Encephalomyelitis vaccines: a Vero-derived cell culture alternative to primary duck embryonic cell cultures

Vaccine ◽  
1988 ◽  
Vol 6 (1) ◽  
pp. 6
Author(s):  
J.B. Katz ◽  
S.K. Hanson
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Embryonic Cell

Obtaining and Characterization of Alhagi persarum Boiss. et Buhse Callus Cell Cultures, Isoflavonoid Producers

2020 ◽  
Vol 36 (6) ◽  
pp. 35-48
Author(s):  
D.V. Коchkin ◽  
G.I. Sobolkovа ◽  
А.А. Fоmеnkov ◽  
R.А. Sidorov ◽  
А.М. Nоsоv
Keyword(s):  
Fatty Acids ◽  
Cell Culture ◽  
Liquid Chromatography ◽  
Secondary Metabolites ◽  
Cell Cultures ◽  
Mass Spectrometric ◽  
Mass Spectrometric Detection ◽  
Gas Liquid Chromatography ◽  
Callus Cell

The physiological characteristics of the callus cell cultures of Alhagi persarum Boiss et Buhse, a member of the legume family, widely used in folk medicine, have been studied. It was shown that the source of the explant was an important factor in the initiation of callusogenesis: more intense callusogenesis (almost 100%) was observed for explants from various organs of sterile seedlings, rather than intact plants (less than 30%). As a result, more than 20 lines of morphologically different callus cell cultures were obtained, and the growth parameters for the 5 most intensively growing lines were determined. The composition of fatty acids (FA) of total lipids and secondary metabolites in the most physiologically stable callus line Aр-207 was analyzed. Using capillary gas-liquid chromatography with mass spectrometric detection (GLC-MS), 19 individual C12--C24 FAs were identified, the main fraction of which were palmitic (~ 23%), stearic (~ 22%), linoleic (~ 14%) and α-linolenic (~ 33%) acids. The established atypical ratio of FAs (a simultaneous high content of both saturated FAs and polyunsaturated α-linolenic acid) is possibly due to the adaptation of cells to in vitro growth conditions. Phytochemical analysis of the secondary metabolites was carried out using ultra-performance liquid chromatography with electrospray ionization mass spectrometric detection (UPLC MS). Compounds belonging to different structural groups of isoflavones were found. Aglycones (calycosin, formononetin and afrormosin isomer), glucosides (formononetin glucoside), as well as esters of glucosides (malonylglycosides of calicosin, formononetin, afrormosin isomers, glycitein and genistein) were detected. These secondary metabolites are widespread in plants of the Fabaceae family; however, isoflavones are rare in representatives of the Alhagi genus. The presence of malonylated isoflavone glycosides in Alhagi spp. was shown for the first time. endemic plant species, Alhagi, in vitro cell culture, callus cell culture, isoflavones, fatty acids All studies were carried out using the equipment of the "Experimental Biotechnological Facility" and the "All-Russian Collection of Cell Cultures of Higher Plants" of IРР RAS. This work was supported by the Russian Foundation for Basic Research (RFBR), contract no.18-54-06021 (Az_a), and the Government of the Russian Federation, Megagrant Project no. 075-15-2019-1882.

scholarly journals Isolation of Porcine Circovirus-like Viruses from Pigs with a Wasting Disease in the USA and Europe

1998 ◽  
Vol 10 (1) ◽  
pp. 3-10 ◽  
Author(s):  
G. M. Allan ◽  
F. McNeilly ◽  
S. Kennedy ◽  
B. Daft ◽  
J. A. Ellis ◽  
...  
Keyword(s):  
Cell Culture ◽  
Lymph Node ◽  
Nucleic Acid ◽  
Cell Cultures ◽  
Virus Isolation ◽  
Chicken Anemia Virus ◽  
Porcine Circovirus ◽  
Tissue Homogenate ◽  
Mesenteric Lymph ◽  
Viral Particles

Samples of lung, liver, kidney, pancreas, spleen, and lymph node from pigs with postweaning multisystemic wasting syndrome from California (USA) and samples of mesenteric lymph nodes from similarly diseased pigs from Brittany (France) were examined by light microscopy, in situ hybridization (ISH), and/or virus isolation. Whole genomic probes for porcine circovirus (PCV) and chicken anemia virus (CAV) were used for ISH. Tissue homogenate supernatants were inoculated onto PK/15 cells for virus isolation, and the presence of viral antigen and viral particles was verified by indirect immunofluorescence, ISH, and electron microscopy. Histologic examination of lung from pigs from California revealed interstitial pneumonia, alveolar epithelial hyperplasia, and basophilic nuclear and cytoplasmic inclusions in mononuclear cell infiltrates and various pulmonary epithelial cells. Granulomatous lymphadenitis with syncytial cells typified the lesions seen in the pigs from France. PCV-like nucleic acid was detected by ISH in lung, pancreas, lymph node, kidney, and liver in pigs from California. Positive signal was also obtained in lymph node sections from pigs from France. Probes for CAV were consistently negative. PK/15 cell cultures inoculated with lung preparations from diseased California pigs and mesenteric lymph node preparations from pigs from France had positive fluorescence by indirect staining for PCV using pooled polyclonal pig sera and hyperimmune rabbit serum and had variable staining with a panel of 7 monoclonal antibodies specific for cell culture contaminant PCV. PCV-like nucleic acid was also detected by ISH in cell cultures. Cytopathic effect was not observed Electron microscopic examination of inoculated cell cultures revealed 17-nm viral particles morphologically consistent with PCV No other virus particles were observed. Although genomic analysis for the definitive identification of these viral isolates remains to be done, the evidence provided strongly suggests that these tissue isolates are closely related to, although antigenically distinct from, the original PCV cell culture contaminant.

scholarly journals Binary Biocompatible CNC–Gelatine Hydrogel as 3D Scaffolds Suitable for Cell Culture Adhesion and Growth

Applied Nano ◽  
2021 ◽  
Vol 2 (2) ◽  
pp. 118-127
Author(s):  
Luca Zoia ◽  
Anna Binda ◽  
Laura Cipolla ◽  
Ilaria Rivolta ◽  
Barbara La Ferla
Keyword(s):  
Tissue Engineering ◽  
Cell Culture ◽  
Cell Cultures ◽  
Cellulose Nanocrystals ◽  
Potential Application ◽  
Chemical Properties ◽  
Cellular Adhesion ◽  
Chemical Crosslinking ◽  
3D Scaffolds ◽  
Physical Chemical

Binary nano-biocomposite 3D scaffolds of cellulose nanocrystals (CNCs)—gelatine were fabricated without using chemical crosslinking additives. Controlled oxidative treatment allowed introducing carboxyl or carbonyl functionalities on the surface of CNCs responsible for the crosslinking of gelatine polymers. The obtained composites were characterized for their physical-chemical properties. Their biocompatibility towards different cell cultures was evaluated through MTT and LDH assays, cellular adhesion and proliferation experiments. Gelatine composites reinforced with carbonyl-modified CNCs showed the most performing swelling/degradation profile and the most promising adhesion and proliferation properties towards cell lines, suggesting their potential application in the field of tissue engineering.

Antimigratory effect of berberine in T98G, U87MG and primary glioma cell culture.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e15045-e15045
Author(s):  
Irina V. Mezhevova ◽  
Svetlana Yu. Filippova ◽  
Sofia V. Timofeeva ◽  
Anastasia O. Sitkovskaya ◽  
Tatiana V. Shamova ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Lines ◽  
Cell Cultures ◽  
Glioma Cell ◽  
Glioma Cells ◽  
Surgical Removal ◽  
Experimental Sample ◽  
Wound Area ◽  
Migratory Activity ◽  
Primary Glioma

e15045 Background: Berberine is an alkaloid compound with a structure that is highly similar to that of intercalating agents. It affects numerous cell signaling pathways and is widely studied as potential anticancer drug. It is known that berberine affects cancer cells migration through metalloproteinase-2 inhibition, but this effect was never studied on glioma cells. Anti-migratory drugs are of special interest in brain cancer therapy since glioma's highly invasive nature makes total surgical removal of tumor practically impossible. The aim of the study was to evaluate berberine anti-migratory activity on glioma cells. Methods: Cell migration capacity of T98G and U87MG cell lines, as well as primary glioma cell culture established in our laboratory, was assessed via standard wound healing assay with automated image acquisition and analysis on Lionheart FX (BioTek) cell imager. Prior to assay setting up cell cultures were maintained in DMEM medium with L-glutamine (1 μM) (Gibco) and 10% FBS (Gibco) at 37C0 and 5.0% CO2. Cells were seeded at 250 000 cells per well on 24-well plates and incubated overnight in order to attach to plate bottom. After that a vertical wound was made manually in each well, and berberine was added to experimental wells to final concentration 50 mg/L. Plates with cells were continuously incubated and photographed in cell imager at 37C0 and 5.0% CO2. The extent of cells migration was measured as the percent of wound area decrease after 24 hours of incubation in relation to starting time point. Data are given as: Mean ± 95% confidence interval. Results: In our study we berberine exhibited anti-migratory activity in all cell cultures under study. In rather fast growing primary cell culture wound area decrease was 99.23%±0.62% in control sample and 91.75%±0.28% in experimental sample. The difference was small but significant at p < 0.001 level (df = 30). Popular permanent glioma cell lines T98G and U87MG showed more prominent decrease in studied parameter with higher degree of variance at the same time. In T98G wound area decrease was 71.6%±12.3% in control and 48.8%± 7.6% in experimental samples after 24 hours of cultivation in presence of 50 mg/L berberine. While U87MG demonstrated 60.28%±5.13% and 37.5%± 8.34% wound area decrease accordingly. The obtained difference between control and experimental groups in permanent cell cultures was statistically significant at the 0.05 level (df = 30). Conclusions: Our preliminary research proved berberine to be potent anti-migratory agent in glioma treatment. Further investigations are needed to evaluate its ability to inhibit glioma cell expansion in vivo.

Herbicidal Concentrations of Picloram in Cell Culture and Leaf Buds

Weed Science ◽  
1972 ◽  
Vol 20 (2) ◽  
pp. 185-188 ◽  
Author(s):  
F. S. Davis ◽  
A. Villarreal ◽  
J. R. Baur ◽  
I. S. Goldstein
Keyword(s):  
Cell Culture ◽  
Glycine Max ◽  
Gossypium Hirsutum ◽  
Cell Cultures ◽  
Tolerance Limit ◽  
Primary Leaf ◽  
Fresh Weight ◽  
Internal Concentration ◽  
Order Of Magnitude

Cell cultures of soybean(Glycine max(L.) Merrill ‘Acme’) were exposed to media containing 4-amino-3,5,6-trichloropicolinic acid (picloram) for 15 days. Picloram also was supplied once in droplets (water) to cotyledons of 10 to 13-day-old seedlings of cotton(Gossypium hirsutumL. ‘Champion’). The amounts of picloram necessary to reach and exceed the 50% tolerance limit (TL50) of the cell cultures (inhibition) and of the primary leaf buds (toxicity) were established, and internal picloram concentrations then were determined. Internal concentrations at the TL50were 0.17 nM/g fresh weight and 14.7 nM/g fresh weight for cell cultures and leaf buds, respectively. These values are approximately 10−7and 10−5molar. In leaf buds, concentrations increased rapidly for 36 hr after treatment and declined slowly thereafter. Primary leaf buds accumulated up to several times the lethal internal concentration of picloram when the dosage to the cotyledons was increased by one order of magnitude.

scholarly journals Chitosan Elicitation for Enhancing of Vincristine and Vinblastine Accumulation in Cell Culture of Catharanthus roseus (L.) G. Don

2018 ◽  
Vol 10 (12) ◽  
pp. 287
Author(s):  
Pawnpirun Pliankong ◽  
Padungsak Suksa-Ard ◽  
Surawit Wannakrairoj
Keyword(s):  
Molecular Weight ◽  
Cell Culture ◽  
Stationary Phase ◽  
Liquid Medium ◽  
Cell Cultures ◽  
Catharanthus Roseus ◽  
Dry Weight ◽  
Shrimp Shell ◽  
Anti Cancer ◽  
Herbal Plant

Catharanthus roseus (L.) G. Don is an important herbal plant. There are two important alkaloids, vinblastine and vincristine, use in anti-cancer drugs. In this study production of the two alkaloids was enhanced in C. roseus cell cultures, in a Murashige and Skoog (MS) liquid medium supplemented with 1.5 mg/L 2,4-D, 0.5 mg/L kinetin and 30 g/L sucrose, by adding 0, 50, 100, 250 or 500 mg/L medium molecular weight chitosan or chitosan derived from shrimp shell. After 14 days of culture, the cell suspension at stationary phase in the 100 mg/L medium molecular weight chitosan could produce the highest amounts of vinblastine and vincristine at 4.15 and 5.48 &micro;g/mg cell dry weight, respectively. At the same time, the controls (0 mg/L chitosan) produced the two alkaloids at only 2.43 and 2.49 &micro;g/mg cell dry weight, respectively. For chitosan from shrimp shell, it was found that 100 mg/L chitosan could lead to the highest quantity of 4.09 &micro;g vinblastine/mg cell dry weight. The highest amount of 5.47 &micro;g vincristine/mg cell dry weight was obtained when 250 mg/L chitosan was added.

scholarly journals Analysis on the prokaryotic microbiome in females and embryonic cell cultures of Rhipicephalus sanguineus tropical and temperate lineages from two specific localities in Brazil

2021 ◽  
Vol 30 (3) ◽  
Author(s):  
Mayara de Cassia Luzzi ◽  
Lucas Amoroso Lopes de Carvalho ◽  
Daniel Guariz Pinheiro ◽  
Leidiane Lima-Duarte ◽  
Jaqueline Valéria Camargo ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Rhipicephalus Sanguineus ◽  
Distribution Patterns ◽  
Embryonic Cell ◽  
Tick Cell ◽  
Illumina Platform ◽  
Region Gene ◽  
Host Environment ◽  
Canine Monocytic Ehrlichiosis ◽  
Vertebrate Hosts

Abstract Two lineages of Rhipicephalus sanguineus are known in Brazil: the temperate or southern and the tropical or northern populations. The distribution patterns of both lineages of R. sanguineus have epidemiological implications that can affect vectorial competence concerning Ehrlichia canis, the agent of canine monocytic ehrlichiosis. Intending to identify the microbiomes of both lineages and compare microorganisms in R. sanguineus, we used the 16S rRNA (V4-V5 region) gene-based metataxonomic approach, through NGS sequencing on the MiSeq Illumina platform. We selected specimens of females from the environment and samples of primary embryonic cell cultures, from both lineages, and this was the first study to investigate the prokaryotic microbiome in tick cell cultures. The results showed that many bacterial taxa detected in the samples were typical members of the host environment. A significant diversity of microorganisms in R. sanguineus females and in embryonic cell cultures from both lineages was found, with emphasis on the presence of Coxiella in all samples, albeit in different proportions. The Coxiella species present in the two lineages of ticks may be different and may have co-evolved with them, thus driving different patterns of interactions between ticks and the pathogens that they can harbor or transmit to vertebrate hosts.

scholarly journals THE PROLONGED GROWTH AND SURVIVAL OF OVARIAN TISSUE OF THE PROMETHEA MOTH (CALLOSAMIA PROMETHEA) IN VITRO

1958 ◽  
Vol 41 (5) ◽  
pp. 1027-1034 ◽  
Author(s):  
T. D. C. Grace
Keyword(s):  
Cell Culture ◽  
Cell Migration ◽  
Cell Cultures ◽  
Ovarian Tissue ◽  
Explant Culture ◽  
Growth And Survival ◽  
Live Tissue ◽  
Cell Divisions ◽  
Callosamia Promethea

1. The ovarian tissues from diapausing pupae of the promethea moth (Callosamia promethea) have survived and grown for 186 days under in vitro conditions. There was continual cell migration and multiplication for a period of 53 days, followed by a period of 47 days during which no cells migrated from the tissues. Between the 100th and 105th days after setting up the cultures, cell migration was resumed, and by the 111th day 250 cells were present in the medium. A few cell divisions were observed between the 126th and 136th days. After the tissues were subcultured on the 140th day, the explant culture continued to survive, but the cell culture died 3 days later. 2. The tissues were subcultured a total of 6 times during the 186 days. By the introduction of a piece of live tissue into the cell cultures, the growth and survival of the cells were increased from 8 days to about 20 days. 3. It is possible that the tissues had become adapted to the medium during their long survival, as the cells which migrated from them after 100 days showed considerably longer survival than those in earlier cultures.

Effects of Permeabilization on the Biotransformation of Phenylalanine by Immobilized Tobacco Cell Cultures

1989 ◽  
Vol 44 (3-4) ◽  
pp. 249-254 ◽  
Author(s):  
Jochen Berlin ◽  
Bernd Martin ◽  
Jerzy Nowak ◽  
Ludger Witte ◽  
Victor Wray ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Small Proportion ◽  
Culture Medium ◽  
Tobacco Cell ◽  
Permeabilized Cells ◽  
Oxidative Reactions ◽  
A Cell ◽  
Biotransformation Products ◽  
Severe Growth

Abstract Im mobilized tobacco cells excreted only a small proportion of their main secondary metabolite caffeoylputrescine into the culture medium . Experiments designed to release more of the com ­ pound by permeabilization caused the loss of caffeoylputrescine, probably by oxidative reactions. Moreover, rather mild treatments with permeabilizing agents (e.g. n-propanol) resulted in severe growth inhibition. The ability of permeabilized cells to form caffeoylputrescine and other hy-droxycinnam oyl conjugates from phenylalanine decreased considerably even when such cells were still able to metabolize phenylalanine into various ethyl acetate extractable com pounds (e.g. hydroxycinnamic acids and acetophenones). The formation of new biotransformation products suggests that permeabilized cells could be used as a tool for testing the enzymatic capabilities of a cell culture.

scholarly journals Cellular Effects of T-2 Toxin on Primary Hepatic Cell Culture Models of Chickens

Toxins ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 46 ◽  
Author(s):  
Máté Mackei ◽  
Kata Orbán ◽  
Andor Molnár ◽  
László Pál ◽  
Károly Dublecz ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Culture ◽  
Cell Cultures ◽  
Metabolic Activity ◽  
Incubation Time ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Cellular Effects ◽  
Culture Models ◽  
Cell Culture Models

Trichothecene mycotoxins such as T-2 toxin cause severe problems for agriculture, as well as for veterinary medicine. As liver is one of the key organs in metabolism, the main aim of our study was to investigate the immunomodulatory and cytotoxic effects of T-2 toxin, using primary hepatocyte mono-culture and hepatocyte—nonparenchymal cell (predominantly Kupffer cell) co-culture models of chicken. Cultures were exposed to 10 (T10 group), 100 (T100 group) and 1000 (T1000 group) nmol/L T-2 toxin treatment for 8 or 24 h. Alterations of cellular metabolic activity, the production of reactive oxygen species (extracellular H2O2), heat shock protein 70 (HSP70), and the concentration of different inflammatory cytokines such as interleukin (IL-)6 and IL-8 were investigated. Metabolic activity was intensely decreased by T-2 toxin administration in all of the cell culture models, in every applied concentration and incubation time. Concentrations of HSP70 and IL-8 were significantly increased in hepatocyte mono-cultures exposed to higher T-2 toxin levels (both in T100 and T1000 groups for HSP70 and in T1000 group for IL-8, respectively) compared to controls after 24 h incubation. Similarly, IL-6 levels were also significantly elevated in the T100 and T1000 groups in both of mono- and co-cultures, but only after 8 h of incubation time. In spite of the general harmful effects of T-2 toxin treatment, no significant differences were observed on reactive oxygen species production. Furthermore, the two cell culture models showed different levels of H2O2, HSP70, and IL-8 concentrations independently of T-2 toxin supplementation. In conclusion, the established primary cell cultures derived from chicken proved to be proper models to study the specific molecular effects caused by T-2 toxin. Metabolic activity and immune status of the different examined cell cultures were intensively affected; however, no changes were found in H2O2 levels.

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