en عمومى General پژوهشي Original article <div><strong>Background and Aims:</strong> The&nbsp; ability&nbsp; of&nbsp; the&nbsp; reovirus&nbsp; to&nbsp; destroy&nbsp; various&nbsp; cancer&nbsp; cells&nbsp; in&nbsp; vitro&nbsp; and,&nbsp; the&nbsp; success&nbsp; of&nbsp; the&nbsp; virus&nbsp; in&nbsp; conducting&nbsp; clinical&nbsp; trial&nbsp; phases&nbsp; of &nbsp;cancer&nbsp; therapy&nbsp; has&nbsp; attracted&nbsp; the&nbsp; attention&nbsp; of&nbsp; researchers&nbsp; toward&nbsp; the&nbsp; virus.&nbsp; One&nbsp; of&nbsp; the&nbsp; main&nbsp; needs&nbsp; for&nbsp; the&nbsp; investigation&nbsp; of&nbsp; the&nbsp; viral&nbsp; effects&nbsp; and&nbsp; the&nbsp; virus-host&nbsp; cell&nbsp; interaction&nbsp; is&nbsp; preparation&nbsp; of&nbsp; the&nbsp; purified&nbsp; virus.&nbsp; Most&nbsp; of&nbsp; the&nbsp; protocols&nbsp; have&nbsp; been&nbsp; based&nbsp; on&nbsp; the&nbsp; use&nbsp; of&nbsp; suspended&nbsp; cell&nbsp; culture&nbsp; equipment&nbsp; that&nbsp; normally&nbsp; does&nbsp; not&nbsp; exist&nbsp; in&nbsp; the&nbsp; research&nbsp; laboratories.&nbsp; We&nbsp; optimized&nbsp; a&nbsp; virus&nbsp; purification&nbsp; method&nbsp; that&nbsp; was&nbsp; based&nbsp; on&nbsp; using&nbsp; cell&nbsp; culture&nbsp; flask&nbsp; and&nbsp; adherent&nbsp; cells<span dir="RTL">.</span><br> <strong>Materials and Methods:</strong> L-929 cells were used for reovirus propagation.&nbsp; After sufficient CPE, the cells were collected and pelleted.&nbsp; Using&nbsp; Vertrel-XF&nbsp; treatment&nbsp; and&nbsp; ultracentrifugation&nbsp; on&nbsp; the&nbsp; cesium&nbsp; chloride&nbsp; (CsCl)&nbsp; gradient,&nbsp; purified&nbsp; reovirus&nbsp; was&nbsp; obtained.&nbsp; It&nbsp; was&nbsp; subsequently&nbsp; concentrated&nbsp; by&nbsp; filtration&nbsp; using&nbsp; a&nbsp; 100kDa&nbsp; Amicon&nbsp; unit.&nbsp; Finally,&nbsp; infectivity&nbsp; and&nbsp; the&nbsp; number&nbsp; of&nbsp; purified&nbsp; human&nbsp; reoviruses&nbsp; were&nbsp; evaluated&nbsp; by&nbsp; plaque&nbsp; assay<span dir="RTL">.</span> The&nbsp; band&nbsp; of&nbsp; purified&nbsp; human&nbsp; reovirus&nbsp; was&nbsp; aspirated&nbsp; form&nbsp; the&nbsp; ultracentrifugation&nbsp; tube&nbsp; and&nbsp; then&nbsp; was&nbsp; dialysed&nbsp; and&nbsp; concentrated&nbsp; by&nbsp; filtration&nbsp; in&nbsp; Amicon&nbsp; unit.&nbsp; The&nbsp; titer&nbsp; of&nbsp; purified&nbsp; human&nbsp; reovirus&nbsp; was&nbsp; determined to be&nbsp; 3&times;1012&nbsp; PFUs/ml<span dir="RTL">.</span><br> <strong>Results:</strong> In&nbsp; present&nbsp; study,&nbsp; we&nbsp; established&nbsp; a&nbsp; protocol&nbsp; for&nbsp; the&nbsp; purification&nbsp; of&nbsp; human&nbsp; reovirus&nbsp; without&nbsp; need&nbsp; for&nbsp; equipment&nbsp; of&nbsp; suspension&nbsp; cell&nbsp; culture.&nbsp;<br> <strong>Conclusions: </strong>Although,&nbsp; the&nbsp; time-consuming&nbsp; dialysis&nbsp; procedure&nbsp; was&nbsp; removed&nbsp; from&nbsp; the&nbsp; end&nbsp; of&nbsp; the&nbsp; work&nbsp; and&nbsp; replaced&nbsp; with&nbsp; a&nbsp; rapid&nbsp; and&nbsp; simple&nbsp; filtration&nbsp; method,&nbsp; the&nbsp; high&nbsp; titer&nbsp; of&nbsp; purified&nbsp; human&nbsp; reovirus&nbsp; was&nbsp; acquired.&nbsp;&nbsp;&nbsp;</div> oncolytic virus, reovirus, virus purification, CsCl gradient. 35 40 http://journal.isv.org.ir/browse.php?a_code=A-10-234-1&slc_lang=en&sid=1 Mojtaba Hamidi-Fard mojtaba_hamidifard@yahoo.com 10031947532846007043 10031947532846007043 No Hepatitis & AIDS department, Pasteur institute of Iran, Tehran, Iran Angila Ataei-Pirkooh ataei.a@iums.ac.ir 10031947532846007044 10031947532846007044 No Department of Virology, Iran University of Medical Sciences, Tehran, Iran Mohammadreza Aghasadeghi mrasadeghi@pasteur.ac.ir 10031947532846007045 10031947532846007045 No Hepatitis & AIDS department, Pasteur institute of Iran, Tehran, Iran. Reihaneh Kazemi re.kazemi71@gmail.com 10031947532846007046 10031947532846007046 Yes Hepatitis & AIDS department, Pasteur institute of Iran, Tehran, Iran.