| RRC ID |
62185
|
| Author |
Awong G, La Motte-Mohs RN, Zúñiga-Pflücker JC.
|
| Title |
In vitro human T cell development directed by notch-ligand interactions.
|
| Journal |
Methods Mol Biol
|
| 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.
|
| Volume |
430
|
| Pages |
135-42
|
| Published |
2008-1-1
|
| DOI |
10.1007/978-1-59745-182-6_9
|
| PMID |
18370296
|
| MeSH |
Cell Line
Coculture Techniques
Flow Cytometry
Hematopoietic Stem Cells / cytology
Humans
Ligands
Receptors, Notch / metabolism*
T-Lymphocytes / cytology*
|
| Resource |
| Human and Animal Cells |
OP9(RCB1124) |
