Abstract
Purpose :
Retinal pigment epithelial (RPE) cells differentiated from human induced pluripotent stem (iPS) cells were transplanted into patients with age-related macular degeneration (Mandai et al., NEJM, 2017). Demands of cell culture methods with minimizing quality dispersion have increased in basic research and in the clinical setting. In most cases, to provide high quality cells constantly depends on the skill of the technicians having expertise in cell culture technician; however, the pressing issue is that it is difficult to teach, considering the breadth of tacit knowledge required. An automated culture by line-type robot has been indicated as a method to resolve discrepancies in quality and has helped achieve some progress in the "development" stage in the research and development (R&D) sector. On the other hand, it is difficult to introduce a line-type robot into the "research" stage of R&D owing to frequent modification of protocols. This study suggests the use of LabDroid "Maholo," a versatile humanoid robot, as a solution of these issues through the implementation of the iPS-RPE differentiation protocol in the robot.
Methods :
The differentiation protocol of the iPS-RPE cells was taught to LabDroid "Maholo." We attempted to establish a workflow to mechanically identify various parameter sets in protocols, which is the combination of time, strength, reagent concentration, etc. iPS-RPE cells thus harvested were quantified via pigmentation and biological assays such as those to quantify vascular endothelial growth factor and pigment epithelium-derived factor levels.
Results :
We installed motions of the iPS-RPE protocol into the LabDroid and achieved stable operation. Values obtained from culturing iPS-RPE cells in several parameter sets by the LabDroid and those obtained from cells cultured by cell culture experts were almost equivalent. These parameter sets were mechanically determined from our workflow.
Conclusions :
We achieved the implementation of expert skill as that of cell culture technicians in the LabDroid. Dissemination of information regarding these culture conditions and the protocol can facilitate researchers who are not adept with culturing of iPS-RPE cells can also carry out studies on iPS-RPE cells. We intend on disseminating this information to accelerate basic research on iPS and/or RPE cells.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.