Isolation of a Primate Embryonic Stem Cell Line

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 92; no. 17; pp. 7844 - 7848
• Main Authors: Thomson, James A., Kalishman, Jennifer, Golos, Thaddeus G., Durning, Maureen, Harris, Charles P., Becker, Robert A., Hearn, John P.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 15.08.1995; National Acad Sciences; National Academy of Sciences
• Subjects: alpha-Fetoproteins - biosynthesis; Animals; Antigens, Surface - analysis; Base Sequence; Biology; Blastocyst - cytology; Blastocyst - physiology; Cell Differentiation; Cell Line; Cell lines; Cell Transformation, Neoplastic; Cellular differentiation; Culture Techniques - methods; DNA Primers; Embryo, Mammalian; Embryonic cells; Embryonic stem cells; Embryos; Epithelial cells; Female; Germ cells; Glyceraldehyde-3-Phosphate Dehydrogenases - biosynthesis; Humans; Intestines; Karyotyping; Macaca mulatta; Male; Mice; Mice, SCID; Molecular Sequence Data; Monkeys & apes; Neurons; Ovulation; Pluripotent stem cells; Polymerase Chain Reaction; primates; Stem cells; Stem Cells - cytology; Stem Cells - physiology; Transplantation, Homologous; X Chromosome; Y Chromosome
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.92.17.7844

Embryonic stem cells have the ability to remain undifferentiated and proliferate indefinitely in vitro while maintaining the potential to differentiate into derivatives of all three embryonic germ layers. Here we report the derivation of a cloned cell line (R278.5) from a rhesus monkey blastocyst that remains undifferentiated in continuous passage for >1 year, maintains a normal XY karyotype, and expresses the cell surface markers (alkaline phosphatase, stage-specific embryonic antigen 3, stage-specific embryonic antigen 4, TRA-1-60, and TRA-1-81) that are characteristic of human embryonal carcinoma cells. R278.5 cells remain undifferentiated when grown on mouse embryonic fibroblast feeder layers but differentiate or die in the absence of fibroblasts, despite the presence of recombinant human leukemia inhibitory factor. R278.5 cells allowed to differentiate in vitro secrete bioactive chorionic gonadotropin into the medium, express chorionic gonadotropin α- and β-subunit mRNAs, and express α-fetoprotein mRNA, indicating trophoblast and endoderm differentiation. When injected into severe combined immunodeficient mice, R278.5 cells consistently differentiate into derivatives of all three embryonic germ layers. These results define R278.5 cells as an embryonic stem cell line, to our knowledge, the first to be derived from any primate species.