Research Of High-Density Culture Of Porcine Circovirus Type2

In this paper, The research was PCV2was cultured and passaged on PK15cells,performed to optimize the main parameters of culturing PK15cells with microcarrier, produce cells and PCV2in high density using new bio-reactor technology. The experiments were conducted in three parts as follows:In experiment1, PCV2positive tissue homogenate was sterilized by filtratin and cultured on PK15cells without PCV contamination. The cultural character of PCV2was that D-glucosamine promoted the proliferation of PCV2effectively. But it could not be observed directly because no CPE was presented. PCR method was applied to detect the proliferation condition and titer of PCV2and to determine the harvest time of PCV2. The results showed that PCV2could proliferate and passage on PK15cells, and could be harvested duaring a certain period.In experiment2, some critical parameters were optimized including the influence of culture medium, density of cells inoculated and microcarrier concentration on the growth and metabolism of PK15cells. The results showed that the optimal cell density and microcarriver concentration were4.5X104cells/mg microcarrier and5-7mg/ml respectively. The optimal culture medium was DMEM-LG. The optimal culture condition was suitable for the production of viral vaccines.In experiment3, different culture systems for production PCV2were studied by using the bioreactor, roller bottle and square bottle. Then the titers of PCV2under different parameters were compared and optimized by using the roller bottle. The optimal culture conditions were that inoculating with1:10diluted virus-seed when the PK cells were grown for24hours,5-7mg/mL Micro-carrier concentration and low sugar DMEM containing0.2%LH as the maintenance fluid.