Moreira J L, Feliciano A S, Santana P C, Cruz P E, Aunins J G, Carrondo M J
Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Quimíca e Biológica (IBET/ITQB), Oeiras, Portugal.
Cytotechnology. 1994;15(1-3):337-49. doi: 10.1007/BF00762409.
Natural aggregates of Baby Hamster Kidney cells were grown in stirred vessels operated as repeated-batch cultures during more than 600 hours. Different protocols were applied to passaging different fractions of the initial culture: single cells, large size distributed aggregates and large aggregates. When single cells or aggregates with the same size distribution found in culture are used as inoculum, it is possible to maintain semi-continuous cultures during more than 600 hours while keeping cell growth and viability. These results suggest that aggregate culture in large scale might be feasible, since a small scale culture can easily be used as inoculum for larger vessels without noticeable modification of the aggregate characteristics. However, when only the large aggregates are used as inoculum, it was shown that much lower cell concentrations are obtained, cell viability in aggregates dropping to less than 60%. Under this 'selection' procedure, aggregates maintain a constant size, larger than under batch experiments, up to approximately 400 hours; after this time, aggregate size increases to almost twice the size expected from batch cultures.
将幼年仓鼠肾细胞的天然聚集体在作为重复分批培养操作的搅拌容器中培养了600多个小时。应用了不同的方案来传代初始培养物的不同部分:单细胞、大小分布不均的聚集体和大聚集体。当使用培养物中发现的具有相同大小分布的单细胞或聚集体作为接种物时,有可能在600多个小时内维持半连续培养,同时保持细胞生长和活力。这些结果表明大规模聚集体培养可能是可行的,因为小规模培养物可以很容易地用作较大容器的接种物,而聚集体特性不会有明显改变。然而,当仅使用大聚集体作为接种物时,结果表明获得的细胞浓度要低得多,聚集体中的细胞活力降至60%以下。在这种“选择”程序下,聚集体保持恒定大小,比分批实验中的大,直至约400小时;在此之后,聚集体大小增加到几乎是分批培养预期大小的两倍。