distribution in the S+G2+M phases) of mouse MXT and human MCF-7 mammary cancer cells. Experiments were performed by means of a cell image processor,
analyzing MCF-7 or MXT cells that had been grown on glass cover slips and whose nuclei had been stained by the Feulgen reaction,
which is selective and quantitative (stoichiometric) with respect to DNA. We show that T3, T4, and TSH at 0.01 μM dramatically stimulate the cell kinetics of the MXT mouse and the MCF-7 human mammary cancer cell lines. Indeed, the three
hormones bring about a significant transient increase in the S+G2+M fraction as does E2. Furthermore, our data indicate that E2 and TSH are antagonistic with regards to MXT or MCF-7 cell kinetics.
This work is supported by grants awarded by the IRSIA and the Fonds de la Recherche Scientifique Médicale (FRSM, Belgium). 相似文献
17.
P. M. Simon M. Kedinger F. Raul J. F. Grenier K. Haffen 《In vitro cellular & developmental biology. Plant》1982,18(4):339-346
Summary Jejunal mucosa of 6 d-old rats were cultured for 24 and 48 h in the presence of thyroxine, insulin, pentagastrin, glucagon,
epidermal growth factor (EGF) or dibutyryl-A-3:5-MP cyclic with or without dexamethasone (DX). The enzymes were assayed on
the purified brush borders. The various agents added alone to the basic culture medium had no effect with the exception of
DX on the levels of enzyme activities. Dexamethasone alone induced sucrase, stimulated maltase, and protected other brush
border enzyme activities (aminopeptidase, lactase, and alkaline phosphatase). When added to DX-supplemented medium, only the
following factors modified the levels of enzymatic activities observed with DX alone. Insulin (10−6
M) increased maltase, alkaline phosphatase, and lactase activity to a greater extent than DX at 24 h culture, the effect being
maintained at 48 h on alkaline phosphatase only. At 48 h culture, both EGF (10−8
M) and dbcAMP (10−3
M) decreased DX-induced sucrase activity. The latter agent also depressed DX-stimulated aminopeptidase activity.
This work was supported by the Institut National de la Santé et de la Recherche Médicale, Centre National de la Recherche
Scientifique, and a grant 79.7.1243 from the Délégation Générale a la Recherche Scientifique et Technique.
P. M. S. is a recipient of a grant from Fondation de la Recherche Médicale (France). 相似文献
18.
Dewitte-Orr SJ Lepic K Bryson SP Walsh SK Lee LE Bols NC 《In vitro cellular & developmental biology. Animal》2006,42(8-9):263-272
Summary A continuous cell line, PBLE, was developed from the adherent cells in a culture of peripheral blood leukocytes from the American
eel, Anguilla rostrata. The cells were grown in Leibovitz's L-15 basal medium supplemented with 20% fetal bovine serum (FBS). Under normal culture
conditions at 18° C, the morphology of PBLE was fibroblast-like. The cultures have been subcultured over 80 times and have
been cryopreserved successfully. These cells have a diploid karyotype of 38 chromosomes, survived temperatures from 5 to 36°
C, and proliferated at temperatures from 5° C to at least 30° C. PBLE underwent apoptosis in response to gliotoxin, but did
not show a respiratory burst. Results suggest that PBLE may have arisen from a circulating mesenchymal stem cell. PBLE was
susceptible to Chum salmon reovirus (CSV) and supported CSV replication. Therefore this cell line should be useful in studying
eel specific virus-host interactions. 相似文献
19.
Kamita SG Do ZN Samra AI Hagler JR Hammock BD 《In vitro cellular & developmental biology. Animal》2005,41(5-6):149-153
Summary Four continuous cell lines were established from the embryos of the glassy-winged sharpshooter, Homalodisca coagulata (Say), an economically important insect vector of bacterial pathogens of grape, almond citrus, oleander, and other agricultural
and ornamental plantings. The cell lines were designated GWSS-Z10, GWSS-Z15, GWSS-G3, and GWSS-LH. The GWSS-Z10, GWSS-Z15,
and GWSS-G3 lines were cultured in Ex-Cell 401 medium supplemented with 10% fetal bovine serum (FBS), whereas the GWSS-LH
line was cultured in LH medium supplemented with 20% FBS. The cell lines were characterized in terms of their morphology,
growth, protein composition, and polymerase chain reactionamplification patterns of their chromosomal deoxyribonucleic acid.
The population doubling times of GWSS-Z10, GWSS-Z15, GWSS-G3, and GWSS-LH were 46.2, 90.9, 100.3 and 60.2 h, respectively.
These lines should be useful for the study of insect-pathogenic viruses of leafhoppers, aphids, treehoppers, and other related
insects as well as plant-pathogenic viruses that are transmitted by these insects. 相似文献
20.
W. -H. Tsai G. L. Holbrook C. Schal A. -S. Chiang 《In vitro cellular & developmental biology. Animal》1995,31(7):542-546
Summary An in vitro organ culture system was established to support growth of corpora allata from the cockroachDiploptera punctata. During a 1-wk incubation in L-15B medium supplemented with 10% fetal bovine serum (FBS) and 10% cockroach hemolymph, adult
male corpora allata exhibited a cycle of de novo DNA synthesis followed by cell division. The number of S-phase cells and
metaphase cells per corpus allatum were counted from whole-mount monolayers after labeling in vitro with 5′-bromo-2′-deoxyuridine
and exposure to colchicine, respectively. While both FBS and cockroach hemolymph were essential for proliferation of allatal
cells, the growth-promoting effect of insect hemolymph was not species-specific and adult female hemolymph was more potent
than hemolymph from adult males. Furthermore, DNA synthesis of corpus allatum cells was stimulated in vitro by 20-hydroxyecdysone.
This sensitive assay system will be of immense utility in the search for allatal growth factors. 相似文献