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

• Published in: Stem cells (Dayton, Ohio) Vol. 26; no. 6; pp. 1444 - 1453
• Main Authors: Wianny, Florence, Bernat, Agnieszka, Huissoud, Cyril, Marcy, Guillaume, Markossian, Suzy, Cortay, Véronique, Giroud, Pascale, Leviel, Vincent, Kennedy, Henry, Savatier, Pierre, Dehay, Colette
• Format: Journal Article
• Language: English
• Published: Bristol John Wiley & Sons, Ltd 01.06.2008; Alphamed Press; AlphaMed Press
• Subjects: Alkaline Phosphatase; Alkaline Phosphatase - metabolism; Animals; Blastocyst; Blastocyst - cytology; Cell Differentiation; Cell Line; Cellular Biology; Embryonic Stem Cells; Embryonic Stem Cells - cytology; Embryonic Stem Cells - enzymology; Embryonic Stem Cells - physiology; Embryonic Stem Cells - virology; Genes, Reporter; Green fluorescent protein; Green Fluorescent Proteins; Green Fluorescent Proteins - genetics; Lentivirus; Life Sciences; Macaca mulatta; Primates; Reverse Transcriptase Polymerase Chain Reaction; Rhesus monkey; Stem Cell Transplantation; Stem Cell Transplantation - methods; tau Proteins; tau Proteins - genetics; Teratoma; Teratoma - genetics; Teratoma - pathology; Transfection; Zona Pellucida; Zona Pellucida - physiology; Green Fluorescent Proteins; Cell Line; Teratoma; Genes; Blastocyst; Reporter; Macaca mulatta; Alkaline Phosphatase; Reverse Transcriptase Polymerase Chain Reaction; Lentivirus; tau Proteins; Stem Cell Transplantation; Animals; Zona Pellucida; Transfection; Cell Differentiation; Embryonic Stem Cells
• ISSN: 1066-5099; 1066-6099; 1549-4918
• DOI: 10.1634/stemcells.2007-0953

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.