Hepatitis & AIDS department, Pasteur institute of Iran, Tehran, Iran.
Abstract: (2127 Views)
Background and Aims: The ability of the reovirus to destroy various cancer cells in vitro and, the success of the virus in conducting clinical trial phases of cancer therapy has attracted the attention of researchers toward the virus. One of the main needs for the investigation of the viral effects and the virus-host cell interaction is preparation of the purified virus. Most of the protocols have been based on the use of suspended cell culture equipment that normally does not exist in the research laboratories. We optimized a virus purification method that was based on using cell culture flask and adherent cells. Materials and Methods: L-929 cells were used for reovirus propagation. After sufficient CPE, the cells were collected and pelleted. Using Vertrel-XF treatment and ultracentrifugation on the cesium chloride (CsCl) gradient, purified reovirus was obtained. It was subsequently concentrated by filtration using a 100kDa Amicon unit. Finally, infectivity and the number of purified human reoviruses were evaluated by plaque assay. The band of purified human reovirus was aspirated form the ultracentrifugation tube and then was dialysed and concentrated by filtration in Amicon unit. The titer of purified human reovirus was determined to be 3×1012 PFUs/ml. Results: In present study, we established a protocol for the purification of human reovirus without need for equipment of suspension cell culture. Conclusions: Although, the time-consuming dialysis procedure was removed from the end of the work and replaced with a rapid and simple filtration method, the high titer of purified human reovirus was acquired.
Hamidi-Fard M, Ataei-Pirkooh A, Aghasadeghi M, Kazemi R. Purification of Human Reovirus in Monolayer of L-929 Cells. Iran J Virol 2019; 13 (1) :35-40 URL: http://journal.isv.org.ir/article-1-368-en.html