Inui Takumi, Mei Jiyang, Imashiro Chikahiro, Kurashina Yuta, Friend James, Takemura Kenjiro
Department of Mechanical Engineering, Keio University, Yokohama 223-8522, Japan.
Lab Chip. 2021 Apr 7;21(7):1299-1306. doi: 10.1039/d0lc01293a. Epub 2021 Mar 18.
Regenerative medicine and drug development require large numbers of high-quality cells, usually delivered from in vitro culturing. During culturing, the appearance of unwanted cells and an inability to remove them without damaging or losing most if not all the surrounding cells in the culture reduce the overall quality of the cultured cells. This is a key problem in cell culturing, as is the inability to sample cells from a culture as desired to verify the quality of the culture. Here, we report a method to locally remove cells from an adherent cell culture using a 100.4 MHz focused surface acoustic wave (SAW) device. After exposing a plated C2C12 mouse myoblast cell culture to phosphate buffered solution (PBS), ultrasound from the SAW device transmitted into the cell culture via a coupling water droplet serves to detach a small grouping of cells. The cells are removed from an area 6 × 10 mm, equivalent to about 12 cells, using a SAW device-Petri dish water gap of 1.5 mm, a PBS immersion time of 300 s, and an input voltage of 75 V to the SAW device. Cells were released as desired 90% of the time, releasing the cells from the target area nine times out of ten runs. In the one trial in ten that fails, the cells partially release and remain attached due to inter-cellular binding. By making it possible to target and remove small groups of cells as desired, the quality of cell culturing may be significantly improved. The small group of cells may be considered a colony of iPS cells. This targeted cell removal method may facilitate sustainable, contamination-free, and automated refinement of cultured cells.
再生医学和药物开发需要大量高质量的细胞，通常通过体外培养来提供。在培养过程中，不需要的细胞出现，并且在不损害或不损失培养物中大部分（如果不是全部）周围细胞的情况下无法将其去除，这降低了培养细胞的整体质量。这是细胞培养中的一个关键问题，同样存在的问题是无法根据需要从培养物中取样细胞以验证培养物的质量。在此，我们报告一种使用100.4 MHz聚焦表面声波（SAW）装置从贴壁细胞培养物中局部去除细胞的方法。将接种的C2C12小鼠成肌细胞培养物暴露于磷酸盐缓冲溶液（PBS）后，来自SAW装置的超声波通过耦合水滴传输到细胞培养物中，用于分离一小群细胞。使用SAW装置与培养皿的水隙为1.5 mm、PBS浸泡时间为300 s以及SAW装置的输入电压为75 V，从6×10 mm的区域去除细胞，相当于约12个细胞。细胞在90%的时间内按预期释放，即在十次运行中有九次从目标区域释放细胞。在十次试验中有一次失败的情况下，细胞会部分释放并由于细胞间结合而保持附着。通过能够根据需要靶向并去除小群细胞，细胞培养的质量可能会显著提高。这一小群细胞可被视为诱导多能干细胞集落。这种靶向细胞去除方法可能有助于实现培养细胞的可持续、无污染和自动化提纯。
