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Description

Millipore's Human iPS Selection Kit is a quick, easy and non-invasive method to monitor the pluripotent state of live human ESC and fully reprogrammed human iPS cells using immunocytochemistry (ICC). The Human iPS Selection Kit allows live cell imaging and identification of fully reprogrammed human iPS cells from a heterogeneous population of reprogramming intermediates and enables the live selection of human iPS cells that can be further passaged and expanded for downstream applications. The selected live stained human iPS clones maintain normal human ESC morphology and can proliferate and be passaged normally. The kit consists of the following: (1) two conjugated antibodies that are known markers of human pluripotency, Tra-I-60 and SSEA-4, (2) a negative selection marker (CD13-PE) that is specific to fibroblasts and (3) a cell permeable Hoechst nuclear dye that possesses the dual function of labeling all cells and also distinguishing partially reprogrammed cells (Hoechst Bright) from fully reprogrammed cells (Hoechst Dim) (Chan et al,)., Recent findings indicate that fully reprogrammed human iPS cells possess the following characteristics: (1) they downregulate expression of fibroblast marker, CD13 (2) they upregulate expression of pluripotent markers, SSEA-4 and TRA-1-60, (3) they silence the viral transgenes while (4) reactivating endogenous expression of Nanog and (5) they assume a HoeschstDim phenotype. These characteristics have enabled reliable identification of fully reprogrammed human iPS cells from a mixed population of partially reprogrammed cells.