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EMBRYONIC STEM CELLS

Derivation and Cloning of a Novel Rhesus Embryonic Stem Cell Line Stably Expressing Tau-Green Fluorescent Protein

^aInstitut National de la Santé et de la Recherche Médicale, U846 Stem Cell and Brain Research Institute, Bron, France;
^bUniversité de Lyon, UMR-5 846, Lyon, France;
^cDepartment of Obstetrics and Gynecology, Hôpital de la Croix Rousse, Hospices Civils de Lyon, Lyon, France;
^dPrimaStem, INRA USC 2008, Bron, France

Key Words. Embryonic stem cells • Rhesus monkey • Green fluorescent protein

Correspondence: Colette Dehay, Ph.D., INSERM, U846, Stem Cell and Brain Research Institute, 18 Avenue Doyen Lepine, 69500 Bron, France. Telephone: 00-33-4-72-91-34-71; Fax: 00-33-4-72-91-34-61; e-mail: Colette.Dehay{at}inserm.fr

Received November 14, 2007; accepted for publication March 7, 2008.
First published online in STEM CELLS EXPRESS   March 20, 2008.

Embryonic stem cells (ESC) have the ability of indefinite self-renewal and multilineage differentiation, and they carry great potential in cell-based therapies. The rhesus macaque is the most relevant preclinical model for assessing the benefit, safety, and efficacy of ESC-based transplantations in the treatment of neurodegenerative diseases. In the case of neural cell grafting, tracing both the neurons and their axonal projections in vivo is essential for studying the integration of the grafted cells in the host brain. Tau-Green fluorescent protein (tau-GFP) is a powerful viable lineage tracer, allowing visualization of cell bodies, dendrites, and axons in exquisite detail. Here, we report the first rhesus monkey ESC line that ubiquitously and stably expresses tau-GFP. First, we derived a new line of rhesus monkey ESC (LYON-ES1) that show marker expression and cell cycle characteristics typical of primate ESCs. LYON-ES1 cells are pluripotent, giving rise to derivatives of the three germ layers in vitro and in vivo through teratoma formation. They retain all their undifferentiated characteristics and a normal karyotype after prolonged culture. Using lentiviral infection, we then generated a monkey ESC line stably expressing tau-GFP that retains all the characteristics of the parental wild-type line and is clonogenic. We show that neural precursors derived from the tau-GFP ESC line are multipotent and that their fate can be precisely mapped in vivo after grafting in the adult rat brain.

Disclosure of potential conflicts of interest is found at the end of this article.