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目的利用生物反应器-微载体培养技术培养Vero细胞制备柯萨奇病毒A组16型(Coxsackievirus A16,CA16)。方法应用生物反应器-微载体培养法进行Vero细胞培养,待细胞长成致密单层时,接种CA16,于37℃培养,每隔24 h观察细胞病变情况,并检测病毒滴度。结果 Vero细胞在微载体上吸附140 h后,绝大部分细胞在微载体上长成致密单层,细胞密度约为12.3×10~5个/ml;接种CA16后72 h,Vero细胞完全病变,几乎全部从微载体上脱落,病毒滴度达最高,约7.75 TCID_(50)/ml。结论成功采用生物反应器-微载体培养技术培养Vero细胞制备了CA16,且病毒滴度较高,为CA16灭活疫苗的研制奠定了基础。
Objective To prepare Coxsackievirus A16 (CA16) by culturing Vero cells with bioreactor - microcarrier technique. Methods Vero cells were cultured with bioreactor - microcarrier method. When the cells grew into a dense monolayer, CA16 cells were inoculated and cultured at 37 ℃. The cell lesions were observed every 24 hours and the virus titers were measured. Results Vero cells adsorbed on microcarriers for 140 h, most cells grew into dense monolayers on microcarriers with a cell density of about 12.3 × 10 ~ 5 / ml. Vero cells were completely diseased at 72 h after inoculation with CA16, Almost all shed from the microcarriers, the highest virus titer, about 7.75 TCID_ (50) / ml. Conclusion Successful cultivation of Vero cells using bioreactor-microcarrier culture technique resulted in the preparation of CA16 with high virus titer, which laid the foundation for the development of CA16 inactivated vaccine.