Study on pipetting motion optimization of automatic spheroid culture system for spheroid formation

This research group has established a technology for producing a three-dimensional cell constructed using only the cell itself. This technology uses a property in which the spheroids fuse with each other. We developed a system that automates the spheroid production process to obtain reproducible spheroids and suppress variation factors that occur from human operation. However, it has become clear that the dispersion oc-curs in the diameter depending on the number of cells of the spheroid even if the cells are handled in the same manner. The purpose of this research is to examine an appropriate pipetting motion in accordance with the number of cells of the spheroid to be produced. Rabbit mesenchymal stem cells (rMSCs) are used as the ob-jects. The number of cells was set to 2 × 10⁴, 3 × 10⁴, and 4 × 10⁴ cells/well, and the passage number as 7. The appearance of spheroids cultured using the motion programmed in accordance with each number of cells was observed every 24 hours for 5 days after seed-ing. The results of the analysis indicate that the opti-mum motion in each number of cells has been successfully specified, and reproducible spheroids have been successfully produced.