Cells Using Recombinant E8 Fragments of Human Laminin Isoforms

摘要

This unit describes a protocol for efficient expansion of human pluripotent stem cells (hPSCs). A key feature of this method is subculture of hPSCs by singlecell dissociation passaging on substrates coated with recombinant E8 fragments of human laminin isoforms (LM-E8s). LM-E8s, provide superior adhesion over intact laminin isoforms and Matrigel. Single hPSCs seeded on LM-E8s show accelerated migration and rapid reconstruction of clusters, resulting in robust survival and proliferation. This protocol yields 200-fold more hPSCs than conventional subculture methods in 1 month of culture. Furthermore, this protocol can be easily adapted to most hPSC lines in combination with the use of various xeno-free, defined culture media, and large-scale expansion of hPSCs is easily achievable to facilitate the practical applications of hPSCs. This unit describes a protocol for efficient and robust expansion of human pluripotent stem cells (hPSCs), including human embryonic stem cells and human induced pluripotent stem cells, by single-cell dissociation passaging. hPSCs are usually maintained by colony dissociation passaging.