Stress Factors Affecting the Cryopreservation of Biological Components of Seed Virus for Avian Influenza Vaccine Production

Muhammad Danish Mehmood, Muhammad Ahmad, Huma Anwar Ul-Haq, Muhammad Usman Ghani, Zahid Ali Tahir, Muhammad Ismail, Fareha Arshad, Abdul Rashid Shaukat
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Abstract

In the past, many studies have been done to cryopreserve biological materials for future vaccine production. Scientists have been using different chemicals as cryoprotectants to preserve their cell lines on which desired viruses can be cultivated. Researchers have always been in search for better molecules to avoid cryoinjury during process of cryopreservation. In the present study, different molecules were evaluated for cryo-potency in preserving master seeds of “Vero cell” line and “Avian Influenza viruses”. Cryoprotectants such as (Dimethyl sulfoxide) DMSO and Glycerol were used in different concentration and evaluated at -80oC and -196oC for different time interval. After 15 days and 30 days of cryopreservation the percentage viability of preserved cells were almost equal at both temperatures whereas, after 60 days, 90 days and 120 days the higher percentage of viability was recorded at -196oC for both Vero cells and Avian Influenza virus.  Different pH levels were set for both samples separately with same time interval and found slightly acidic pH (pH5.5) optimum for cryopreservation of Vero cell line and Neutral pH (pH7) for Avian Influenza virus. DMSO (20%) and Glycerol (40%) showed optimum percentage viability when cryopreserved for 120 days without any ill effect.  
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影响禽流感疫苗种子病毒生物成分低温保存的胁迫因素
在过去,已经进行了许多关于冷冻保存生物材料以用于未来疫苗生产的研究。科学家们一直在使用不同的化学物质作为冷冻保护剂来保存他们的细胞系,在这些细胞系上可以培养所需的病毒。在低温保存过程中,研究人员一直在寻找更好的分子来避免低温损伤。在本研究中,我们对不同分子在保存Vero细胞主种子中的低温效力进行了评价。禽流感病毒& &;ldquo;使用不同浓度的(二甲基亚砜)DMSO和甘油等冷冻保护剂,在-80℃和-196℃下进行不同时间间隔的评估。在低温保存15天和30天后,保存的细胞在两种温度下的存活率几乎相等,而在低温保存60天、90天和120天后,Vero细胞和禽流感病毒在-196℃下的存活率均较高。在相同的时间间隔内,对两种样品分别设置不同的pH值,发现微酸性pH值(pH5.5)最适合Vero细胞株的冷冻保存,中性pH值(pH7)最适合禽流感病毒的冷冻保存。DMSO(20%)和甘油(40%)冷冻保存120天,存活率最高,无不良反应。
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