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a School of Biological and Biomedical Science, University of Durham, Science Laboratories, Durham, United Kingdom;
b Department of Cellular Neuroscience, Wyeth-Neuroscience, Monmouth Junction, New Jersey, USA
Key Words. Embryonal carcinoma • Stem cell • Human • Neurogenesis • Gene transcription • Microarray
Stefan Alexander Przyborski, Ph.D., School of Biological and Biomedical Science, University of Durham, South Road, Durham, DH1 3LE, United Kingdom. Telephone: 44-0-191-374-3341; Fax: 44-0-191-374-2417; e-mail: stefan.przyborski{at}durham.ac.uk
Pluripotent stem cell lines can be induced to differentiate into a range of somatic cell types in response to various stimuli. Such cell-based systems provide powerful tools for the investigation of molecules that modulate cellular development. For instance, the formation of the nervous system is a highly regulated process, controlled by molecular pathways that determine the expression of specific proteins involved in cell differentiation. To begin to decipher this mechanism in humans, we used oligonucleotide microarrays to profile the complex patterns of gene expression during the differentiation of neurons from pluripotent human stem cells. Samples of mRNA were isolated from cultured NTERA2 human embryonal carcinoma stem cells and their retinoic-acid-induced derivatives and were prepared for hybridization on custom microarrays designed to detect the expression of genes primarily associated with the neural lineage. In response to retinoic acid, human NTERA2 cells coordinately regulate the expression of large numbers of neural transcripts simultaneously. Transcriptional profiles of many individual genes aligned closely with expression patterns previously recorded by developing neural cells in vitro and in vivo, demonstrating that cultured human pluripotent stem cells appear to form neurons in a conserved manner. These experiments have produced many new expression data concerning neuronal differentiation from human stem cells in vitro. Of particular interest was the regulated expression of Pax6 and Nkx6.1 mRNA and the absence of Pax7 transcription, indicating that neurons derived from NTERA2 pluripotent stem cells are characteristic of neuroectodermal cells of the ventral phenotype.
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