Horiguchi Ikki, Nagate Hotaka, Sakai Yasuyuki
Division of Chemical Engineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama-cho, Toyonaka, Osaka 560-8531, Japan.
Department of Chemical System Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
J Biosci Bioeng. 2023 May;135(5):411-416. doi: 10.1016/j.jbiosc.2023.02.007. Epub 2023 Mar 14.
Suspension cultures are widely used for cell expansion in regenerative medicine and production. Shaking culture is one of the useful suspension culture methods that ensures gentle agitation. There are other shaking methods, including orbital shaking, reciprocating, and rocking; however, optimizing the shaking conditions for each method to meet cell culture requirements is time-consuming. In this study, we used a particle-tracking-based strategy for optimizing the agitation conditions. When the average accelerations of aggregates were calculated, high acceleration occurred periodically, and acceleration of the aggregates in orbital shaking was stable. Furthermore, the number of dead cells correlated with the average time of acceleration. We observed that cell growth was ideally maintained by factors such as optimal acceleration, aggregate formation, and cell death. These results indicate that the image-based analyses of aggregates help optimize the agitation conditions for the shaking suspension culture of induced pluripotent stem cells (iPSCs).
悬浮培养在再生医学和生产中被广泛用于细胞扩增。振荡培养是确保轻柔搅拌的有用悬浮培养方法之一。还有其他振荡方法，包括轨道振荡、往复振荡和摇摆振荡；然而，针对每种方法优化振荡条件以满足细胞培养要求是耗时的。在本研究中，我们使用基于粒子追踪的策略来优化搅拌条件。当计算聚集体的平均加速度时，会周期性地出现高加速度，并且轨道振荡中聚集体的加速度是稳定的。此外，死细胞数量与平均加速时间相关。我们观察到细胞生长通过最佳加速度、聚集体形成和细胞死亡等因素得到理想维持。这些结果表明，基于图像的聚集体分析有助于优化诱导多能干细胞（iPSC）振荡悬浮培养的搅拌条件。
