Abstract
The unique properties of the hematopoietic progenitor cell (HPC) lines offer a valuable tool for directly addressing basic properties of HSCs, such as signal transduction, cytokine/growth factor response, lineage commitment and adhesion to support cells. Thus, characterization of these cell lines might also be useful for elucidating mechanisms involved in embryonic development of the hematopoietic system. An experimental system has been developed that makes it possible to address both of these aspects of the hematopoietic system. This system is based on the generation of immortalized HPC lines from in vitro differentiation of mouse embryonic stem (ES) cells genetically modified to over-express the LIM-homeobox gene Lhx2 . Moreover, Lhx2 appears to play an important role in the establishment of the hematopoietic system during embryonic development, as Lhx2 - / - mice die in utero due to severe anemia. This chapter describes a detailed experimental approach for generating HPC lines from genetically modified ES cells that have been allowed to differentiate in vitro . The estimated time to generate HPC lines, including genetic modification of ES cells, is approximately 2 months. The general outline of the experimental approach and the estimated time for each step of the procedure are illustrated in this chapter.
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