Small Molecule-Assisted, Line-Independent Maintenance of Human Pluripotent Stem Cells in Defined Conditions

• DOI: 10.1371/journal.pone.0041958
• Publication Date: 2012-07-30T21:13:56Z
• Authors: Stefan Frank; Miao Zhang; Hans R. Schöler; Boris Greber
• Keywords: Chemically defined medium

Human pluripotent stem cells (hPSCs) are conventionally grown in a mouse feeder cell-dependent manner. Chemically defined culture conditions are, however, desirable not only for potential medically oriented applications but also investigating mechanisms of self-renewal and differentiation. In light the rather high complexity cost existing hPSC systems, we have systematically evaluated over 20 media ingredients. Only components that reproducibly gave beneficial effects were ultimately combined to yield simple cost-effective formulation termed FTDA. This xeno-free medium is based on mimicking factor activities present embryonic fibroblast-conditioned medium, at minimal dosages. Additionally, small molecule inhibitors BMP WNT signaling served specifically suppress typical types spontaneous differentiation seen cultures. FTDA was suitable generation human induced enabled robust long-term maintenance diverse lines including hard-to-grow ones. Comparisons with suggested reduced rates Our results imply using supportive factors concentrations may still promote preserve pluripotency hPSCs.