Application

Cell line for efficient in vitro differentiation of rhesus macaque-specific T cells from hematopoietic stem and progenitor cells.

Key Benefits

• Replaces need for primary rhesus macaque thymus tissue.
• Improves differentiation efficiency and reduces media requirements compared to existing methods.
• Yields CD4+ and CD8+ T cells, double-positive and double-negative thymocytes.
• Yields TCR selected CD4 and CD8 T cells.

Market Summary

Nonhuman primates serve as critical models for evaluating human disease pathogenesis and therapeutic interventions. In vitro systems for producing nonhuman primate cell types can enable physiologically relevant preclinical studies at scale. Generating T cells in particular can enable immune system disease modeling and therapeutic testing relevant to cancer, autoimmune, infectious, and inflammatory diseases. However, differentiating nonhuman primate T cells from hematopoietic stem cells is resource-intensive, inefficient, and unscalable. Importantly, generating TCR selected T cells using Non-Animal Methods (NAMs) in NHPs have not been achieved.

Technical Summary

To differentiate hematopoietic or induced pluripotent stem cells into T cells, existing approaches require coculturing stem cells with primary thymic tissue from non-human primates. To enable more cost-effective and efficient rhesus macaque-specific T cell generation, and develop NHP-specific NAMs, Emory researchers generated a mouse stromal cell (MS5) line with stable expression of the rhesus DLL1 gene (Delta-like ligand 1). The cell line also features puromycin resistance to enable purification and a fluorescent marker (eGFP) for visualization. When cocultured with this MS5-rhesusDLL1 cell line, stem cells reliably and efficiently differentiate into cell types across the T cell lineage—including pro-T thymocytes, CD4+CD3- immature single positive, CD4+CD8+ double positive early thymocytes, CD3+ T cells including CD4+ and CD8+ T cell subsets—within 2 weeks. With robust, fast differentiation demonstrated in 3D stem cell models, this MS5-rhesusDLL1 cell line can significantly advance preclinical research and therapeutic testing scale for nonhuman primate T cell lineages including generation of TCR selected NHP T cells.

Development Stage

This cell line has been generated, tested in 3D cell coculture systems, and evaluated for differentiation efficiency and efficacy (Wilde et al., 2025).

Publication

Wilde, C., Anwar, S., Yau, Y.-T., Badve, S., Polar, Y.G., Roback, J.D., Amara, R.R., Johnson, R.P., and Rahman, S.A. (2025). A Thymus-Independent Artificial Organoid System Supports Complete Thymopoiesis from Rhesus Macaque-Derived Hematopoietic Stem and Progenitor Cells. Biomedicines 13, 2692.