HES-1 preserves purified hematopoietic stem cells ex vivo and accumulates side population cells in vivo.
Mouse long-term hematopoietic reconstituting cells exist in the c-Kit+Sca-1+Lin- (KSL) cell population; among them, CD34(low/-) cells represent the most highly purified population of hematopoietic stem cells in the adult bone marrow. Here, we demonstrate that retrovirus-mediated transduction of CD34(low/-)c-Kit+Sca-1+Lin- (34-KSL) cells with the HES-1 gene, which encodes a basic helix-loop-helix transcription factor functioning downstream of the Notch receptor, and is a key molecule for the growth phase of neural stem cells in the embryo, preserves the long-term reconstituting activity of these cells in vitro. We also show that cells derived from the HES-1-transduced 34-KSL population produce progenies characterized by negative Hoechst dye staining, which defines the side population, and by CD34(low/-) profile in the bone marrow KSL population in each recipient mouse at ratios 3.5- and 7.8-fold those produced by nontransduced 34-KSL-derived competitor cells. We conclude that HES-1 preserves the long-term reconstituting hematopoietic activity of 34-KSL stem cells ex vivo. Up-regulation of HES-1 protein in the 34-KSL population before unnecessary cell division, that is, without retrovirus transduction, may represent a potent approach to absolute expansion of hematopoietic stem cells.
Kunisato
Atsushi
A
Departments of Hematology and Oncology, Graduate School of Medicine, and Cell Therapy and Transplantation Medicine, University of Tokyo Hospital, University of Tokyo, Japan.
Chiba
Shigeru
S
Nakagami-Yamaguchi
Etsuko
E
Kumano
Keiki
K
Saito
Toshiki
T
Masuda
Shigeo
S
Yamaguchi
Tomoyuki
T
Osawa
Masatake
M
Kageyama
Ryoichiro
R
Nakauchi
Hiromitsu
H
Nishikawa
Mitsuo
M
Hirai
Hisamaru
H
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins; D019070:Cell Lineage; D002470:Cell Survival; D002478:Cells, Cultured / Q000166:cytology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000145:classification / Q000166:cytology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000502:physiology*; D008164:Luminescent Proteins / Q000032:analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D010641:Phenotype; D011828:Radiation Chimera; D011993:Recombinant Fusion Proteins / Q000502:physiology; D000072056:Transcription Factor HES-1; D014161:Transduction, Genetic
D051792:Basic Helix-Loop-Helix Proteins; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000072056:Transcription Factor HES-1; C489734:Hes1 protein, mouse