Research Paper Volume 14, Issue 10 pp 4445—4458
A real-time pluripotency reporter for the long-term and real-time monitoring of pluripotency changes in induced pluripotent stem cells
Figure 1. Generation of mouse Oct4-EGFP iPSCs from mouse embryonic fibroblasts (MEFs). (A) A schematic diagram of the reprogramming protocol used. (B) Typical Oct4-GFP+ miPSC colonies were initially observed around day 14. (C) The Oct4-GFP+ miPSCs sustain 30 generations and homogenous self-renewal under conventional mESC growth condition. (D) The long-term expanded iPSCs grow as compact and domed colonies that express strong alkaline phosphatase (ALP). (E) These miPSCs express typical pluripotency markers, and GFP (green) was shown to be colocalized with SSEA1(red), Sox2 (red), Oct4 (red) and Nanog(red). (F) RT-PCR analysis of endogenous pluripotency gene expression in Oct4-GFP+ miPSCs. Scale bar: (B, C) 100μm and (D, E) 50μm.