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当前位置: 首页 > 文章 > 微载体灌注培养Marc-145细胞制备猪繁殖与呼吸综合征疫苗 中国兽药杂志 2013,47 (2) 7-10
Position: Home > Articles > Preparation of PRRS Vaccine by Perfusion Culture of Marc-145 Cells on Microcarriers Chinese Journal of Veterinary Drug 2013,47 (2) 7-10

微载体灌注培养Marc-145细胞制备猪繁殖与呼吸综合征疫苗

作  者：
杨雷;李伟;漆世华;秦红刚;韩兴;谢红玲;温文生;吴玉石
单  位：
武汉中博生物股份有限公司
关键词：
微载体;灌注培养;猪繁殖与呼吸综合征疫苗
摘  要：
对在生物反应器中用微载体连续灌注培养Marc-145细胞生产猪繁殖与呼吸综合征病毒的制备技术进行了研究。在14 L体积的生物反应器中,加入含10 g/L微载体的细胞培养基DMEM,接种Marc-145细胞至细胞浓度为1×105/mL,培养4 d后细胞可生长至5～7×106/mL,然后以感染复数(MOI)为0.01接种PRRSV PC株病毒,接毒后36 h开始收获,连续收获3 d左右,收获的病毒滴度范围在106.0～107.3TCID50/mL之间,将收获的病毒液加入适量的保护剂,经冷冻干燥制备成疫苗,无菌、支原体等项目的检验均合格,3批疫苗的免疫保护率均为5/5。实验表明,用生物反应器微载体灌注培养Marc-145细胞制备PRRS疫苗工艺可行。
译  名：
Preparation of PRRS Vaccine by Perfusion Culture of Marc-145 Cells on Microcarriers
作  者：
YANG Lei,LI Wei,QI Shi-hua,QIN Hong-gang,HAN Xing,XIE Hong-ling,WEN Wen-sheng,WU Yu-shi(Wuhan Chopper Biology Co.,Ltd,Wuhan 430070,China)
关键词：
microcarrier;perfusion culture;PRRS vaccine
摘  要：
A technique for preparing porcine reproductive and respiratory syndrome(PRRS) vaccine by perfusion culture of Marc-145 cells on microcarriers in bioreactors was established.A 14-liter bioreactor was filled with DMEM cell culture medium which contained 10 g/L of microcarriers,Marc-145 cells were inoculated with a final cell density of 1×105/mL.After 4 days of culture,cell density can reach 5×106~7×106/mL.Then PRRSV PC strain was inoculated with a MOI of 0.01.After inoculation for 36 hours,the cultured cells were harvested continuously for 3 days.The titer of virus in the culture medium was about 106.0 to 107.3TCID50/mL,which was then added with suitable amount of protective agent and freeze-dried to form vaccine.The aseptic,Mycoplasma and other tests were used to qualify.The immune protective rate of the three batches vaccine was 5/5,which demonstrated that the preparation technology of PRRS vaccine by perfusion culture of Marc-145 cells on microcarriers in bioreactors was viable.

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