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Iranian Journal of Biotechnology، جلد ۲۲، شماره ۲، صفحات ۷۴-۸۲
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عنوان انگلیسی Increased Expression Level of Human Blood Clotting Factor VIII Using NS0 Cell Line as a Host Cells
چکیده انگلیسی مقاله Background: Coagulation factor VIII (FVIII) is applied for spontaneous hemorrhaging inhibition and excessive bleeding 
after trauma in patients with hemophilia A. High-quality human recombinant factor VIII (rFVIII) has been produced 
relatively in large quantities in cultured mammalian cells. NS0 is one of the most common mammalian cell lines for 
therapeutic protein production. Production of rFVIII has increased due to low FVIII expression levels and rising demand 
for hemophilia A prophylactic treatment. Several methods have been developed to prevent cell cycle progression in 
mammalian cells for increased recombinant protein yields.
Objective: The aim of the study was to investigate the level of recombinant BDD-FVIII expression in NS0 mouse 
myeloma cells. Additionally, the study aimed to determine the effects of chemical drugs, Mitomycin C, Lovastatin, and 
Metformin on the secretion of FVIII through cell cycle arrest.
Materials and Methods: We cultured NS0 cells and transfected them with the 2 μg pcDNA3-hBDDFVIII plasmid by 
Lipofectamine 3000. The cells were treated with 10 μg.mL-1 Mitomycin C, 20 μM Lovastatin, and 20 mM Metformin 
separately. After 24 and 48 hours, the samples were collected and, protein expression was analyzed using RT-PCR, Dot 
blot, and ELISA.
Results: A higher protein expression level was observed in treated cells 24h and 48h after treatment with all three drugs. 
According to real-time PCR, Metformin treatment resulted in the highest expression level within 24 h (P=0.0026), followed by Mitomycin C treatment within 48 h (P=0.0030).
Conclusion: The NS0 cell line can be regarded as a suitable host for FVIII production. FVIII protein expression level was 
increased by using Lovastatin, Metformin, and Mitomycin C drugs. Further investigations are suggested, and the potential application of these drugs to increase recombinant protein yield can be used to produce therapeutic proteins in the industry.
کلیدواژه‌های انگلیسی مقاله Keywords Lovastatin,Metformin,Mitomycin C,NS0 mouse myeloma cells,recombinant factor VIII (rFVIII)
نویسندگان مقاله Mahboobeh Zarei |
Student Research Committee, Babol University of Medical Sciences, Babol, I.R. Iran.

Elaheh Ferdosi-Shahandashti |
Biomedical and Microbial Advanced Technologies (BMAT) Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, I.R. Iran.

Mohsen Badalzadeh |
Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran.

Gholam Ali Kardar |
Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran.

نشانی اینترنتی https://www.ijbiotech.com/article_196868_85fbe8bd37bf4a18d724d0a3de581c5f.pdf
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