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Prepublished online as a Blood First Edition Paper on May 8, 2003; DOI 10.1182/blood-2003-01-0224.
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Blood, 1 September 2003, Vol. 102, No. 5, pp. 1649-1653
HEMATOPOIESIS
In vitro generation of T lymphocytes from embryonic stem cellderived prehematopoietic progenitors
Renée F. de Pooter,
Sarah K. Cho,
James R. Carlyle, and
Juan Carlos Zúñiga-Pflücker
From the Department of Immunology, University of Toronto, ON,
Canada.
Embryonic stem (ES) cells can differentiate into most blood cells in vitro,
providing a powerful model system to study hematopoiesis. However, ES
cellderived T lymphocytes have not been generated in vitro, and it was
unresolved whether such potential is absent or merely difficult to isolate.
Because the latter case might result from rapid commitment to nonT-cell
fates, we isolated ES cellderived prehematopoietic precursors for
reconstitution of fetal thymic organ cultures. We found a transient
Flk1+CD45 subset of these precursors generated T
lymphocytes in vitro, and the use of reaggregate thymic organ cultures greatly
enhanced reconstitution frequency. These findings reveal that ES cells can
exhibit in vitro T-cell potential, but this is restricted to early stages of
ES cell differentiation. Moreover, the results support the notion that the
thymic microenvironment can induce T-cell differentiation from a subset of
prehematopoietic progenitors and suggest deficient migration into intact thymi
hindered previous attempts to generate T cells in vitro from ES
cellderived progenitors. These findings demonstrate that a defined
subset of ES cells has the potential to generate T cells in vitro and could
contribute to greater understanding of the molecular events of hematopoietic
induction and T-cell lineage commitment.

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