fa ﻣﻘﺎﯾﺴﻪ رده ﺳﻠﻮل ﻫﺎی A549‬ و L929 ﺑﺮای ﺗﮑﺜﯿﺮ وﯾﺮوس ‪EMCV ویروس شناسی پزشکی Medical Virology مقاله پژوهشی Original Research Article <div align="right" style="direction: rtl"> <b>زﻣﯿﻨﻪ وﻫﺪف:</b> وﯾﺮوس ‪  EMCV‬از ﺧﺎﻧﻮاده ﭘﯿﮑﻮرﻧﺎوﯾﺮﯾﺪه و از ﺟﻨﺲ ﮐﺎردﯾﻮوﯾﺮوس ﻫﺎ ﺑﻮده واز ﻧﺘﺎﯾﺞ ﺗﮑﺜﯿﺮ اﯾﻦ وﯾﺮوس‬ ‫در رده ﻫﺎﯾﯽ ﻧﻈﯿﺮ A549‬ ، L929 ‬در ﻣﻄﺎﻟﻌﺎﺗﯽ ﻧﻈﯿﺮ ﺳﻨﺠﺶ ﻣﻘﺎدﯾﺮ اﯾﻨﺘﺮﻓﺮون ﻫﺎ وﺗﻌﯿﯿﻦ ﻓﻌﺎﻟﯿﺖ زﯾﺴﺘﯽ آﻧﻬﺎ وﺗﺤﻘﯿﻘﺎت‬ ‫ﻣﺘﻌﺪد دﯾﮕﺮ ﺑﻪ ﮐﺜﺮت ﺑﻪ ﮐﺎر ﮔﺮﻓﺘﻪ ﺷﺪه اﺳﺖ. اﯾﻦ ﺗﺤﻘﯿﻖ ﺑﺎ ﻫﺪف ﺑﺮرﺳﯽ ﻣﻘﺎﯾﺴﻪ ای ﺗﮑﺜﯿﺮ وﯾﺮوس ‪ EMCV‬ در دو رده‬ ‫ﺳﻠﻮﻟﯽ A549‬ ، L929‬ ﺑﺮای ﺗﻌﯿﯿﻦ ﭘﺎﯾﻪ ﺳﻠﻮﻟﯽ ﻣﻨﺎﺳﺐ ﺗﺮ ﺑﺮای ﺗﮑﺜﯿﺮ وﯾﺮوس‪ EMCV‬اﻧﺠﺎم ﺷﺪ.‬ <br>‫<b>روش ﺑﺮرﺳﯽ:</b> در اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﭘﺲ از ﺗﻌﯿﯿﻦ و ﻣﻘﺎﯾﺴﻪ ﮐﯿﻨﺘﯿﮏ رﺷﺪ دو رده ﺳﻠﻮل A549‬ ، L929 ‬ﺑﻪ ﻣﺪت 5 ﺷﺒﺎﻧﻪ روز‬ ‫ﺗﺤﺖ ﺷﺮاﯾﻂ ﯾﮑﺴﺎن، ﺗﺎﺛﯿﺮ ﻧﻮع رده ﺳﻠﻮﻟﯽ در ﺗﮑﺜﯿﺮ وﯾﺮوس ‪  EMC‬ﺑﺎ ﺗﻌﯿﯿﻦ ﺗﯿﺘﺮ وﯾﺮوس  (CCID50) ﭘﺲ از ﮔﺬﺷﺖ 5‬ ‫ﺷﺒﺎﻧﻪ روز در ﻫﺮ رده ﺳﻠﻮﻟﯽ  A549‬ ، L929 ‬ﺑﺎ ﺗﻠﻘﯿﺢ وﯾﺮوس از ﯾﮏ اﺳﺘﻮک ﻣﺸﺎﺑﻪ وﯾﺮوﺳﯽ ﺑﺎ ﻏﻠﻈﺖ ﻫﺎی ﻣﺨﺘﻠﻒ در‬ ‫ﻫﺮ رده ﺑﻪ ﺻﻮرت ﻣﺠﺰا ﺑﺎ روش ‪  Reed-Muench‬وﻧﻬﺎﯾﺘﺎً رﻧﮓ آﻣﯿﺰی ‪  MTT‬و ﺳﻨﺠﺶ ‪  OD‬ﺗﻮﺳﻂ دﺳﺘﮕﺎه اﻻﯾﺰا رﯾﺪر‬ ‫اﻧﺠﺎم ﮔﺮﻓﺖ.‬ ‫<br><b>ﯾﺎﻓﺘﻪ ﻫﺎ:</b> ﺗﯿﺘﺮ وﯾﺮوس ﺑﻪ دﺳﺖ آﻣﺪه ﺑﻪ روش ‪ ,Reed_Muench‬ در رده ﺳﻠﻮل A549 در 24 ﺳﺎﻋﺖ اول ﭘﺲ از ﺗﻠﻘﯿﺢ‬ ‫وﯾﺮوس ﺑﻪ ﺳﻠﻮل از ﯾﮏ اﺳﺘﻮک ﻣﺸﺎﺑﻪ،  ^1.5CCID50 / ml,‬ 10 و درﻣﻘﺎﺑﻞ در رده ﺳﻠﻮل L929 ‬اﯾﻦ ﻣﻘﺪار ﺑﻪ 10^3.5CCID50 / ml اﻓﺰاﯾﺶ ﯾﺎﻓﺖ و ﺑﻪ ﻫﻤﯿﻦ ﺗﺮﺗﯿﺐ در روز ﻫﺎی ﺑﻌﺪی ﻣﻘﺎدﯾﺮ ﺗﯿﺘﺮ وﯾﺮوس ﺑﯿﺸﺘﺮی در رده L929 ‬ﮔﺰارش ﺷﺪ. ﺗﯿﺘﺮ‬ ‫وﯾﺮوﺳﯽ در روز ﺳﻮم در رده A549‬ در ﻣﻘﺪار  ﻣﺘﻮﻗﻒ در ﺻﻮرﺗﯽ ﮐﻪ ﺗﮑﺜﯿﺮ وﯾﺮوس در رده 929‪L‬‬ ‫ﺗﺎ روز ﭼﻬﺎرم اداﻣﻪ داﺷﺖ وﭘﺲ از آن در ﻣﻘﺪار 5/6 01 ‪ CCID50/ ml‬ﺛﺎﺑﺖ ﺷﺪ وﭘﺲ ازﻣﻄﺎﻟﻌﻪ ﻣﯿﮑﺮوﺳﮑﻮﭘﯽ ‪ ، CPE‬رﻧﮓ‬ ‫آﻣﯿﺰی ‪ MTT‬اﻧﺠﺎم ﺷﺪ و ﺗﯿﺘﺮ وﯾﺮوﺳﯽ 10^3.5CCID50 /  ml ﺑﺮای رده A945و 10^1.5CCID50 / ml برای رده L929‬‬ ‫ﮔﺰارش ﺷﺪ، ﮐﻪ ﻣﺸﺎﺑﻬﺖ زﯾﺎدی ﺑﯿﻦ ﺗﯿﺘﺮ وﯾﺮوﺳﯽ ﻣﺤﺎﺳﺒﻪ ﺷﺪه از ﻫﺮ دو روش دﯾﺪه ﺷﺪ.‬ ‫<br>ﻧﺘﯿﺠﻪ ﮔﯿﺮی: وﯾﺮوس ‪  EMCV‬ﻗﺎﺑﻠﯿﺖ رﺷﺪ در دو رده A549‬ و L929  ‬را دارا ﻣﯽ ﺑﺎﺷﺪ. رده ﺳﻠﻮﻟﯽ L929 ﮐﯿﻨﺘﯿﮏ‬ ‫رﺷﺪ ﺳﺮﯾﻊ ﺗﺮی ﻧﺴﺒﺖ ﺑﻪ رده ﺳﻠﻮﻟﯽ A945 ‬داراﺳﺖ. رده ﺳﻠﻮﻟﯽL929 ‬ﺑﺮای ﺗﮑﺜﯿﺮ وﯾﺮوس ‪  EMCV‬ﻧﺴﺒﺖ ﺑﻪ رده‬ ‫A549 ‬از ﻗﺎﺑﻠﯿﺖ ﺑﯿﺸﺘﺮی ﺑﺮﺧﻮردار اﺳﺖ و ﺗﯿﺘﺮ وﯾﺮوﺳﯽ ﺑﺎﻻﺗﺮی را ﻧﺸﺎن ﻣﯽ دﻫﺪ و ﺑﺮای ﺗﮑﺜﯿﺮ وﯾﺮوس ‪EMCV‬‬ ‫ﻣﻨﺎﺳﺐ ﺗﺮ اﺳﺖ اﯾﻦ اﻣﺮ ﺑﺮای ﺟﻠﻮﮔﯿﺮی از ﻫﺪر رﻓﺖ ﻫﺰﯾﻨﻪ و زﻣﺎن و اﻧﺮژی در ﻣﻄﺎﻟﻌﺎت واﺑﺴﺘﻪ ﺑﻪ ﺗﮑﺜﯿﺮ اﯾﻦ وﯾﺮوس ﻧﻈﯿﺮ‬ ‫ﺗﻌﯿﯿﻦ ﻋﯿﺎر ﻣوﻠﮑﻮﻟ های داروﯾﯽ ﻣﻬﻢ در درﻣﺎن ﺑﯿﻤﺎرﯾﻬﺎ ﺿﺮوری ﺑﻪ ﻧﻈﺮ ﻣﯽ رﺳﺪ ﺷﺎﯾﺎن ذﮐﺮ اﺳﺖ ﮐﻪ ﺗﺎﮐﻨﻮن ﻣﻄﺎﻟﻌﻪ ﻣﺸﺎﺑﻬﯽ‬ ‫در اﯾﻦ زﻣﯿﻨﻪ ﺻﻮرت ﻧﮕﺮﻓﺘﻪ اﺳﺖ.‬ </div> <b>Background: </b>EMCV virus belongs to the Picornaviride family and Cardioviruses genus. Replication results of EMCV is applicable for L929 and A549 cell lines on studies related to measure the potency of proteins such as interferon and biological assessment and other various researches. In order to choose the appropriate virus cell line, comparison of the EMCV virus replication in A549 and L929 cell lines is experimentally investigated. The aim of the present study is to compare the kinetic of growth of EMCV in A549and L929cell lines. <br><b>Material and method:</b> The goal of this study is to investigate the effect of the EMCV virus with the determination of the virus titer (CCID50) after passing five days on A549 and L929 cell lines under same condition. After a specific time (5 days), these cell lines are cultivated with the same virus in different densities with Reed-Munch method. MTT coloring and OD assessment are carried out with the Elayza reader system. <br><b>Result: </b>The A549 virus titer obtained from Reed-Munch method after 24 hours reached 10^{1.5 CCID50}/ ml, while the virus titer of 10 ^{3.5 CCID50}/ml is obtained for L929 virus. In the following days the more virus titer is reported for L929 virus. A549 virus titer stopped after 3days with the quantity of 10 ^{3.5 CCID50} /ml while the L929 cell line titer continued to grow for the next four days and reached 10^{6.5 CCID50}/ml. . MTT coloring was done after CPE study and virus titers of 10 ^{3.5 CCID50}/ml and 10 ^{6.5 CCID50}/ml are reported for the A549 and L929 respectively. <br><b>Conclusion: </b>Although EMCV virus has an ability to grow in A549 and L929 cell lines, the growth kinetic of the L929is faster than A549. Showing the higher virus titer, it can be concluded that L929is more appropriate for the EMCV cultivation. In studies related to EMCV, Using L929cell line in studies causes less time and cost to be consumed. آﻧﺴﻔﺎﻟﻮﻣﯿﻮﮐﺎردﯾﺘﯿﺲ، وﯾﺮوسEMCV، ‬اﺛﺮات ﺳﺎﯾﺘﻮﭘﺎﺗﯿﮏ‬، A549‬ ، L929 Encephalomyocarditis virus, A549, L929, Cytopatic effect 29 37 http://ijmm.ir/browse.php?a_code=A-10-1-157&slc_lang=fa&sid=1 Hoda Yaghmaiean ﻫﺪی ﯾﻐﻤﺎﺋﯿﺎن hoda.yaghmaiean@yahoo.com 1003194753284600949 1003194753284600949 No Faculty of Basic Science , Azad University داﻧﺸﮑﺪه ﻋﻠﻮم ﭘﺎﯾﻪ، داﻧﺸﮕﺎه آزاد اﺳﻼﻣﯽ واﺣﺪ ﻋﻠﻮم ﺗﺤﻘﯿﻘﺎت Fazeli ﻣﺤﻤﺪ رﺿﺎ ﻓﺎﺿﻠﯽ fazelimoSinatums@yahoo.com 1003194753284600950 1003194753284600950 Yes department of drug and food control, Pharmacy Faculty, Medical University of Tehran داﻧﺸﮑﺪه داروﺳﺎزی داﻧﺸﮕﺎه ﻋﻠﻮم ﭘﺰﺷﮑﯽ ﺗﻬﺮان‬ Jamlifar ﺣﺴﯿﻦ ﺟﻤﺎﻟﯽ فر 1003194753284600951 1003194753284600951 No department of drug and food control, Pharmacy Faculty, Medical University of Tehran ﺷﺮﮐﺖ زﯾﺴﺖ دارو، ﻣﺮﮐﺰ رﺷﺪ وﻓﻨﺎوری ﻓﺮاورده ﻫﺎی داروﯾﯽ Ilka Hoshmand ایلکا هوشمند 1003194753284600952 1003194753284600952 No R&D department of Zist-Daro company ﮔﺮوه ﮐﻨﺘﺮل دارو و ﻏﺬا داﻧﺸﮑﺪه داروﺳﺎزی داﻧﺸﮕﺎه ﻋﻠﻮم ﭘﺰﺷﮑﯽ ﺗﻬﺮان Mohhamad Taghavian محمد تقویان 1003194753284600953 1003194753284600953 No department of drug and food control, Pharmacy Faculty, Medical University of Tehran department of drug and food control, Pharmacy Faculty, Medical University of Tehran