This application note demonstrates that human pluripotent stem cells (hPSCs) can be efficiently transitioned from high-protein, albumin-rich media such as mTeSR™ Plus to the fully defined, animal-free HiDef® B8 medium using either direct or gradual adaptation workflows. The study provides practical, validated procedures for both adherent ESCs and iPSCs, including a five-day stepwise approach that maintains healthy morphology, stable growth, and robust pluripotency throughout the transition. Cells adapted to HiDef-B8 exhibit the expected shift to a looser, “continent-like” colony morphology while retaining high viability and expression of pluripotency markers, with genomic stability supported over extended passaging as previously published.

The guide further highlights how Ready-CEPT™ enhances survival and consistency during enzymatic or single-cell workflows, improving outcomes during adaptation and enabling gene-editing and clonal expansion applications. Key considerations, including recommended hypoxic culture conditions, ideal confluence, passaging strategies, and substrate compatibility, ensure reproducible performance across diverse hPSC lines. Developed to overcome the variability, cost, and animal-derived components found in legacy media systems, HiDef-B8 offers a chemically defined, serum-free platform optimized for long-term, weekend-free adherent hPSC maintenance. By simplifying workflows, reducing undefined proteins, and enabling seamless movement toward translational and cGMP-aligned processes, HiDef-B8 provides researchers with a reliable, scalable foundation for modern human pluripotent stem cell culture.