| Highly pathogenic porcine reproductive and respiratory syndrome is an acute and highly lethal infectious disease caused by porcine reproductive and respiratory syndrome virus mutants,also known as highly pathogenic blue ear disease.The main clinical manifestations of sick pigs often showed a sudden rise in body temperature,high morbidity and high mortality.The epidemic situation outbreak in China since 2006.To the pig industry has brought a very heavy economic impact.The disease at this stage there is no special effects drugs,can only rely on vaccination for prevention.Most of our prevention of the use of attenuated vaccine,the immune effect is good,the advantages of the immune period is long.At present,the veterinary vaccines mainly bottle process,each batch difference is big,the quality is not stable.In order to obtain high-quality vaccine,the culture condition of Marc-145 cells was optimized and the cells were cultured in the suspension culture medium of bioreactor microcarriers and the culture parameters were further screened.The effects of cell culture medium pH,serum concentration and the proportion of subcultured cells on the culture of Marc-145 cells were optimized by single factor method.The results showed that the cell culture medium pH value was 7,cell culture medium serum concentration was 8%,cell passage ratio was 1: 3,Marc-145 cell culture was the best.In order to obtain a higher titer of porcine reproductive and respiratory syndrome virus(PRRSV)TJM strain on Marc-145 cells,the serum of the culture medium used for suspension culture of Marc-145 cells in bioreactor,culture medium,The amount of microcarriers and the time of virus culture were screened.The results showed that the cell culture time was 27 ~ 36 h,and the virus proliferation was in the range of 20 ~ 30 cells / ball and 5 g.Highest virus titer.The results of this study provide useful information for micro-vector production of PRRSV-TJM strain on a large scale. |
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