Derivation and cloning of a novel rhesus embryonic stem cell line stably expressing tau-green fluorescent protein.
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.
Wianny
Florence
F
Institut National de la Santé et de la Recherche Médicale, U846 Stem Cell and Brain Research Institute, Bron, France.
Bernat
Agnieszka
A
Huissoud
Cyril
C
Marcy
Guillaume
G
Markossian
Suzy
S
Cortay
Véronique
V
Giroud
Pascale
P
Leviel
Vincent
V
Kennedy
Henry
H
Savatier
Pierre
P
Dehay
Colette
C
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology / Q000201:enzymology / Q000502:physiology* / Q000821:virology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics*; D016086:Lentivirus; D008253:Macaca mulatta; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033581:Stem Cell Transplantation / Q000379:methods; D013724:Teratoma / Q000235:genetics / Q000473:pathology; D014162:Transfection; D015044:Zona Pellucida / Q000502:physiology; D016875:tau Proteins / Q000235:genetics*
D016875:tau Proteins; D049452:Green Fluorescent Proteins; D000469:Alkaline Phosphatase