Generation of immunogenic dendritic cells from human embryonic stem cells without serum and feeder cells.
OBJECTIVE
Dendritic cell (DC)-based vaccines have a potential utility for use in the treatment of malignancy. Human embryonic stem cells (hESCs) may provide a more cost-effective and reliable source of DCs for immunotherapy purposes, providing on-demand access for patients.


METHODS
We developed a protocol to generate DCs from hESCs in vitro in the absence of serum and feeder cells. This protocol uses growth factors bone morphogenetic protein-4, granulocyte macrophage-colony stimulating factor (GM-CSF), stem cell factor and VEGF in serum-free media to generate hESC-derived monocytic cells. These cells are further differentiated to hESC-derived immature DCs with GM-CSF and IL-4, and matured to hESC-derived mature DCs with a maturation cocktail consisting of GM-CSF, TNF-alpha, IL-1beta, IFN-gamma and PGE2.


RESULTS
This study demonstrates the applicability of our defined differentiation process in generating functional hESC-derived DCs from multiple hESC lines. We show that hESC-derived immature DCs phagocytose, process, and present antigen upon maturation. hESC-derived mature DCs express the maturation marker CD83, produce Th1-directing cytokine IL-12p70, migrate in response to chemokine, and activate both viral and tumor antigen-specific T-cell responses.


CONCLUSIONS
We developed a chemically defined system to generate unlimited numbers of DCs from hESCs. Our results demonstrate that hESC-derived DCs generated from this process are immunogenic and have the potential to be used for DC immunotherapy.
Tseng
Su-Yi
SY
Geron Corporation, 230 Constitution Drive, Menlo Park, CA 94025, USA. stseng@geron.com
Nishimoto
Kevin P
KP
Silk
Kathryn M
KM
Majumdar
Anish S
AS
Dawes
Glenn N
GN
Waldmann
Herman
H
Fairchild
Paul J
PJ
Lebkowski
Jane S
JS
Reddy
Anita
A
D001709:Biotechnology / Q000379:methods*; D055415:Bone Morphogenetic Protein 4; D002454:Cell Differentiation / Q000502:physiology*; D003713:Dendritic Cells / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D019089:Stem Cell Factor; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D019098:Telomerase / Q000378:metabolism; D014612:Vaccines / Q000096:biosynthesis*; D042461:Vascular Endothelial Growth Factor A
C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D019089:Stem Cell Factor; C467484:VEGFA protein, human; D014612:Vaccines; D042461:Vascular Endothelial Growth Factor A; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; C509186:TERT protein, human; D019098:Telomerase