scholarly journals Bovine endometrium-derived cultured cells are suitable for lipofection

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mai Shiokawa ◽  
Ryotaro Miura ◽  
Aki Okubo ◽  
Yujiro Hagita ◽  
Itaru Yoshimura ◽  
...  

AbstractBovine-derived cultured cells, including Madin-Darby bovine kidney cells, are used worldwide; however, lipofection tend to result in low transfection efficiency, which has impeded the progress of veterinary research. We performed experiments to confirm the lipofection efficiency of bovine-derived cultured cells, to identify cells that suitable for lipofection. Several bovine tissues (endometrium, testis, ear tissue and foetal muscle) were collected, and primary cultured cells were prepared. Lipofection assay showed that only bovine endometrium (BE)-derived cells could be transfected efficiently (50‒70%). BE cells can be divided into at least two types of cell populations (BE-1 and BE-2). The BE-1 cells, which were suitable for lipofection, were obtained by passages at short intervals and were negative for cytokeratin- and positive for vimentin-expression; the BE-2 cells did not have these characteristics and were not suitable for lipofection. Furthermore, the BE-1 cells and artificially immortalised cells of BE-1, iBE-1 cells, were utilised in a reporter assay requiring the introduction of multiple DNAs. Endometrial tissues can be collected from living cows, and BE-1 cells can be obtained easily by controlling passaging timing. The production of BE-1 cells and sharing the methods required to prepare them will contribute to the development of veterinary research.

1984 ◽  
Vol 224 (3) ◽  
pp. 1001-1008 ◽  
Author(s):  
A R P Paterson ◽  
E R Harley ◽  
C E Cass

Dilazep, a vasodilator previously recognized as an inhibitor of adenosine permeation, very rapidly blocked the uptake of adenosine by cultured L5178Y cells, and accordingly was used as a quencher in a simple quenched-flow system for measuring cellular uptake of nucleosides during very short intervals. Time courses of cellular uptake of adenosine, assayed during intervals between 0.05 and 0.5s with the quenched-flow system, were linear and defined initial rates of adenosine uptake. The latter are rates of inward transport of adenosine. Kinetic constants for that process in cultured S49 cells determined with the quenched-flow procedure were similar to those determined with an assay dependent on manual timing. In studies of adenosine uptake kinetics in human erythrocytes at 22 degrees C and 37 degrees C in which the quenched-flow procedure was used, time courses of adenosine uptake were linear at both temperatures and defined initial uptake rates; kinetic constants (means +/- S.E.M.) at 22 degrees C (n = 8) were Km 25 +/- 14 microM and Vmax. 15 +/- 5 pmol/s per microliter of cell water and at 37 degrees C (n = 3) were Km 98 +/- 17 microM and Vmax. 80 +/- 9 pmol/s per microliter of cell water.


1979 ◽  
Vol 9 (2) ◽  
pp. 269-273
Author(s):  
M Fishaut ◽  
K McIntosh ◽  
G Meiklejohn

During the winter of 1977-1978 three influenza A virus serotypes (A/Vic/3/75, A/Texas/1/77 [both H3N2], and A/USSR/90/77 [H1N1]) circulated in Denver, offering us the opportunity to apply fluorescent antibody techniques to the specific identification of these viruses. Surface antigens of infected, unfixed primary monkey kidney cells were stained in suspension by an indirect immunofluorescence technique with anti-H3N2 and anti-H1N1 antisera. In tests of cells infected with known viruses, the members of the H3N2 family could not be distinguished from one another, but were easily distinguished from H1N1 strains. A total of 101 hemadsorption-positive clinical specimens were evaluated over a 6-month period. Forty-five of 48 influenza A H3N2 and 24 of 29 H1N1 specimens confirmed by hemagglutination inhibition were correctly identified by membrane fluorescence of cultured cells, with no misidentifications among influenza strains and with 1 false positive among 24 non-influenza isolates. The average time to identification by this technique was 4 days compared to 7 days by hemagglutination inhibition. Live cell membrane fluorescence is a simple, rapid, and accurate method for identifying and grouping influenza A viruses.


2020 ◽  
Vol 241 ◽  
pp. 108573 ◽  
Author(s):  
Jizong Li ◽  
Li Mao ◽  
Fang Xiao ◽  
Zheng Liao ◽  
Junlei Yin ◽  
...  

Planta Medica ◽  
1985 ◽  
Vol 51 (03) ◽  
pp. 194-197 ◽  
Author(s):  
T. Narasimhan ◽  
N. Harindranath ◽  
S. Premlata ◽  
B. Murthy ◽  
P. Rao

2011 ◽  
Vol 92 (6) ◽  
pp. 1398-1409 ◽  
Author(s):  
Axel Mauroy ◽  
Laurent Gillet ◽  
Elisabeth Mathijs ◽  
Alain Vanderplasschen ◽  
Etienne Thiry

Bovine noroviruses belong to the family Caliciviridae, genus Norovirus. Two genotypes have been described and viruses genetically related to the Jena and Newbury2 strains have been classified into genotypes 1 and 2, respectively. In this study, virus-like particles (VLP) of the previously detected B309 Belgian strain, genetically related to genotype 2 bovine noroviruses, were used to investigate virus–host interactions in vitro. B309 VLP were shown to bind to several bovine cell lines. This binding was not affected by heparinase or chondroitinase treatment but was significantly inhibited by both sodium periodate, α-galactosidase, trypsin and phospholipase C treatment. Cell treatment by neuraminidase also moderately affected this binding. Taken together, these results show that, in addition to a galactosyl residue, sialic acid could also be involved in binding to susceptible cells. In addition, both the cholesterol-dependent pathway and macropinocytosis are used for B309 VLP internalization by Madin–Darby bovine kidney cells. The data increase the knowledge on bovine norovirus cell interactions.


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