In vitro human T cell development directed by notch-ligand interactions.
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.
Awong
Génève
G
Department of Immunology, University of Toronto,Sunnybrook Research Institute, Toronto, Ontario, Canada.
La Motte-Mohs
Ross N
RN
Zúñiga-Pflücker
Juan Carlos
JC
D002460:Cell Line; D018920:Coculture Techniques; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D008024:Ligands; D051880:Receptors, Notch / Q000378:metabolism*; D013601:T-Lymphocytes / Q000166:cytology*
D008024:Ligands; D051880:Receptors, Notch