Characterization and transformation potential of "Synthetic" astrocytes differentiated from murine embryonic stem...

Our objective was to determine if murine embryonic stem (ES) cells, which are readily available from repositories, could be developed as a model of gliomagenesis, recognizing the difficulty in obtaining and transforming somatic astrocytes. Using a stringently controlled sequential differentiation procedure on wild type (wt) and p53+/- ES cells, we established GFAP+A2B5-synthetic astrocytes with high efficiency (>90%). The synthetic astrocytes stably express several differentiated astrocyte associated structural proteins and biochemical markers, but lacked expression of differentiated neurons and oligodendrocytes. However, in contrast to somatic differentiated astrocytes, the synthetic astrocytes expressed stem cell markers, with a transcriptome profile similar to astrocytes differentiated from neural stem cells (NSC) and somatic astrocyte cultures established from E13.5-Cortex and P4-hippocampus. In addition, the synthetic astrocytes demonstrated plasticity, with ability to dedifferentiate into neuronal and oligodendrocyte lineages. Intracranial injection of postnatal differentiated somatic astrocytes or synthetic astrocytes of either wt or p53+/- background did not grow tumors, unlike corresponding ES cells that develop teratomas. In contrast, retroviral transduction of either wt or p53+/- synthetic astrocytes and not the postnatal somatic astrocytes, with relevant oncogenes found in human malignant astrocytomas (MDM2, myr-AKT, V12H-RAS), led to intracranial high-grade undifferentiated gliomas. This study demonstrates utilization of readily available ES cells of varying genetic backgrounds to model and further our understanding of gliomagenesis. Large numbers of replenishable derivative synthetic glial lineage cells retain genetic and phenotypic characteristics of progenitor cells and thereby are more amenable to transformation by genetic aberrations involved in gliomagenesis.