2016
DOI: 10.1002/elsc.201600110
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H9 subtype influenza vaccine in MDCK single‐cell suspension culture with stable expression of TMPRSS2: Generation and efficacy evaluation

Abstract: Vaccination is the most effective way to protect chickens from avian influenza pandemics. Mammalian cells, especially MDCK cells, are being investigated as a good alternative to the embryonated chicken eggs for avian influenza vaccine production. In this work, we developed a new MDCK cell line, which has two main features: single‐cell suspension growth and stable expression of human TMPRSS2 protein (transmembrane protease serine S1 member 2) anchored on cells membrane, named MDCK‐Sus‐TMPRSS2. Reverse‐transcrip… Show more

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Cited by 4 publications
(2 citation statements)
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“…CHO cells with monolayer were transfected with the pCMV‐rE2 and different pTxnip‐rE2 vectors respectively using branched 25 kDa polyethylenimine (PEI, Sigma–Aldrich, 764604). The transfection procedure based on PEI was performed as previously described [14,15]. One day after the transfection, the different transfected cell groups were digested with trypsin and split into 10‐cm plates respectively in growth media with dialyzed FBS and without H/T addition for transgenic cells forming, followed with two rounds drug selection of 800 and 1200 μg/mL G418 (Roche, 04727878001).…”
Section: Methodsmentioning
confidence: 99%
“…CHO cells with monolayer were transfected with the pCMV‐rE2 and different pTxnip‐rE2 vectors respectively using branched 25 kDa polyethylenimine (PEI, Sigma–Aldrich, 764604). The transfection procedure based on PEI was performed as previously described [14,15]. One day after the transfection, the different transfected cell groups were digested with trypsin and split into 10‐cm plates respectively in growth media with dialyzed FBS and without H/T addition for transgenic cells forming, followed with two rounds drug selection of 800 and 1200 μg/mL G418 (Roche, 04727878001).…”
Section: Methodsmentioning
confidence: 99%
“…In this respect, it is a frequently preferred expression platform in influenza virus studies, vaccine development processes and vaccine production. ,, However, in order to improve the production process, a protease enzyme must be added to the production medium to convert hemagglutinin to its active form during the production process of influenza viruses . To eliminate this, there are studies conducted with MDCK cells modified to express TMPRSS2 (transmembrane protease, serine S1 family member 2) and HAT (human airway trypsin like protease) proteases, which altering hemagglutinin to the active form in natural influenza infection …”
Section: Animal Cell Culture-based Expression Platforms Used In Vacci...mentioning
confidence: 99%