Novel platform for fully automated generation and expansion of highly standardized iPS cells

The medical prospects of human induced pluripotent stem cells (hiPSCs) have created an urgent need for standardized and automated processes for reprogramming and expansion of hiPSC lines from large patient cohorts. This demand can be met by the StemCellFactory (www.stemcellfactory.de), an automated platform that runs and controls all required cell culture steps, ranging from adult human dermal fibroblast expansion via feeder-free, Sendai virus-based reprogramming to clonal selection and enzyme-free expansion of the obtained hiPSC clones and lines. The platform requires a highly flexible control software with extensive data management, robust device communication and a metrology based process control. For this purpose, a control software was developed, which allows to run the platform completely automatically. The measurement of confluence in regular time intervals of every well in a microtiter plate is done by high speed microscopy that was developed for the StemCellFactory platform. To achieve a complete automated run of microtiter plates with patient specific cells, threshold based decision logics are implemented that compute and translate the acquired data during the process and adapts the workflow according to the measured results. The automated process was implemented on microtiter plates and scheduled. Eventually, extensive biological validation was performed to confirm that automatically expanded hiPSCs remain pluripotent upon automated long-term (10 passages) cultivation. In summary, our data show that analysis of in-process generated data largely facilitates automation of highly dynamic cell culture processes