Application of Epstein–Barr Virus for Optimization of Immortalized B-lymphocyte Production as a Positive Control in Genetic Studies

Tousizadeh, Behnaz; Moghim, Sharareh; Chaleshtori, Ahmad Reza Salehi; Ghanbarian, Maryam; Mirian, Mina; Salehi, Mansoor; Tousizadeh, Sepideh; Zaboli, Fatemeh

Application of Epstein–Barr Virus for Optimization of Immortalized B-lymphocyte Production as a Positive Control in Genetic Studies

Document Type : Original Article

Authors

¹ Department of Microbiology, Faculty of Basic Sciences, Ayatollah Amoli Azad University, Amol; Pediatric Inherited Disease Research Center, Isfahan, Iran

² Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

³ Medical Genetics Center of Genome, Isfahan, Iran

⁴ Department of Microbiology, Faculty of Basic Science, Islamic Azad University, Falavarjan Branch, Isfahan, Iran

⁵ Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

⁶ Department of Microbiology, Faculty of Basic Sciences, Ayatollah Amoli Azad University, Amol; Medical Genetics Center of Genome, Isfahan; Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

⁷ Department of Health, School of Health, Shahrekord University of Medical Sciences, Shahrekord, Iran

⁸ Department of Microbiology, Faculty of Basic Sciences, Ayatollah Amoli Azad University, Amol, Iran

Abstract

Background: Infection of B-cells with Epstein–Barr virus (EBV) leads to more and subsequent immortalization. This is considered as the method of choice for generating lymphoblastoid cell lines (LCLs). Producing LCLs, although very useful but is very time consuming and troublesome, drives the requirement for quicker and more reliable methods for EBV-driven B-cell transformation.Materials and Methods: After successfully production of LCLs, different parameters including temperature, serum concentration, type of culture medium, and CO₂concentration were evaluated on EBV-transformed B-cells. In this study, we were able to produce LCLs and optimize condition. Results: The best condition for generating LCLs was 37°C, 5% CO₂, 20% fasting blood sugar, and RPMI 1640. The study results were to establish a reliable method for producing LCLs that can be used to produce immortalized B-cells from almost any sources. Conclusion: This can help with tumorgenecity studies, as well as producing control material for rare genetic disorders and so on. The aim of this study was to determine optimized condition for reliable and reproducible LCLs from different sources.

Keywords

References

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