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In vitro human T cell development directed by notch-ligand interactions

Génève Awong1, Ross N La Motte-Mohs, Juan Carlos Zúñiga-Pflücker

  • 1Department of Immunology, University of Toronto,Sunnybrook Research Institute, Toronto, Ontario, Canada.

Methods in Molecular Biology (Clifton, N.J.)
|March 29, 2008
PubMed

Traditionally, the study of human T cell development has relied on the availability of human and mouse thymic tissue. In this chapter, we outline a simple in vitro protocol for generating large numbers of human T-lineage cells from umbilical cord blood (CB)- derived hematopoietic stem cells (HSCs) using a bone marrow stromal cell line. This protocol is broken into three major steps: (1) the maintenance of a working stock of OP9 bone marrow stromal cells expressing the Notch receptor ligand Delta-like 1 (OP9- DL1), (2) the purification of human HSCs from umbilical CB, and (3) the initiation and maintenance/expansion of OP9-DL1 cocultures over time (see Fig. 1). The use of this system opens avenues for basic research as it equips us with a simple in vitro method for studying human T cell development.

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