High Efficiency Single Cell Cloning Of hiPSCs While Ensuring Maintenance Of Phenotype

The inability to robustly manipulate hiPSCs as single cells has remained a significant biological and technical hurdle. Current techniques rely on inefficient methods such as limiting dilution (LD), colony picking and fluorescence-activated cell sorting (FACS), which are time consuming, expensive and incompatible with the sensitivity of hiPSCs. Furthermore, these methods provide insufficient documented evidence of clonality.

This application note describes how several different hiPSC lines have been successfully subcloned in a robust and automated fashion using the VIPS instrument in conjunction with optimized culture conditions. This method has been shown to reduce time and cost while most importantly maintaining the cell phenotype and genomic integrity as well as providing documented evidence of cell line clonality.

This application note describes a simple and robust workflow for the single cell cloning of hiPSCs using the platform technology combination of VIPS and MatriClone. Use of MatriClone, an animal component-free matrix based on laminin, in solution for the adaptation steps, in the reservoir for the seeding and in the media post- seeding, removes the need for pre-coating plates or the use of undefined matrices.