Derivation and Cloning of a Novel Rhesus ES cell line stably expressing Tau-GFP

¹ Inserm, U846, Stem Cell and Brain Research Institute, 18 Avenue Doyen Lepine, 69500 Bron, France; Université de Lyon, Lyon 1, UMR-S 846, 69003 Lyon, France
² Inserm, U846, Stem Cell and Brain Research Institute, 18 Avenue Doyen Lepine, 69500 Bron, France; Université de Lyon, Lyon 1, UMR-S 846, 69003 Lyon, France; Hospices civils de Lyon, Department of Obstetrics and Gynecology, Hôpital de la Croix Rousse, 93 Grande rue de la Croix- Rousse, 69004 LYON, France
³ Inserm, U846, Stem Cell and Brain Research Institute, 18 Avenue Doyen Lepine, 69500 Bron, France; Université de Lyon, Lyon 1, UMR-S 846, 69003 Lyon, France; PrimaStem, Inra USC 2008, 18 Avenue Doyen Lepine, 69500 Bron, France

^* To whom correspondence should be addressed. E-mail: Colette.Dehay{at}inserm.fr.

	Abstract

Embryonic stem cells (ESC) have the ability of indefinite self-renewal and multilineage differentiation, and 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 to visualize cell bodies, dendrites and axons in exquisite details. Here, we report the first rhesus monkey ESC line that ubiquituously 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 ES cells. 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 ES cell 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 precisely be mapped in vivo after grafting in the adult rat brain.