Effect of Staurosporine on Neural Differentiation of CD133+ Umbilical Cord Blood Cells

Objective: CD133+ umbilical cord blood cells were identified as a hematopoietic stem cell which has the capacity for extensive self-renewal and differentiation. The aim of this study was to identify the effect of staurosporine (STS), a wellknown protein kinase inhibitor on differentiation of CD133+ cells into neural cells. Materials and Methods: CD133+ cells were enriched by immunomagnetic beads from human mononuclear cells of umbilical cord blood and the purity of higher than 94% was achieved by flowcytometry. Induction of differentiation was performed by addition of STS (12.5, 25, and 50 n?). The differentiated cells were evaluated by immunofluorescence and RT-PCR for neuron-specific proteins and transcripts. Results: STS-treated CD133+ cells expressed mRNA transcripts for neuronspecific neurofilament protein (NFM), and several basic helix-loop-helix (bHLH) transcription factors important for early neurogenesis, including Otx2, Wnt1, and Hash1. The structural proteins characteristics of neurons including B-tubulinIII and Microtubule-Associated Protein-2 (MAP-2), were shown by immunocytochemistry. STS-treated CD133+ cells also expressed the astrocytespecific marker, glial fibrillary acidic protein (GFAP) by immunofluorescence. Conclusion: The human cord blood-derived CD133+ hematopoietic stem cells could differentiate into neural cell types of neuron-like cells and astrocytes by STS treatment.