| Abstract: |
Recent reports have shown efficient differentiation of both mouse and monkey ES into a variety of CNS derivatives using stromal derived inducing activity (SDIA). Using SDIA of bone marrow-derived stromal cells dopaminergic neurons originating from mouse ES were obtained in our lab and used for experimental cell replacement in Parkinsonian rat providing evidence that the SDIA-derived DA precursor function in vivo. Human embryonic stem (HES) cells provide a potentially unlimited source for therapeutic cell replacement. We have established conditions for the differentiation of HES cells into dopaminergic precursors and neurons using SDIA of various wild-type and transgenic stromal feeders in combination with sequential growth factor regimens including SHH, FGF8, AA, BDNF and low oxygen cell culture. Here we present data on the in vitro characterization of HES-derived progeny both during and after their differentiation and maturation into dopamine neurons. Genome-wide gene expression profiles were obtained for each differentiation stage. In vivo data are presented on short time survival and stability of phenotype after transplantation into the adult rat brain. Further functional characterizations in vitro and in vivo are ongoing. |
