Effect of staurosporine on neural differentiation of CD133[+] umbilical cord blood cells

ABSTRACT

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 well-known protein kinase inhibitor on differentiation of CD133[+] cells into neural cells. 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 nM]. The differentiated cells were evaluated by immunofluorescence and RT-PCR for neuron-specific proteins and transcripts. STS-treated CD133[+] cells expressed mRNA transcripts for neuron-specific 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 beta-tubulinlll and Microtubule-Associated Protein-2 [MAP-2], were shown by immunocytochemistry. STS-treated CD133[+] cells also expressed the astrocyte-specific marker, glial fibrillary acidic protein [GFAP] by immunofluorescence. The human cord blood-derived CD133[+] hematopoietic stem cells could differentiate into neural cell types of neuron-like cells and astrocytes by STS treatment