We have developed for hESc a new 3-Stage directed differentiation protocol (DDP) to generate chondrocytes, the specialized cells that form cartilage tissue. The protocol is segmented into stages that mimic the developmental processes that occur in cell lineage specification during embryogenesis. Pluripotent hESc, are established as feeder-free cultures4 prior to DDP being carried out in monolayer culture on a sequence of matrix protein-coated surfaces with a chemicallydefined medium sequentially supplemented with growth factors to directdifferentiation through developmental intermediate cell populations of primitive streak/mesendoderm (Stage 1), mesoderm (Stage 2) and then chondrocytes (Stage 3) (Figure 1). This unique protocol is highly efficient, scalable and completely chemically-defined, thus making it appropriate for translation towards clinical applications of chondrocytes, as well as providing a defined system for characterizing the molecular mechanisms regulating hESc differentiation.
Copyright: © 2012 Rachel A. Oldershaw, Melissa A. Baxter, Emma T. Lowe, Nicola Bates, Lisa M. Grady, Francesca Soncin, Daniel R. Brison, Timothy E. Hardingham, and Susan J. Kimber.