(1291-D) Fully automated iPSC differentiation, expansion and maintenance.

Abstract: Induced pluripotent stem cells (iPSCs) are becoming increasingly used for drug discovery and cell therapy, yet their complex handling requirements demand time, resources and expertise. Over the years, scientists have turned to automation to alleviate the need for skilled stem cell scientists. Large robotic systems and traditional liquid handling systems have been deployed but not without limitations: large infrastructural requirements, complex setup, lack of reliability and contamination-prone open systems.

At Mytos, we’ve designed a system from the ground up that can reliably automate all common processes found in mammalian cell culture. The system interfaces fluidically with T-flasks in a closed system, avoiding exposing cells to external contaminants. Scientists can automate the culture of iPSCs from end to end such as coating, seeding, feeding, imaging, harvesting and passaging. We’ve closely mimicked manual cell culture steps by design to allow scientists without prior automation training to configure and use the system.

Beyond fluid handling, the system integrates a phase microscopy unit that allows continuous monitoring of cell growth. Combined with our cloud-based control system, scientists can check their cells directly from a web-based app. Continuous morphological information and interacting with the culture remotely at any time not only ensures robust high-quality cells by passaging at the right time but allows for retroactive analysis of the cells.

Our study aimed to demonstrate that our fully automated system was able to produce equivalent quality iPSCs to manually cultured ones. We’ve performed rigorous testing across diverse iPSC lines and culture conditions. Comprehensive assessments of cell morphology, genomic integrity, pluripotency and stemness markers were carried out.

Based on our results, we consistently demonstrated comparability with cells cultured by experienced stem cell scientists. iPSC colonies were morphologically equivalent and no differences in TRA-1-81, Nanog and SSEA4 markers were observed between manually grown iPSCs and the ones grown on the Mytos system. Samples passed Thermofisher Scientific’s Karyostat+ and PluriTestTM.

In 2023, Mytos demonstrated successful neural and cardiac differentiations using the system. Differentiated neurons were equivalent to their manual counterparts with similar morphology and expression of neuron-specific markers. As with cardiac differentiation, beating cardiomyocytes with high expression of cardiac Troponin T (cTNT) were obtained.

In summary, the system represents a significant leap forward in iPSC culture automation. Beyond time-saving, the Mytos system offers improved reproducibility, reduced contamination rates and minimal cell disturbance during handling. The device addresses shortcomings of previous systems while maintaining standards akin to manual cell culture. Mytos is glad to be lowering the barrier to iPSC research and broadening accessibility to iPSCs.