163 - Automated quantitative image analysis and precision selection of desired sub-population of iPS cells using cell XTM and ColonyzeTM platform

Abstract

Induced pluripotent stem (iPS) cells are being developed for a broad range of research and therapeutic applications. However, reprogrammed iPS clones are inevitably heterogeneous. Value judgements must be made regarding which clones to use or exclude. iPS labs currently make decisions by visual inspection and manual selection, which are prone to large variation and lack the standardization needed for clinical translation. Automated methods are needed that enable rapid, quantitative, repeatable and reproducible: i) imaging and quantitative analysis of iPS clones, ii) objective determination of critical quality attributes (CQAs) (e.g. morphology or markers) that predict long term biological performance and iii) precise automated selection of desired cells or removal non-desired cells. This paper describes the development and validation of Cell XTM (Figure 1), a robust robotic system that enables each of these critical aspects of iPS cell processing and analysis. Large field of view images are acquired from standard tissue culture plates (6 to 96 wells) with 1 µ/pixel resolution in 1–8 minutes, using phase-contrast or fluorescence. Integration with ColonyzeTM software enables Automated Multimodal Large field of view Image Analysis (AM-LIA) based on metrics of proliferation, migration, surface markers, or cell and colony morphology, using principles outlined in ASTM Standard F2944-12. This integration enables automated image capture, analysis and “picking” or “removal” of desired or undesired colonies based on defined and reproducible criteria (Figure 2). Cell XTM and ColonyzeTM together eliminated labor-intensive manual processing steps, and provide unprecedented rigor in the analysis and management of heterogeneous iPS cell populations. Figure 2100% picking efficiency is observed (A,B; C,D). Post picking, cells were found to maintain their viabilility and expand during the 4 day follow-up. View Large Image Figure Viewer Download Hi-res image