Isolation of hematopoietic stem cells from mouse embryonic stem cells.

Details

Serval ID
serval:BIB_560B26A44773
Type
Article: article from journal or magazin.
Collection
Publications
Title
Isolation of hematopoietic stem cells from mouse embryonic stem cells.
Journal
Current protocols in stem cell biology
Author(s)
McKinney-Freeman S.L., Naveiras O., Daley G.Q.
ISSN
1938-8969 (Electronic)
ISSN-L
1938-8969
Publication state
Published
Issued date
02/2008
Peer-reviewed
Oui
Volume
Chapter 1
Pages
Unit 1F.3
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
This unit describes a protocol for the isolation of cells from murine embryonic stem cells with hematopoietic stem cell activity, defined by the ability to reconstitute, long term, multiple lineages of the hematopoietic system of lethally irradiated mice. The protocol subjects hematopoietic progenitors specified in differentiating embryoid bodies to ectopic HoxB4 expression (delivered via retroviral infection), followed by coculture and expansion on OP9 stromal cells in the presence of hematopoietic cytokines for 10 days. The protocol results in the generation of hundreds of millions of cells that can rescue mice from lethal irradiation. Although little is known about the phenotype and frequency of the actual hematopoietic stem cell-like cell within the population of cells generated by this protocol, the protocol establishes a system in which these cells can be further studied and the results ultimately translated to the human system.
Keywords
Animals, Cell Differentiation, Cell Proliferation, Cell Separation/methods, Cells, Cultured, Embryonic Stem Cells/cytology, Green Fluorescent Proteins/metabolism, Hematopoietic Stem Cells/cytology, Homeodomain Proteins/metabolism, Mice, Retroviridae, Stem Cell Transplantation, Stromal Cells, Transcription Factors/metabolism
Pubmed
Create date
11/10/2022 0:06
Last modification date
11/10/2022 6:39
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