Highly efficient ex vivo expansion of human hematopoietic stem cells using Delta1-Fc chimeric protein.
Ex vivo expansion of hematopoietic stem cells (HSCs) has been explored in the fields of stem cell biology, gene therapy, and clinical transplantation. Here, we demonstrate efficient ex vivo expansion of HSCs measured by long-term severe combined immunodeficient (SCID) repopulating cells (SRCs) from human cord blood CD133-sorted cells using a soluble form of Delta1. After a 3-week culture on immobilized Delta1 supplemented with stem cell factor, thrombopoietin, Flt-3 ligand, interleukin (IL)-3, and IL-6/soluble IL-6 receptor chimeric protein (FP6) in a serum- and stromal cell-free condition, we achieved approximately sixfold expansion of SRCs when evaluated by limiting dilution/transplantation assays. The maintenance of full multipotency and self-renewal capacity during culture was confirmed by transplantation to nonobese diabetic/SCID/gammac(null) mice, which showed myeloid, B, T, and natural killer cells as well as CD133(+)CD34(+) cells, and hematopoietic reconstitution in the secondary recipients. Interestingly, the CD133-sorted cells contained approximately 4.5 times more SRCs than the CD34-sorted cells. The present study provides a promising method to expand HSCs and encourages future trials on clinical transplantation.
Suzuki
Takahiro
T
Department of Regeneration Medicine for Hematopoiesis, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.
Yokoyama
Yasuhisa
Y
Kumano
Keiki
K
Takanashi
Minoko
M
Kozuma
Shiro
S
Takato
Tsuyoshi
T
Nakahata
Tatsutoshi
T
Nishikawa
Mitsuo
M
Sakano
Seiji
S
Kurokawa
Mineo
M
Ogawa
Seishi
S
Chiba
Shigeru
S
D000071916:AC133 Antigen; D051997:ADP-ribosyl Cyclase 1 / Q000032:analysis; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D050822:Cytokine Receptor gp130 / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D006023:Glycoproteins / Q000032:analysis; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D006801:Humans; D018189:Immunomagnetic Separation; D007377:Interleukin-3 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D010455:Peptides / Q000032:analysis; D011961:Receptors, Fc / Q000235:genetics / Q000378:metabolism*; D018123:Receptors, Interleukin / Q000378:metabolism; D019947:Receptors, Interleukin-6; D051880:Receptors, Notch / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D006023:Glycoproteins; D007377:Interleukin-3; D015850:Interleukin-6; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D011961:Receptors, Fc; D018123:Receptors, Interleukin; D019947:Receptors, Interleukin-6; D051880:Receptors, Notch; D011993:Recombinant Fusion Proteins; C064045:delta protein; C104543:interleukin 6-interleukin 6 receptor fusion protein, recombinant; D050822:Cytokine Receptor gp130; D051997:ADP-ribosyl Cyclase 1