微载体灌注培养Marc-145细胞制备猪繁殖与呼吸综合征疫苗  

Preparation of PRRS Vaccine by Perfusion Culture of Marc-145 Cells on Microcarriers

作　　者：杨雷 李伟 漆世华 秦红刚 韩兴 谢红玲 温文生 吴玉石 

机构地区：[1]武汉中博生物股份有限公司,武汉430070

出　　处：《中国兽药杂志》2013年第2期7-10,共4页Chinese Journal of Veterinary Drug

基　　金："十二五"农村领域国家科技计划课题"家畜病毒病基因工程疫苗创制"(2011AA10A208)

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。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 ×10^5/mL. After 4 days of culture, cell density can reach 5×10^6 - 7×10^6/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 1073TCID50/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.

关 键 词：微载体 灌注培养 猪繁殖与呼吸综合征疫苗 

分 类 号：S859.797[农业科学—临床兽医学]