Scalable 96-well Plate Based iPSC Culture and Production Using a Robotic Liquid Handling System.

摘要: Continued advancement in pluripotent stem cell culture is closing the gap between bench and bedside for using these cells regenerative medicine, drug discovery safety testing. In order to produce derived biopharmaceutics tissue engineering transplantation, a cost-effective cell-manufacturing technology essential. Maintenance of pluripotency stable performance downstream applications (e.g., differentiation) over time paramount large scale production. Yet that can be difficult achieve especially if are cultured manually where operator introduce significant variability as well prohibitively expensive scale-up. To enable high-throughput, large-scale production remove influence novel protocols bench-top multi-channel liquid handling robot were developed require minimal technician involvement or experience. With human induced (iPSCs) feeder-free conditions directly from frozen stock maintained 96-well plates. Depending on line desired scale-up rate, easily determine when passage based series images showing optimal colony densities splitting. Then necessary reagents prepared perform split new plates without centrifugation step. After 20 passages (~3 months), two iPSC lines karyotypes, expressed markers, differentiated into cardiomyocytes with high efficiency. The system subsequent high-throughput screening differentiation genetic manipulation designed This will reduce labor technical burden numbers identical myriad applications.