Cardiomyocytes from human pluripotent stem cells: from laboratory curiosity to industrial biomedical platform

Cardiomyocytes from human pluripotent stem cells (hPSCs-CMs) could revolutionise biomedicine. Global burden of heart failure will soon reach USD $90bn, while unexpected cardiotoxicity underlies 28% of drug withdrawals. Advances in hPSC isolation, Cas9/CRISPR genome engineering and hPSC-CM differentiation have improved patient care, progressed drugs to clinic and opened a new era in safety pharmacology. Nevertheless, predictive cardiotoxicity using hPSC-CMs contrasts from failure to almost total success. Since this likely relates to cell immaturity, efforts are underway to use biochemical and biophysical cues to improve many of the ~ 30 structural and functional properties of hPSC-CMs towards those seen in adult CMs. Other developments needed for widespread hPSC-CM utility include subtype specification, cost reduction of large scale differentiation and elimination of the phenotyping bottleneck. This review will consider these factors in the evolution of hPSC-CM technologies, as well as their integration into high content industrial platforms that assess structure, mitochondrial function, electrophysiology, calcium transients and contractility. This article is part of a Special Issue entitled: Cardiomyocyte Biology: Integration of Developmental and Environmental Cues in the Heart edited by Marcus Schaub and Hughes Abriel

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