Highly Efficient Reprogramming and Characterization of Induced Pluripotent Stem Cells by Using a Microwell Array

ABSTRACT

Reprogramming of human somatic cells to induced pluripotent stem cells (iPSCs) enables the possibility of generating patient-specific cells. However, the low efficiency issue associated with iPSCs generation has limited iPSCs usage in research and clinical applications. In this study, we developed a high efficiency system to generate iPSCs by using a polydimethylsiloxane stencil. This device could be applied to the localization and reprogramming of human fibroblasts. Herein, a well-defined culture system based on a stencil, which supported efficient reprogramming of fibroblasts into iPSCs with 2–4 fold increase in efficacy over conventional methods, is presented. Subsequently, we prepared a multiple analysis system, which used a micro-patterned scissile microarray to characterize iPSCs. The results showed that iPSCs could be cultured into micro-patterns in a precisely controlled manner on the scissile poly(ethylene terephthalate) sheet, which was cut into pieces for subsequent analyses, indicating that this method allows multiple analyses to establish iPSC pluripotency in the same sample. Our approach provides a simple, cost-effective, but highly efficient system for the generation and characterization of iPSCs, and will serve as a powerful tool for establishing patient- and disease-specific pluripotent stem cells.