Unraveling the Signaling Web in Stem Cells for Regenerative Medicine with Simple Western

Simple Western™ is an advanced capillary immunoblotting platform that addresses the limitations of current methods for protein expression and cell signaling pathway analysis in stem cells for regenerative medicine. Here, we used the advanced automated Western blot analysis of Simple Western to monitor iPSC differentiation into three primary germ layers of human embryos. Using GMP proteins and GMP small molecules, we differentiated iPSCs into endoderm, ectoderm, and mesoderm layers. As these cells differentiated, we applied high sensitivity multiplex assays on Simple Western to reveal and quantify changes in the expression of pluripotent factors and cell signaling proteins that regulate stem cell differentiation. With only 3 μL of starting material, we show how Simple Western has unrivaled multiplex flexibility, sensitivity, and reproducibility for accurate protein expression measurements that scale with manufacturing workflows for stem cell therapy and regenerative medicine.