Identification and Characterization of iPS Clones via Automated Live-Imaging and In-Process Analysis for Quality Control using High Throughput Robotic System

ConclusionWe used DF6-9-9T.B hiPSC cell line, reprogrammed skin fibroblasts and peripheral blood mononuclear cells to develop our image processing parameters and picking and weeding protocols. To define and quantify critical quality attributes (CQAs) of “completely” reprogrammed iPS cells in systematic repeatable and reproducible manner, the Cell XTMPlatform is integrated with ColonyzeTM4.0, a very user-friendly WindowsTM-based automated quantitative cell and colony analysis software. The principles and nomenclature enabling this approach are outlined in ASTM Standard F2944-12. To automate manipulation of iPS cells in rapid, precise, repeatable and rigorously documented manner, the Cell XTMdevice integrates imaging, cell-manipulation (biopsy, pick, weed), fluid handling, and motion control systems.Using the Cell XTMgraphical user-interface, imaging, media changes and automated image processing were performed as illustrated in Figure 1. Picking operations were effective, and transplanted cells continued to proliferate effectively with morphology that is unchanged from that of the original colony. Automated image processing will display output metrics (colony area, the x,y,z-centroid for each colony, colony density, roundness, perimeter etc), as well as highlight the periphery of individual colonies or cell boundaries in the montage image. Based on these metrics, one can quantitatively define the cells or colonies of interest and selectively pick and transfer the cells for ...
Source: Cytotherapy - Category: Cytology Source Type: research