由Notch配体相互作用介导的体外人T细胞发育

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

作者信息

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

机构信息

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

出版信息

Methods Mol Biol. 2008;430:135-42. doi: 10.1007/978-1-59745-182-6_9.

DOI:10.1007/978-1-59745-182-6_9

PMID:18370296

Abstract

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.

摘要

传统上，人类T细胞发育的研究依赖于人类和小鼠胸腺组织的可得性。在本章中，我们概述了一种简单的体外实验方案，该方案使用骨髓基质细胞系从脐带血（CB）来源的造血干细胞（HSC）中生成大量人类T系细胞。该方案分为三个主要步骤：（1）维持表达Notch受体配体Delta样1的OP9骨髓基质细胞（OP9-DL1）的工作储备；（2）从脐带血中纯化人类造血干细胞；（3）随着时间的推移启动并维持/扩大OP9-DL1共培养体系（见图1）。该系统的使用为基础研究开辟了道路，因为它为我们提供了一种研究人类T细胞发育的简单体外方法。