Development of a robust workflow for cloning iPSCs should have immediate potential to impact standards, consistency, and confidence of clonality between different laboratories in both academic and pharma research.
In this webinar, we demonstrate that numerous hiPSC cell lines can be successfully subcloned in a robust and automated fashion using the VIPS™ single cell cloning instrument in combination with a new soluble matrix called MatriClone™ (commercially available from Solentim), thereby reducing time and cost while most importantly maintaining the integrity of clonal biology.
Firstly, we show that the VIPS plus MatriClone combination results in a several-fold improvement in clonal colony outgrowth of single seeded hiPSCs when compared with manual LD. Secondly, a number of hiPSC subclones were identified from presumed healthy and disease-affected backgrounds using daily whole well imaging on the VIPS and selected for expansion approximately 10-14 days post-seeding. Expression of pluripotency was confirmed for each of the sub-cloned lines. Finally, we demonstrate that subclones maintained genomic integrity after extended culture and successfully differentiated into mixed cortical neurons.
We will discuss how future translation of such a workflow could positively effect GMP standards and re-establish expectations with the regulatory bodies for the development and production of advanced cell models, cellular diagnostics, and cell therapeutics, including clonality documentation to accompany future IND submissions.
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