Computational protocol: Heparan Sulfate Inhibits Hematopoietic Stem and Progenitor Cell Migration and Engraftment in Mucopolysaccharidosis I*

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Protocol publication

[…] All flow cytometry was carried out on a BD FacsCANTO II flow cytometer, using FacsDiva (BD), FlowJo (TreeStar) or Weasel (WEHI Institute) software for analysis. ToPro3 (Invitrogen) or 7AAD (BD Biosciences) were used as viability stains. For the identification of hematopoietic stem and progenitor populations, the following antibodies were used: Rat αMouse C-Kit FITC; Lineage Mixture (CD3e, CD11b, CD45R, TER-119, Ly6G, Ly6C) APC; Rat αMouse Sca-1 PE; Rat αMouse CD45 APC-Cy7 (all BD Biosciences). For the identification of mature hematopoietic lineages, the following antibodies were used: Rat αMouse CD11b PE-Cy7; Rat αMouse CD19; Rat αMouse CD3 PerCPCy5.5 (all BD Biosciences). For measurement of donor chimerism: Mouse αMouse CD45.1 PE; Mouse αMouse CD45.2 (BD Biosciences). For the characterization of MSCs: Rat αMouse C-Kit FITC; Rat αMouse CD11b PE-Cy7; Rat αMouse Sca-1 PE; Rat αMouse CD45 APC-Cy7; Rat αMouse CD44 PerCP Cy5.5; Rat αMouse Sca-1 PE-Cy7 (BD Biosciences), and Rat αMouse CD105 FITC (R&D Systems). For the analysis of HS, the following phage display scFv antibodies were used: HS4C3, RB4EA12 and MPB49 as described previously (, ). […]

Pipeline specifications

Software tools FlowJo, Weasel
Application Flow cytometry
Organisms Mus musculus, Homo sapiens
Diseases Mucopolysaccharidosis I, Mucopolysaccharidosis III, Lysosomal Storage Diseases, Neurodegenerative Diseases, Genetic Diseases, Inborn
Chemicals Heparitin Sulfate