Adhesive Signature-based, Label-free Isolation of Human Pluripotent Stem Cells

The ability to efficiently isolate undifferentiated human induced pluripotent stem cells (UD-hiPSCs) as colonies from contaminating non-pluripotent cells is a crucial step in the stem cell field to maintain hiPSC survival, purity, and karyotype stability. Here we demonstrate significant differences in ‘adhesive signature’ among UD-hiPSCs, parental cells, partially reprogrammed cells, and differentiated progeny. The distinct adhesive signature of hiPSCs was exploited to rapidly (~10 min) and efficiently isolate fully reprogrammed bona fide hiPSCs as intact colonies from heterogeneous reprogramming cultures and differentiated progeny using microfluidics. hiPSCs were isolated in a label-free fashion and enriched to > 95–99% purity and survival without adversely affecting the transcriptional profile, differentiation potential or karyotype of the pluripotent cells. This rapid and label-free strategy is applicable to isolate UD-hPSCs (hiPSCs, hESCs) from heterogeneous cultures during reprogramming and routine cultures and can be expanded to purify stem cells of specific lineages, such as neurons and cardiomyocytes.