Ohlson S, Branscomb J, Nilsson K
Department of Natural Sciences, University of Kalmar, Sweden.
Cytotechnology. 1994;14(1):67-80. doi: 10.1007/BF00772197.
Chinese hamster ovary cells (CHO-K1) were cultivated in macroporous gelatin microcarriers (CultiSpher G and CultiSpher S) in spinner flasks and a 51 bioreactor. Near-to-confluent cultures were harvested by bead-to-bead transfer where intact microcarriers with cells were transferred from a spinner flask to another spinner flask or to the bioreactor with naked microcarrier beads. Successful bead-to-bead transfer was achieved in various split ratios. The duration of attachment seemed to be important where the direct contact of beads to each other can be achieved by intermittent stirring. Repeated transfers were performed and at least four transfers in spinner flasks were achieved. Two variations of bead-to-bead transfer were performed in the 51 bioreactor either by seeding the bioreactor with near-to-confluent beads cultivated in spinner flasks or in situ transfer by adding fresh beads to the bioreactor. As in the spinner case, attachment was achieved by intermittent stirring where donor beads were in close proximity to the acceptor beads. Again successful transfers were obtained as evidenced by the good growth on acceptor beads where cell yields were in the range of 3100-4500 cells/bead. The results suggest that bead-to-bead transfer of CHO-K1 cells can be easily performed and do provide an alternative route to applications where dissolution techniques may not offer an efficient solution.
中国仓鼠卵巢细胞(CHO-K1)在转瓶和5L生物反应器中的大孔明胶微载体(CultiSpher G和CultiSpher S)中培养。接近汇合的培养物通过珠对珠转移收获,其中带有细胞的完整微载体从一个转瓶转移到另一个转瓶或转移到装有裸微载体珠的生物反应器中。在各种分裂比例下都成功实现了珠对珠转移。附着的持续时间似乎很重要,通过间歇搅拌可以使珠子彼此直接接触。进行了重复转移,并且在转瓶中至少实现了四次转移。在5L生物反应器中进行了两种珠对珠转移变体,一种是用在转瓶中培养的接近汇合的珠子接种生物反应器,另一种是通过向生物反应器中添加新鲜珠子进行原位转移。与转瓶情况一样,通过间歇搅拌实现附着,使供体珠子靠近受体珠子。再次获得了成功的转移,受体珠子上良好的生长证明了这一点,细胞产量在3100 - 4500个细胞/珠的范围内。结果表明,CHO-K1细胞的珠对珠转移可以很容易地进行,并且确实为溶解技术可能无法提供有效解决方案的应用提供了一条替代途径。