This disclosure provides an improved
system for culturing human pluripotent stem cells. Traditionally, pluripotent stem cells are cultured on a layer of
mouse embryonic fibroblast feeder cells to prevent them from differentiating. In the
system described here, the role of feeder cells is replaced by defined components added to the
culture environment that support rapid proliferation without differentiation. The medium contains an isotonic buffer, a blend of essential nutrients such as
protein and lipids, and an effective
growth factor or combination of factors that promote proliferation while inhibiting differentiation. Culturing human embryonic stem cells in fresh medium on an
extracellular matrix according to this invention causes the cells to expand surprisingly rapidly, while retaining the ability to differentiate into cells representing all three embryonic germ
layers. This new culture
system allows for bulk proliferation of pPS cells for commercial production of important products for use in
drug screening and human therapy.