MDCK cell line with inducible allele B NS1 expression propagates delNS1 influenza virus to high titres

Vaccine ◽  
2011 ◽  
Vol 29 (40) ◽  
pp. 6976-6985 ◽  
Author(s):  
R. van Wielink ◽  
M.M. Harmsen ◽  
D.E. Martens ◽  
B.P.H. Peeters ◽  
R.H. Wijffels ◽  
...  
Vaccine ◽  
2013 ◽  
Vol 31 (48) ◽  
pp. 5693-5699 ◽  
Author(s):  
B. Peschel ◽  
S. Frentzel ◽  
T. Laske ◽  
Y. Genzel ◽  
U. Reichl

PLoS ONE ◽  
2013 ◽  
Vol 8 (9) ◽  
pp. e75014 ◽  
Author(s):  
Vladimir Y. Lugovtsev ◽  
Darya Melnyk ◽  
Jerry P. Weir

2020 ◽  
Author(s):  
Yixiao Wu ◽  
Hanjing Jia ◽  
Hanzhang Lai ◽  
Xuping Liu ◽  
Wen-Song Tan

Abstract H9N2 subtype avian influenza virus poses a constant threat to the poultry industry and the control of the disease leans upon the use of effective vaccines. As an alternative to the conventional chicken embryonated eggs, animal cell culture could overcome the limitations of egg supplies and upgrade the manufacturing of avian influenza vaccines for poultry. Development of serum-free suspension cell culture could allow even higher virus productivity, where a suspension cell line with good growth and production performance is required. In this work, an adherent MDCK cell line was adapted to suspension growth to cell concentration up to 12 × 106 cells/mL in a serum-free medium in batch cultures. Subsequently, the influenza virus propagation in this MDCK cell line was evaluated and was improved with the medium exchange at time of infection as well as optimization of infection conditions in terms of MOI and cell concentration for infection. Furthermore, various feed strategies were tested in the infection phase for improved virus titer and a maximum hemagglutinin titer of 13 log2 (HAU/50 μL) was obtained using the 1:2 medium dilution strategy. Evaluation of MDCK cell growth and H9N2 virus propagation in the bioreactors with optimized operating conditions showed comparable cell performance and virus yield compared to shake flasks, with a high cell-specific virus yield above 14000 virions/cell. With the purified H9N2 virus harvested from the bioreactor, the MDCK cell-derived vaccine was able to induce high titers of neutralizing antibodies in chickens. Overall, the results demonstrate the promising application of the highly efficient MDCK cell-based production platform for the avian influenza vaccine manufacturing.


2016 ◽  
Vol 106 ◽  
pp. 37-44 ◽  
Author(s):  
Yong Sun ◽  
Tingting Guo ◽  
Dawei Guo ◽  
Li Guo ◽  
Li Chen ◽  
...  

2018 ◽  
Vol 252 ◽  
pp. 94-99
Author(s):  
Viska I. Iskandar ◽  
Yutaka Sasaki ◽  
Naoto Yoshino ◽  
Raden Z.R. Abubakar ◽  
Shigehiro Sato ◽  
...  

1996 ◽  
Vol 15 (11) ◽  
pp. 904-908 ◽  
Author(s):  
Christopher D Lindsay

1 The epithelial Madin Darby Canine Kidney (MDCK) cell line was used to study the toxicity of ∈-toxin from Clostridium perfringens. The epithelial MDCK cell line is known to be sensitive to ∈-toxin of Clostridium perfringens and to investigate its mechanism of action, the neutral red assay has been used to determine the viability of cultures of this cell line. 2 Comparison of the LC 50s obtained at 34°C and 0°C showed that the lethality of ∈-toxin was reduced by 18- fold at the lower temperature. The effect of tempera ture on ∈-toxin lethality is unlikely to be due to reductions in membrane fluidity for the addition of Ca2+ or Mg2+ (2 mM) to buffer containing toxin was without effect. Varying the pH of the toxin-containing buffer from 6.9 to 8.7 did not increase the lethality of the toxin, though the most acidic pH used (5.8) was found to potentiate its action on MDCK cells. 3 The effect of inhibiting endocytosis on the lethality ofe toxin was also investigated by incubating cultures of MDCK cells with and without sodium azide over a range of concentrations of toxin. The co-administra tion of sodium azide did not reduce the toxicity of ∈ toxin, suggesting that energy-dependent uptake pro cesses such as endocytosis were unlikely to be involved in its mechanism of action. The results are, however, consistent with known receptor-based me chanisms of uptake and with other mechanisms of internalisation across the plasma membrane. ∈-toxin thus interacts with cell surfaces by a temperature sensitive mechanism potentiated by low pH.


Author(s):  
N. Mazurkova ◽  
E. Nechaeva ◽  
T. Ryabicheva ◽  
N. Varaksin ◽  
T. Sen'Kina ◽  
...  

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