HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 2005-0392v1 24/4/865    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints/Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Luo, Y. Articles by Rao, M. S. Search for Related Content PubMed PubMed Citation Articles by Luo, Y. Articles by Rao, M. S.

CHARACTERIZATION SERIES

A Focused Microarray to Assess Dopaminergic and Glial Cell Differentiation from Fetal Tissue or Embryonic Stem Cells

^a Gerontology Research Center; Stem Cell Biology Unit, Laboratory of Neurosciences, National Institute on Aging, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland, USA;
^b Laboratory of Molecular Neurobiology, Medical Biochemistry and Biophysics, Retzius Laboratory, Karolinska Institutet, Stockholm, Sweden;
^c Buck Institute for Age Research, Novato, California, USA;
^d SuperArray Bioscience Corporation, Frederick, Maryland, USA;
^e Department of Neuroscience, Johns Hopkins University, Baltimore, Maryland, USA;
^f Invitrogen Corporation, Carlsbad, California, USA

Key Words. Human embryonic stem cells • Differentiation • Oligodendrocytes • Astrocytes • Dopamienrgic neurons

Correspondence: Mahendra S. Rao, M.D., Ph.D., Invitrogen Corporation, 1600 Faraday Road, Carlsbad, California 92008, USA. Telephone: 240-344-1781; Fax: 410-558-8249; e-mail: mahendra.rao{at}invitrogen.com

Received August 15, 2005; accepted for publication December 5, 2005.
We designed oligonucleotide gene-specific probes to develop a focused array that can be used to discriminate between neural phenotypes, identify biomarkers, and provide an overview of the process of dopaminergic neuron and glial differentiation. We have arrayed approximately 100 genes expressed in dopaminergic neurons, oligodendrocytes, and astrocytes, an additional 200 known cytokines, chemokines, and their respective receptors, as well as markers for pluripotent and progenitor cells. The gene-specific 60-mer 3' biased oligonucleotides for these 281 genes were arrayed in a 25 x 12 format based on function. Using human adult brain substantia nigra, human embryonic stem cells (ESCs), and the differentiated progeny of pluripotent cells, we showed that this array was capable of distinguishing dopaminergic neurons, glial cells, and pluripotent cells by their gene expression profiles in a concentration-dependent manner. Using linear correlation coefficients of input RNA with output intensity, we identified a list of genes that can serve as reporting genes for detecting dopaminergic neurons, glial cells, and contaminating ESCs and progenitors. Finally, we monitored NTera2 differentiation toward dopaminergic neurons and have shown the ability of this array to distinguish stages of differentiation and provide important clues to factors regulating differentiation, the degree of contaminating populations, and stage of cell maturity. We suggest that this focused array will serve as a useful complement to other large-scale arrays in routine assessment of cell properties prior to their therapeutic use.

This article has been cited by other articles:

				Y. M.-S. Tay, W.-L. Tam, Y.-S. Ang, P. M. Gaughwin, H. Yang, W. Wang, R. Liu, J. George, H.-H. Ng, R. J. Perera, et al. MicroRNA-134 Modulates the Differentiation of Mouse Embryonic Stem Cells, Where It Causes Post-Transcriptional Attenuation of Nanog and LRH1 Stem Cells, January 1, 2008; 26(1): 17 - 29. [Abstract] [Full Text] [PDF]

				M. S. Rao One Successful Series Begets Another Stem Cells, October 1, 2006; 24(10): 2160 - 2161. [Full Text] [PDF]