title: Selection based on CD133 and high aldehyde dehydrogenase activity isolates long-term reconstituting human hematopoietic stem cells.
abstract: The development of novel cell-based therapies requires understanding of distinct human hematopoietic stem and progenitor cell populations. We recently isolated reconstituting hematopoietic stem cells (HSCs) by lineage depletion and purification based on high aldehyde dehydrogenase activity (ALDH(hi)Lin- cells). Here, we further dissected the ALDH(hi)-Lin- population by selection for CD133, a surface molecule expressed on progenitors from hematopoietic, endothelial, and neural lineages. ALDH(hi)CD133+Lin- cells were primarily CD34+, but also included CD34-CD38-CD133+ cells, a phenotype previously associated with repopulating function. Both ALDH(hi)CD133-Lin- and ALDH(hi)CD133+Lin- cells demonstrated distinct clonogenic progenitor function in vitro, whereas only the ALDH(hi)CD133+Lin- population seeded the murine bone marrow 48 hours after transplantation. Significant human cell repopulation was observed only in NOD/SCID and NOD/SCID beta2M-null mice that received transplants of ALDH(hi)CD133+Lin- cells. Limiting dilution analysis demonstrated a 10-fold increase in the frequency of NOD/SCID repopulating cells compared with CD133+Lin- cells, suggesting that high ALDH activity further purified cells with repopulating function. Transplanted ALDH(hi)CD133+Lin- cells also maintained primitive hematopoietic phenotypes (CD34+CD38-) and demonstrated enhanced repopulating function in recipients of serial, secondary transplants. Cell selection based on ALDH activity and CD133 expression provides a novel purification of HSCs with long-term repopulating function and may be considered an alternative to CD34 cell selection for stem cell therapies.
authors.affiliation: Department of Internal Medicine, Division of Oncology, Hematopoietic Development and Malignancy Group, Washington University School of Medicine, Southwest Tower, Rm 644, 4940 Parkview Pl, Campus Box 8007, St Louis, MO 63110, USA. dhess@im.wustl.edu
authors.forename: David A
authors.initials: DA
authors.lastname: Hess
authors.forename: Louisa
authors.initials: L
authors.lastname: Wirthlin
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Craft
authors.forename: Phillip E
authors.initials: PE
authors.lastname: Herrbrich
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Hohm
authors.forename: Ryan
authors.initials: R
authors.lastname: Lahey
authors.forename: William C
authors.initials: WC
authors.lastname: Eades
authors.forename: Michael H
authors.initials: MH
authors.lastname: Creer
authors.forename: Jan A
authors.initials: JA
authors.lastname: Nolta
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D050822:Cytokine Receptor gp130 / Q000378:metabolism*; D006085:Graft Survival / Q000502:physiology*; D006410:Hematopoiesis; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D018183:Transplantation Chimera / Q000502:physiology
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; C494199:IL6ST protein, human; D050822:Cytokine Receptor gp130; D000444:Aldehyde Dehydrogenase