Improvement of K562 Cell Line Transduction by Fbs Mediated Attachment to the Cell Culture Plate Publisher Pubmed



Abbasalipour M¹ ; Khosravi MA¹ ; Zeinali S¹ ; Khanahmad H² ; Karimipoor M¹ ; Azadmanesh K³ Show Affiliations
Source: BioMed Research International Published:2019

Abstract

Lentiviral vectors have been used for gene therapy in the clinical phase in recent years. These vectors provide a tool for gene insertion, deletion, or modification in organisms. The K562 human cell line has been used extensively in hematopoietic research. Despite its broad application, it is hard-to-transfection and transduction. So, this study presents a simple method to increase the transduction efficiency of K562 cells with a low multiplicity of infection (MOI) of the virus particle. For this purpose, 24-well plate was coated by 300 μl fetal bovine serum (FBS) before seeding. Then 2×10 4 K562 cells were seeded in each FBS coated plate. After 24h, K562 cells were attached and doubled. Different amount of lentivirus-based GFP vector according to MOI (5, 10, 15, and 20) along with 8 μg polybrene was added to the attached K562 cells and after 6h cells and viral particle complex were spinfected. Then cells were returned to the plate and incubated in 37°C overnight. After 48h transduction efficiency was established by measuring the GFP-expressing cells by flow cytometry. Flow cytometry analysis showed that, after plate treatment by FBS, 64.5% transduction rate in K562 cells was achieved at MOI=20. Therefore, this method can be an effective and simple way to increase the lentiviral transduction rate for suspended cells such as K562. © 2019 Maryam Abbasalipour et al.