Research report High-level expression of functional chemokine receptor CXCR4 on human neural precursor cells Hsiao T. Ni a , Shuxian Hu b , Wen S. Sheng b , Judy M. Olson a , Maxim C.-J. Cheeran b , Anissa S.H. Chan a , James R. Lokensgard b , Phillip K. Peterson b, * a Stem Cell Group, R&D Systems, Inc., Minneapolis, MN 55413, USA b Minneapolis Medical Research Foundation and the University of Minnesota Medical School, Minneapolis, MN 55404, USA Accepted 6 June 2004 Available online 8 August 2004 Abstract Neural precursor cells (NPCs) are self-renewing, multipotent progenitors that give rise to neurons, astrocytes and oligodendrocytes in the central nervous system (CNS). Fetal NPCs have attracted attention for their potential use in studying normal CNS development. Several studies of rodent neural progenitors have suggested that chemokines and their receptors are involved in directing NPC migration during CNS development. In this study, we established a consistent system to culture human NPCs and examined the expression of chemokine receptors on these cells. NPCs were found to express the markers nestin and CD133 and to differentiate into neurons, astrocytes and oligodendrocytes at the clonal level. Flow cytometry and RNase protection assay (RPA) indicated that NPCs express high levels of CXCR4 and low levels of several other chemokine receptors. When examined using a chemotaxis assay, NPCs were able to respond to CXCL12/SDF-1a, a ligand of CXCR4. Treatment with anti-CXCR4 antibody or HIV-1 gp120 abolished the migratory response of NPCs towards CXCL12/SDF-1a. These findings suggest that CXCR4 may play a significant role in directing NPC migration during CNS development. D 2004 Elsevier B.V. All rights reserved. Theme: Development and regeneration Topic: Cell differentiation and migration Keywords: Neural precursor cell; Differentiation; Chemokine receptor 1. Introduction Chemokines and their cognate receptors were initially associated with the trafficking of leukocytes in physiolog- ical immune surveillance and with inflammatory cell recruitment in different diseases [45,48]. Subsequently, they have been found to play a critical role in hematopoietic development by regulating migration, proliferation, differ- entiation and survival of human and murine hematopoietic stem and progenitor cells [1,3,24,27,37,56]. The involve- ment of the chemokine/chemokine receptor family in central nervous system (CNS) development has recently been identified, but is still poorly understood and remains to be elucidated. Neural precursor cells (NPCs) are self-renewing, multi- potent cells that give rise to neurons, astrocytes and oligodendrocytes in the CNS [5,10,33]. In recent years, in vitro methods have been developed which permit the expansion and differentiation of multipotent fetal NPCs in culture [5,7,19,43,44,50,54]. NPCs can be grown in the presence of mitogen, as either monolayer cultures or as free floating spherical aggregates termed dneurospheres.T Fol- lowing expansion, NPCs can be induced to differentiate by withdrawal of mitogenic agent or by exposure of the cells to other factors that cause them to develop along specific lineages. While much work is required to characterize NPCs 0165-3806/$ - see front matter D 2004 Elsevier B.V. All rights reserved. doi:10.1016/j.devbrainres.2004.06.015 * Corresponding author. Department of Medicine, Medical School and Hennepin County Medical Center, University of Minnesota, 701 Park Avenue South, Minneapolis, MN 55415, United States. Tel.: +1 612 873 2877; fax: +1 612 904 4299. E-mail address: peter137@umn.edu (P.K. Peterson). Developmental Brain Research 152 (2004) 159 – 169 www.elsevier.com/locate/devbrainres