scholarly journals Permeability classification of representative fluoroquinolones by a cell culture method

AAPS PharmSci ◽  
2004 ◽  
Vol 6 (2) ◽  
pp. 1-6 ◽  
Author(s):  
Donna A. Volpe
Keyword(s):  
Cell Culture ◽  
Culture Method ◽  
A Cell

Automated cell counting of astrocytes on patterned substrates containing aliphatic and charged properties

1995 ◽  
Vol 53 ◽  
pp. 974-975
Author(s):  
W. Shain ◽  
H. Ancin ◽  
H.C. Craighead ◽  
M. Isaacson ◽  
L. Kam ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Attachment ◽  
Culture Method ◽  
Cell Counting ◽  
Data Sets ◽  
Nuclear Staining ◽  
Double Positive ◽  
A Cell ◽  
Wafer Test ◽  
Cell Densities

Neural protheses have potential to restore nervous system functions lost by trauma or disease. Nanofabrication extends this approach to implants for stimulating and recording from single or small groups of neurons in the spinal cord and brain; however, tissue compatibility is a major limitation to their practical application. We are using a cell culture method for quantitatively measuring cell attachment to surfaces designed for nanofabricated neural prostheses.Silicon wafer test surfaces composed of 50-μm bars separated by aliphatic regions were fabricated using methods similar to a procedure described by Kleinfeld et al. Test surfaces contained either a single or double positive charge/residue. Cyanine dyes (diIC18(3)) stained the background and cell membranes (Fig 1); however, identification of individual cells at higher densities was difficult (Fig 2). Nuclear staining with acriflavine allowed discrimination of individual cells and permitted automated counting of nuclei using 3-D data sets from the confocal microscope (Fig 3). For cell attachment assays, LRM5 5 astroglial cells and astrocytes in primary cell culture were plated at increasing cell densities on test substrates, incubated for 24 hr, fixed, stained, mounted on coverslips, and imaged with a 10x objective.

scholarly journals Tulane Virus Recognizes the A Type 3 and B Histo-Blood Group Antigens

2014 ◽  
Vol 89 (2) ◽  
pp. 1419-1427 ◽  
Author(s):  
Dongsheng Zhang ◽  
Pengwei Huang ◽  
Lu Zou ◽  
Todd L. Lowary ◽  
Ming Tan ◽  
...  
Keyword(s):  
Cell Culture ◽  
Blood Group ◽  
Culture Method ◽  
Blood Group Antigens ◽  
Content Type ◽  
Tulane Virus ◽  
A Cell ◽  
Type 3

ABSTRACTTulane virus (TV), the prototype of theRecovirusgenus in the calicivirus family, was isolated from the stools of rhesus monkeys and can be cultivatedin vitroin monkey kidney cells. TV is genetically closely related to the genusNorovirusand recognizes the histo-blood group antigens (HBGAs), similarly to human noroviruses (NoVs), making it a valuable surrogate for human NoVs. However, the precise structures of HBGAs recognized by TV remain elusive. In this study, we performed binding and blocking experiments on TV with extended HBGA types and showed that, while TV binds all four types (types 1 to 4) of the B antigens, it recognizes only the A type 3 antigen among four types of A antigens tested. The requirements for HBGAs in TV replication were demonstrated by blocking of TV replication in cell culture using the A type 3/4 and B saliva samples. Similar results were also observed in oligosaccharide-based blocking assays. Importantly, the previously reported, unexplained increase in TV replication by oligosaccharide in cell-based blocking assays has been clarified, which will facilitate the application of TV as a surrogate for human NoVs.IMPORTANCEOur understanding of the role of HBGAs in NoV infection has been significantly advanced in the past decade, but direct evidence for HBGAs as receptors for human NoVs remains lacking due to a lack of a cell culture method. TV recognizes HBGAs and can replicatein vitro, providing a valuable surrogate for human NoVs. However, TV binds to some but not all saliva samples from A-positive individuals, and an unexplained observation of synthetic oligosaccharide blocking of TV binding has been reported. These issues have been resolved in this study.

scholarly journals Thermal Stability of Hepatitis E Virus as Estimated by a Cell Culture Method

2016 ◽  
Vol 82 (14) ◽  
pp. 4225-4231 ◽  
Author(s):  
Reimar Johne ◽  
Eva Trojnar ◽  
Matthias Filter ◽  
Jörg Hofmann
Keyword(s):  
Cell Culture ◽  
Hepatitis E Virus ◽  
Culture Method ◽  
Hepatitis E ◽  
Pathogen Transmission ◽  
Heat Inactivation ◽  
Virus Inactivation ◽  
Genotype 3 ◽  
The Future ◽  
A Cell

ABSTRACTHepatitis E virus (HEV) is an increasingly recognized zoonotic pathogen. Transmission is suspected to occur from infected pigs or wild boars to humans through direct contact, environmental pathways, or contaminated food. However, the physical and chemical stability of HEV is largely unknown, because suitable cell culture methods for infectivity measurement are missing. Here, we developed a titration method using infection of the cell line A549/D3 with HEV genotype 3 strain 47832c and subsequent counting of focus-forming units by immunofluorescence, which allowed HEV infectivity measurements within a 4-log-dilution range. Long-term storage of HEV in cell culture medium at different temperatures indicated a phase of rapid virus inactivation, followed by a slower progression of virus inactivation. Infective HEV was detected up to 21 days at 37°C, up to 28 days at room temperature, and until the end of the experiment (56 days) with a 2.7-log decrease of infectious virus at 4°C. Heat treatment for 1 min resulted in moderate decreases of infectivity up to 60°C, 2- to 3.5-log decreases between 65°C and 75°C, and no remaining virus was detected at temperatures of ≥80°C. Heating for 70°C resulted in a 3.6-log decrease after 1.5 min and the absence of detectable virus (>3.9-log decrease) after 2 min. The data were used to calculate predictive heat inactivation models for HEV. The results may help estimate HEV stability in the environment or food. The established method may be used to study other aspects of HEV stability in the future.IMPORTANCEIn this study, a cell culture method was developed which allows the measurement of hepatitis E virus (HEV) infectivity. Using this system, the stability of HEV at different time-temperature combinations was assessed, and a predictive model was established. The obtained data may help estimate HEV stability in the environment or food, thus enabling an assessment of the relative risks of HEV infection through distinct routes and by distinct types of food in the future.

ASSESSMENT OF VIABILITY OF FROZEN BONE MARROW CELLS USING A CELL-CULTURE METHOD

1968 ◽  
Vol 1 (3) ◽  
pp. 255-261
Author(s):  
S. Abrahams ◽  
J. E. Till ◽  
E. A. McCulloch ◽  
L. Siminovitch
Keyword(s):  
Cell Culture ◽  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Culture Method ◽  
A Cell ◽  
Frozen Bone ◽  
Marrow Cells

scholarly journals Optimizing a Suspension Culture Method with a Decreased Cost to Detect Enteroviruses in Water to Increase Surveillance Access

2020 ◽  
Vol 11 (2) ◽  
pp. 35-44
Author(s):  
Stephanie Tornberg-Belanger ◽  
Jonathan Sreter ◽  
Aaron Margolin
Keyword(s):  
Cell Culture ◽  
Nucleic Acid ◽  
Water Samples ◽  
Financial Burden ◽  
Virus Detection ◽  
Culture Method ◽  
Virus Concentration ◽  
Viral Nucleic Acid ◽  
A Cell ◽  
Materials Used

Enteroviruses are a public health threat due to the high incidence of infections and potential for serious illness or death. Some laboratories in high-income countries detect enteroviruses in water by integrating cell culture and PCR (ICC/PCR). This combined method carries a high financial burden, due in part to specialized cell culture equipment. Therefore, we expanded upon a pilot study to reduce the cost by using common laboratory polypropylene tubes to create a cell culture in suspension. We optimized the protocol by determining minimal incubation periods post-infection as a function of the initial virus concentration. Cells in suspension and traditional monolayers were inoculated with poliovirus and incubated in 8-hour intervals up to 48 hours prior to extraction. Quantitative PCR (qPCR) was used to detect viral nucleic acid targets. Treated and raw water samples were seeded with virus and the suspension ICC/qPCR protocol used to ascertain whether the protocol performed similar to directly seeding cells. No variation in virus detection occurred using the suspension ICC/qPCR or monolayer ICC/qPCR (p = 0.95). In surface water samples, viral nucleic acid was successfully detected, with no significant increase after 32 h (p > 0.05). Suspension ICC/qPCR is as effective as monolayer ICC/qPCR in detecting enteroviruses in surface waters. Materials used in the suspension ICC/qPCR have a lower monetary cost than traditional cell culture materials without loss of sensitivity. More accessible testing of waters for enterovirus contamination through cost reduction has the potential to reduce human exposure and disease.

Comparison of Cell Culture and a Poliovirus Gene Probe Assay for the Detection of Enteroviruses in Environmental Water Samples

1993 ◽  
Vol 27 (3-4) ◽  
pp. 311-314 ◽  
Author(s):  
Aaron B. Margolin ◽  
Charles P. Gerba ◽  
Kenneth J. Richardson ◽  
Jaime E. Naranjo
Keyword(s):  
Cell Culture ◽  
Activated Sludge ◽  
Water Samples ◽  
Cdna Probe ◽  
Culture Method ◽  
Tidal River ◽  
Nucleic Acid Hybridization ◽  
Treated Wastewater ◽  
Gene Probe ◽  
Naturally Occurring

Nucleic acid hybridization provides a rapid non-cell culture method for the detection of enteric viruses in water. The purpose of this work was to compare the detection of naturally occurring enteroviruses by cell culture with their detection by a poliovirus gene probe in various types of water samples. Samples of activated sludge effluent, tertiary treated wastewater (activated sludge, filtration and passage through reverse osmosis), ground water, surface water and tidal river water were processed through 1 MDS Virozorb filters to concentrate any naturally occurring virus. Viruses were eluted from the filters with pH 9.5 beef extract and reduced in volume by flocculation to 20-30 ml. These concentrates were then assayed in the BGM cell line by the cytopathogenic effects (CPE) method and by a poliovirus cDNA probe (base pairs 115-7440) labeled with 32P. A total of 233 samples were assayed in this manner. In slightly more than 93% of the samples gene probe and cell culture yielded the same results. Of these samples 36 were positive by gene probe and 28 by cell culture assay. Positive samples for gene probe were confirmed by treatment with NaOH or RNAse and then reprobed. Samples demonstrating CPE upon primary passage were confirmed positive by subsequent passage of cell lysate on a new monolayer of BGM cells. Ten samples were positive by gene probe and negative by cell culture, and 4 samples were negative by gene probe and positive by cell culture.

Juglone Exerts Cytotoxic, Anti-proliferative and Anti-invasive Effects on Glioblastoma Multiforme in a Cell Culture Model

2016 ◽  
Vol 16 (9) ◽  
pp. 1190-1197 ◽  
Author(s):  
Dziugas Meskelevicius ◽  
Kastytis Sidlauskas ◽  
Ruta Bagdonaviciute ◽  
Julius Liobikas ◽  
Daiva Majiene
Keyword(s):  
Cell Culture ◽  
Glioblastoma Multiforme ◽  
Cell Culture Model ◽  
Culture Model ◽  
A Cell

A New In Vitro Model for Predicting the Toxicity of Cosmetic Products

1992 ◽  
Vol 20 (1) ◽  
pp. 138-143
Author(s):  
Maria Carrara ◽  
Lorenzo Cima ◽  
Roberto Cerini ◽  
Maurizio Dalle Carbonare
Keyword(s):  
Cell Culture ◽  
Cultured Cells ◽  
Neutral Red ◽  
Total Protein Content ◽  
Cosmetic Products ◽  
Cell Culture Insert ◽  
A Cell ◽  
Vitro Model ◽  
Cosmetic Emulsions

A method has been developed whereby cosmetic products which are not soluble in water or in alcohol can be brought into contact with cell cultures by being placed in a cell culture insert, which is then placed in the cell culture well. Preliminary experiments were carried out with L929 cells, and cytotoxicity was evaluated by measuring neutral red uptake and the total protein content of treated cultured cells. Encouraging results were obtained in comparisons of three cosmetic emulsions and of one emulsion containing a range of concentrations of two preservatives, Kathon CG and Bronopol.

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