(T1116) UTILIZING AUTOMATION PLATFORTMS TO ENABLE SCALABLE AND REPRODUCIBLE IPSC GENOME ENGINEERING AND DIFERENTIATION FOR DOWNSTREAM ASSAY DEVELOPMENT AND DISEASE MODELING
Senior Scientist Regeneron Tarrytown, New York, United States
Abstract: iPSCs provide a unique platform for studying disease through our ability to both genetically engineer and differentiate them into relevant cell types. This singular platform brings with it special challenges in both the time and labor of performing the genetic engineering as well as consistency in the differentiation of iPSCs into the desired cell types. Consistency and reproducibility independent of the user has always been an elusive goal that can be mitigated by moving to automated platforms. One of the major challenges in the automation of engineered iPSCs is generating clean clonal populations that can be expanded, screened, and banked for future applications including differentiation. Using various inhibitors, medias, and extra cellular matrices, we have optimized a protocol for single cell ‘printing’ or sorting of cells post-electroporation into multi well plates with a high rate of clone survival. These clones are then expanded, banked, and screened for clonality before being used for downstream differentiation all within an automated platform. This not only alleviates ergonomic strain on the scientists involved but we also have shown this method reduces overall time and handling of the iPSCs leading to a faster pipeline. The second arm of our automation platforms is differentiation of iPSCs to various cells of interest. As differentiations can be challenging and labor intensive, typically requiring a working group per lineage, automating the differentiation provided us the opportunity to increase the number of differentiations being performed. We have previously adapted several protocols from the literature to go down the mesendodermal pathway to support various groups and allow for downstream assay development and functional characterization. Combining our automation and differentiation protocols with our already existing ability to genetically engineer genetic variants via CRISPR/Cas9, these platforms could be used to increase the number of generated clones as well as the scale and number of differentiations being performed.
