Highly efficient ex vivo expansion of human hematopoietic stem cells using Delta1-Fc chimeric protein

Takahiro Suzuki; Yasuhisa Yokoyama; Keiki Kumano; Minoko Takanashi; Shiro Kozuma; Tsuyoshi Takato; Tatsutoshi Nakahata; Mitsuo Nishikawa; Seiji Sakano; Mineo Kurokawa; Seishi Ogawa; Shigeru Chiba

doi:10.1634/stemcells.2006-0258

Highly efficient ex vivo expansion of human hematopoietic stem cells using Delta1-Fc chimeric protein

Stem Cells. 2006 Nov;24(11):2456-65. doi: 10.1634/stemcells.2006-0258. Epub 2006 Jul 20.

Authors

Takahiro Suzuki¹, Yasuhisa Yokoyama, Keiki Kumano, Minoko Takanashi, Shiro Kozuma, Tsuyoshi Takato, Tatsutoshi Nakahata, Mitsuo Nishikawa, Seiji Sakano, Mineo Kurokawa, Seishi Ogawa, Shigeru Chiba

Affiliation

¹ Department of Regeneration Medicine for Hematopoiesis, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.

PMID: 16857897
DOI: 10.1634/stemcells.2006-0258

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

AC133 Antigen
ADP-ribosyl Cyclase 1 / analysis
Animals
Antigens, CD / analysis
Antigens, CD34 / analysis
Cell Differentiation / drug effects
Cell Lineage
Cell Proliferation* / drug effects
Cells, Cultured
Cytokine Receptor gp130 / metabolism
Fetal Blood / cytology
Fetal Blood / immunology
Fetal Blood / metabolism
Glycoproteins / analysis
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells* / drug effects
Hematopoietic Stem Cells* / immunology
Hematopoietic Stem Cells* / metabolism
Humans
Immunomagnetic Separation
Interleukin-3 / metabolism
Interleukin-6 / metabolism
Intracellular Signaling Peptides and Proteins
Membrane Proteins / genetics
Membrane Proteins / metabolism*
Mice
Mice, Inbred NOD
Mice, Knockout
Mice, SCID
Peptides / analysis
Receptors, Fc / genetics
Receptors, Fc / metabolism*
Receptors, Interleukin / metabolism
Receptors, Interleukin-6
Receptors, Notch / metabolism
Recombinant Fusion Proteins / metabolism

Substances

AC133 Antigen
Antigens, CD
Antigens, CD34
Glycoproteins
Interleukin-3
Interleukin-6
Intracellular Signaling Peptides and Proteins
Membrane Proteins
PROM1 protein, human
Peptides
Prom1 protein, mouse
Receptors, Fc
Receptors, Interleukin
Receptors, Interleukin-6
Receptors, Notch
Recombinant Fusion Proteins
delta protein
interleukin 6-interleukin 6 receptor fusion protein, recombinant
Cytokine Receptor gp130
ADP-ribosyl Cyclase 1