BioReliance Corporation, 9900 Blackwell Road, Rockville, Maryland, 20850, USA.
Cytotechnology. 1999 Jul;30(1-3):169-72. doi: 10.1023/A:1008040221630.
We have compared three different production methods, which may be suitable for the large scale production of adenovirus vectors for human clinical trials. The procedures compared 293 cells adapted to suspension growth in serum-free medium in a stirred tank bioreactor, 293 cells on microcarriers in serum-containing medium in a stirred tank bioreactor, and 293 cells grown in standard tissue culture plasticware. With a given virus, yields varied between 2000 and 10,000 infectious units/cell. The stirred tank bioreactor routinely produced between 4000 and 7000 infectious units/cell when 293 cells were grown on microcarriers. The 293 cells adapted to suspension growth in serum-free medium in the same stirred tank bioreactor yielded between 2000 and 7000 infectious units/cell. Yields obtained from standard tissue culture plasticware were up to 10,000 infectious units/cell. Cell culture conditions were monitored for glucose consumption, lactate production, and ammonia accumulation. Glucose consumption and lactate accumulation correlated well with the cell growth parameters. Ammonia production does not appear to be significant. Based on virus yields, ease of operation and linear scalability, large-scale adenovirus production seems feasible using 293 cells (adapted to suspension/serum free medium or on microcarriers in serum containing medium) in a stirred tank bioreactor.
我们比较了三种不同的生产方法，这些方法可能适用于大规模生产用于人体临床试验的腺病毒载体。所比较的程序包括在搅拌罐生物反应器中悬浮生长于无血清培养基中的适应悬浮生长的 293 细胞、在含血清培养基中的微载体上的 293 细胞，以及在标准组织培养塑料器皿中生长的 293 细胞。对于给定的病毒，产率在 2000 到 10000 感染单位/细胞之间变化。当 293 细胞在微载体上生长时，搅拌罐生物反应器通常产生 4000 到 7000 感染单位/细胞。在相同的搅拌罐生物反应器中适应悬浮生长于无血清培养基中的 293 细胞产生 2000 到 7000 感染单位/细胞。从标准组织培养塑料器皿获得的产率高达 10000 感染单位/细胞。监测细胞培养条件的葡萄糖消耗、乳酸产生和氨积累。葡萄糖消耗和乳酸积累与细胞生长参数密切相关。氨的产生似乎并不显著。基于病毒产率、操作简便性和线性可扩展性，使用搅拌罐生物反应器中的 293 细胞（适应于悬浮/无血清培养基或含血清培养基中的微载体）似乎可以大规模生产腺病毒。
