Generation of T cells from human embryonic stem cell-derived hematopoietic zones.
Human embryonic stem cells (hESC) are pluripotent stem cells. A major challenge in the field of hESC is the establishment of specific differentiation protocols that drives hESC down a particular lineage fate. So far, attempts to generate T cells from hESC in vitro were unsuccessful. In this study, we show that T cells can be generated in vitro from hESC-derived hematopoietic precursor cells present in hematopoietic zones (HZs). These zones are morphologically similar to blood islands during embryonic development, and are formed when hESC are cultured on OP9 stromal cells. Upon subsequent transfer of these HZs on OP9 cells expressing high levels of Delta-like 1 and in the presence of growth factors, cells expand and differentiate to T cells. Furthermore, we show that T cells derive exclusively from a CD34(high)CD43(low) population, further substantiating the notion that hESC-derived CD34(high)CD43(low) cells are formed in HZs and are the only population containing multipotent hematopoietic precursor cells. Differentiation to T cells sequentially passes through the physiological intermediates: CD34(+)CD7(+) T/NK committed, CD7(+)CD4(+)CD8(-) immature single positive, CD4(+)CD8(+) double positive, and finally CD3(+)CD1(-)CD27(+) mature T cell stages. TCRalphabeta(+) and TCRgammadelta(+) T cells are generated. Mature T cells are polyclonal, proliferate, and secrete cytokines in response to mitogens. This protocol for the de novo generation of T cells from hESC could be clinically and scientifically relevant.
Timmermans
Frank
F
Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
Velghe
Imke
I
Vanwalleghem
Lieve
L
De Smedt
Magda
M
Van Coppernolle
Stefanie
S
Taghon
Tom
T
Moore
Harry D
HD
Leclercq
Georges
G
Langerak
Anton W
AW
Kerre
Tessa
T
Plum
Jean
J
Vandekerckhove
Bart
B
D015703:Antigens, CD / Q000032:analysis; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology*; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D017154:Stromal Cells; D013601:T-Lymphocytes / Q000166:cytology*
D015703:Antigens, CD