Springer Protocols: Abstract: In Vitro Human T Cell Development Directed by Notch

In Vitro Human T Cell Development Directed by Notch–Ligand Interactions

By: Génève Awong³, Ross N. La Motte-Mohs³, Juan Carlos Zúñiga-Pflücker³

Abstract

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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.

Affiliation(s): (3) Department of Immunology, University of Toronto,Sunnybrook Research Institute, Toronto, Ontario, Canada

Book Title: Hematopoietic Stem Cell Protocols

Series: Methods in Molecular Biology | Volume: 430 | Pub. Date: Jan-01-2008 | Page Range: 135-142 | DOI: 10.1007/978-1-59745-182-6_9

Subject: Cell Biology

Key Words: T cell development - lymphopoiesis - umbilical cord blood - Notch - Delta-like 1 - CD34 - hematopoietic stem cells - stromal cells - IL-7

References

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