title: High Aldehyde Dehydrogenase Activity Identifies a Subset of Human Mesenchymal Stromal Cells with Vascular Regenerative Potential.
abstract: During culture expansion, multipotent mesenchymal stromal cells (MSCs) differentially express aldehyde dehydrogenase (ALDH), an intracellular detoxification enzyme that protects long-lived cells against oxidative stress. Thus, MSC selection based on ALDH-activity may be used to reduce heterogeneity and distinguish MSC subsets with improved regenerative potency. After expansion of human bone marrow-derived MSCs, cell progeny was purified based on low versus high ALDH-activity (ALDHhi) by fluorescence-activated cell sorting, and each subset was compared for multipotent stromal and provascular regenerative functions. Both ALDHl° and ALDHhiMSC subsets demonstrated similar expression of stromal cell (>95% CD73+, CD90+, CD105+) and pericyte (>95% CD146+) surface markers and showed multipotent differentiation into bone, cartilage, and adipose cells in vitro. Conditioned media (CDM) generated by ALDHhiMSCs demonstrated a potent proliferative and prosurvival effect on human microvascular endothelial cells (HMVECs) under serum-free conditions and augmented HMVEC tube-forming capacity in growth factor-reduced matrices. After subcutaneous transplantation within directed in vivo angiogenesis assay implants into immunodeficient mice, ALDHhiMSC or CDM produced by ALDHhiMSC significantly augmented murine vascular cell recruitment and perfused vessel infiltration compared with ALDHl° MSC. Although both subsets demonstrated strikingly similar mRNA expression patterns, quantitative proteomic analyses performed on subset-specific CDM revealed the ALDHhiMSC subset uniquely secreted multiple proangiogenic cytokines (vascular endothelial growth factor beta, platelet derived growth factor alpha, and angiogenin) and actively produced multiple factors with chemoattractant (transforming growth factor-β, C-X-C motif chemokine ligand 1, 2, and 3 (GRO), C-C motif chemokine ligand 5 (RANTES), monocyte chemotactic protein 1 (MCP-1), interleukin [IL]-6, IL-8) and matrix-modifying functions (tissue inhibitor of metalloprotinase 1 & 2 (TIMP1/2)). Collectively, MSCs selected for ALDHhidemonstrated enhanced proangiogenic secretory functions and represent a purified MSC subset amenable for vascular regenerative applications. Stem Cells 2017;35:1542-1553.
authors.affiliation: Krembil Centre for Stem Cell Biology, Molecular Medicine Research Group, Robarts Research Institute, London, Ontario, Canada.|Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, Western University, London, Ontario, Canada.
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Sherman
authors.affiliation: Don Rix Protein Identification Facility, Department of Biochemistry, Schulich School of Medicine and Dentistry, Western University, London, Ontario, Canada.
authors.forename: Miljan
authors.initials: M
authors.lastname: Kuljanin
authors.affiliation: Krembil Centre for Stem Cell Biology, Molecular Medicine Research Group, Robarts Research Institute, London, Ontario, Canada.|Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, Western University, London, Ontario, Canada.
authors.forename: Tyler T
authors.initials: TT
authors.lastname: Cooper
authors.affiliation: Krembil Centre for Stem Cell Biology, Molecular Medicine Research Group, Robarts Research Institute, London, Ontario, Canada.|Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, Western University, London, Ontario, Canada.
authors.forename: David M
authors.initials: DM
authors.lastname: Putman
authors.affiliation: Don Rix Protein Identification Facility, Department of Biochemistry, Schulich School of Medicine and Dentistry, Western University, London, Ontario, Canada.
authors.forename: Gilles A
authors.initials: GA
authors.lastname: Lajoie
authors.affiliation: Krembil Centre for Stem Cell Biology, Molecular Medicine Research Group, Robarts Research Institute, London, Ontario, Canada.|Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, Western University, London, Ontario, Canada.
authors.forename: David A
authors.identifier: 0000-0003-2186-3166
authors.initials: DA
authors.lastname: Hess
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D001807:Blood Vessel Prosthesis; D001808:Blood Vessels / Q000187:drug effects / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D055806:Microvessels / Q000166:cytology; D039902:Multipotent Stem Cells / Q000166:cytology / Q000187:drug effects; D018919:Neovascularization, Physiologic / Q000187:drug effects; D020286:Pericytes / Q000166:cytology / Q000187:drug effects; D020543:Proteome / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012038:Regeneration / Q000187:drug effects; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects
chemical_list: D015415:Biomarkers; D017077:Culture Media, Conditioned; D020543:Proteome; D012333:RNA, Messenger; D000444:Aldehyde Dehydrogenase
keywords: Aldehyde dehydrogenase; Angiogenesis; Multipotent stromal cells; Peripheral artery disease; Proteomics; Transplantation