摘要
应用激流式生物反应器培养Marc-145细胞生产高致病性猪繁殖与呼吸综合征病毒,并通过工艺优化,实现了病毒抗原的高效生产。首先将Marc-145细胞用含6 0mL/L牛血清的DMEM培养液复苏放大培养,当细胞量达到3×109时,接种入反应器中。先用细胞生长液培养,当细胞达到最大量时更换生长液为维持液,并接种HP-PRRSV。整个过程采用流加方式,每8h采样测定培养上清中葡萄糖浓度。接种病毒后,每24h测定培养上清HP-PRRSV滴度。6个批次细胞生长至88h,糖耗达到最高水平。连续3个批次种毒后培养至96h,上清中HP-PRRSV滴度达到最高,平均约为每106.4 TCID50/0.1mL。因此,认为应用激流式生物反应器进行细胞培养,通过过程工艺优化,可以实现HP-PRRSV抗原的高滴度生产。
In this study,the current bioreactor was used for the culture of African green monkey kidney cells（Marc-145） to achieve high pathogenic porcine reproductive and respiratory syndrome virus（HP-PRRSV） antigen in large scale and high titer production.Firstly,Marc-145 cell strain was first cultured in flasks with DMEM medium containing 6 percent fetal bovine serum（FBS）.When the cells reached about 3×109,they were seeded into the current bioreactor.The medium containing 6 percent FBS was used for the culture of Marc-145,and then it was replaced by the medium containing 2 percent FBS and HP-PRRSV when the cells reached the maximum number.Fed-batch culture was performed in the process of cell growth and virus proliferation.Sample was collected once every 8 hours for the measurement of glucose concentration throughout the fed-batch culture.After the virus was inoculated,the HP-PRRSV titer of culture supernatant was measured every 24 h.The six batches of cells were grown to 88h,and reached the highest level of glucose consumption.When three consecutive batches of cells cultured 96 h after virus inoculation,the highest average titer of supernatant HP-PRRSV was approximately 0.1 mL 106.4 TCID50.Therefor,through the optimization of process,the efficient production of HP-PRRSV antigen was achieved in the process of application current bioreactor to cultivate Marc-145 cells.
出处
《动物医学进展》
CSCD
北大核心
2012年第11期80-84,共5页
Progress In Veterinary Medicine
关键词
生物反应器
细胞培养
高致病性猪繁殖与呼吸综合征病毒
bioreactor
cell culture
highly pathogenic porcine reproductive and respiratory syndrome virus
