Abstract

Generation and Characterization of Patient-Specific iPSC Lines and Gene Editing of hESCs and iPSCs

Emily Titus, Director, Technology Development , Centre for Commercialization of Regenerative Medicine (CCRM)

CCRM is a translational centre based in Toronto, Canada, focused on the development and commercialization of regenerative medicine and cell therapy technologies. An early achievement was the establishment of an iPSC production facility focused on generating high quality pluripotent cell lines from patient samples for academic researchers and clinicians. Fully operational for four years, CCRM has delivered over 100 patient-derived iPSC lines that are being used for disease modelling, and in drug screening initiatives, at Institutes across Canada. Specializing in non-integrative reprogramming technologies, CCRM has developed SOPs to reprogram many common cell types in feeder-free conditions, including dermal fibroblasts, bone marrow stromal cells, cord and peripheral blood, and endothelial cells. In addition to cell health and quality assays, which include karyotyping, absence of mycoplasma, STR and post-thaw viability, iPSC lines are characterized by flow cytometry and qPCR, and tested using directed differentiation protocols to screen for pluripotent potential. Most recently, CCRM has established gene editing in hESCs and iPSCs using both TALENs and CRISPR/Cas9 to generate reporter cell lines and perform gene correction on patient-derived iPSCs. This presentation will focus on the technical challenges and associated solutions implemented at CCRM to establish these workflows.