Instructions

Culturing human ES and iPS cells

This protocol provides a fully defined, animal-origin-free method to transition, maintain, passage, freeze/thaw, and expand human pluripotent stem cells (hPSCs) on Biolaminin 521 — enabling reproducible, single-cell culture without the need for ROCK inhibitor after adaptation.

What’s inside

  • How to transition hPSCs from feeders or other matrices onto Biolaminin 521, including initial adaptation guidelines
  • A detailed passaging workflow (dissociation, seeding densities, timing) and how to maintain hPSCs as homogeneous monolayers without spontaneous differentiation
  • Instructions for cryopreservation and thawing of hPSCs on Biolaminin 521 to ensure viability and reproducibility
  • Culture recommendations (coating concentrations, feeding schedule, confluence thresholds) to support long-term self-renewal and stable growth
Download