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Isolation of a primate embryonic stem cell line
J A Thomson1, J Kalishman, T G Golos
1Wisconsin Regional Primate Research Center, University of Wisconsin, Madison 53715-1299, USA.
Summary
Researchers derived a new primate embryonic stem cell line from rhesus monkeys. This cell line, R278.5, maintains pluripotency and can differentiate into all three embryonic germ layers.
Area of Science:
- Stem Cell Biology
- Primate Embryology
- Developmental Biology
Background:
- Embryonic stem cells (ESCs) are crucial for developmental biology research due to their pluripotency.
- Deriving and characterizing ESCs from various species is essential for comparative studies and understanding early development.
- Primate ESCs are particularly valuable for modeling human development and disease.
Purpose of the Study:
- To derive and characterize a novel embryonic stem cell line from a rhesus monkey blastocyst.
- To assess the pluripotency and differentiation potential of the derived cell line.
- To establish the first primate ESC line for broader research applications.
Main Methods:
- Derivation of a cloned cell line (R278.5) from a rhesus monkey blastocyst.
- Continuous in vitro culture and passage for over one year.
- Karyotyping and analysis of characteristic ESC surface markers (alkaline phosphatase, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81).
- In vitro differentiation assays with and without feeder layers and leukemia inhibitory factor.
- In vivo differentiation studies following injection into severe combined immunodeficient (SCID) mice.
Main Results:
- The R278.5 cell line maintained undifferentiated status for over a year with a normal XY karyotype.
- Cells expressed key markers characteristic of human embryonal carcinoma cells and ESCs.
- R278.5 cells required feeder layers for maintaining pluripotency, differentiating or dying without them.
- In vitro differentiation induced secretion of chorionic gonadotropin and expression of alpha-fetoprotein mRNA.
- In vivo studies demonstrated differentiation into derivatives of all three embryonic germ layers.
Conclusions:
- The R278.5 cell line represents a stable, pluripotent embryonic stem cell line derived from a primate.
- This marks the first derivation of an ESC line from any primate species.
- The R278.5 line provides a valuable tool for studying primate development, stem cell biology, and disease modeling.