A scalable and cost-effective synthetic polymer substrate that supports robust expansion and subsequent multilineage differentiation of human pluripotent stem cells (hPSCs) with defined commercial media is presented. This substrate can be applied to common cultureware and used off-the-shelf after long-term storage. Expansion and differentiation of hPSCs are performed entirely on the polymeric surface, enabling the clinical potential of hPSC-derived cells to be realized. Data presented herein includes time-of-flight secondary-ion mass spectrometry (ToF-SIMS), X-ray photoelectron spectroscopy (XPS), water contact angle (WCA) measurements, fluorescence microscopy, cell electrophysiology, nuclear magnetic resonance (NMR) spectroscopy, gel permeation chromatography (GPC). Please note that this data set is in two parts, the second part of which can be found here: http://doi.org/10.17639/nott.16