Abstract
The earliest lymphoid precursor in the adult mouse thymus, the “low CD4 precursor,” was found to be able to produce T cells, B cells, NK cells, and dendritic cells (DC) upon adoptive transfer (1-3). This precursor population represents only 0.03%-0.05% of total thymocytes and expresses low levels of CD4 and Thy-1, and positive for c-kit and CD44. The principle for isolating this minute precursor population is to enrich maximally for the population prior to fluorescence activated cell separation in order to reduce the cost and to maximize purity. A combination of density centrifugation and immunomagnetic bead depletion successfully removes mature and immature thymocytes and non-T-lineage cells. Note, it is important to deplete non-T-lineage cells, including erythrocytes, macrophages, and DC, which may otherwise contaminate the precursor preparation.
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References
Wu, L., Antica, M., Johnson, G. R., Scollay, R., and Shortman, K. (1991) Developmental potential of the earliest precursor cells from the adult mouse thymus. J. Exp. Med. 174, 1617–1627.
Matsuzaki, Y., Gyotoku, J., Ogawa, M., Nishikawa, S., Katsura, Y., Gachelin, G., and Nakauchi, H. (1993) Characterization of c-kit positive intrathymic stem cells that are restricted to lymphoid differentiation. J. Exp. Med. 178, 1283–1291.
Ardavin, C., Wu, L., Li, C., and Shortman, K. (1993) Thymic dendritic cells and T cells develop simultaneously within the thymus from a common precursor population. Nature 362, 761–763.
Steinman, R. M. (1991) The dendritic cell system and its role in immunogenicity. Ann. Rev. Immunol. 9, 271–296.
Inaba, K., Inaba, M., Romani, N., Aya, H., Deguchi, M., Ikehara, S., Muramatsu, S., and Steinman, R. M. (1992) Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J. Exp. Med. 176, 1693–1702.
Caux, C. and Banchereau, J. (1996) In vitro regulation of dendritic cell development and function, in Blood cell biochemistry, Vol. 7, Hemopoietic growth factors and their receptors (Whetton, A. and Gordons, J., eds.), Plenum Press, London, pp. 263–301.
Saunders, D., Lucas, K., Ismaili, J., Wu, L., Maraskovsky, E., Dunn, A., and Shortman, K. (1996) Dendritic cell development in culture from thymic precursor cells in the absence of granulocyte/macrophage colony-stimulating factor. J. Exp. Med. 184, 2185–2196.
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Lucas, K., Saunders, D., Wu, L. (2001). Derivation of Murine Dendritic Cells from Thymic Precursors. In: Robinson, S.P., Stagg, A.J., Knight, S.C. (eds) Dendritic Cell Protocols. Methods in Molecular Medicine™, vol 64. Humana Press. https://doi.org/10.1385/1-59259-150-7:207
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DOI: https://doi.org/10.1385/1-59259-150-7:207
Publisher Name: Humana Press
Print ISBN: 978-0-89603-584-3
Online ISBN: 978-1-59259-150-3
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