title: Angiopoietin-like protein 3 promotes preservation of stemness during ex vivo expansion of murine hematopoietic stem cells.
abstract: Allogeneic hematopoietic stem cell (HSC) transplantations from umbilical cord blood or autologous HSCs for gene therapy purposes are hampered by limited number of stem cells. To test the ability to expand HSCs in vitro prior to transplantation, two growth factor cocktails containing stem cell factor, thrombopoietin, fms-related tyrosine kinase-3 ligand (STF) or stem cell factor, thrombopoietin, insulin-like growth factor-2, fibroblast growth factor-1 (STIF) either with or without the addition of angiopoietin-like protein-3 (Angptl3) were used. Culturing HSCs in STF and STIF media for 7 days expanded long-term repopulating stem cells content in vivo by ∼6-fold and ∼10-fold compared to freshly isolated stem cells. Addition of Angptl3 resulted in increased expansion of these populations by ∼17-fold and ∼32-fold, respectively, and was further supported by enforced expression of Angptl3 in HSCs through lentiviral transduction that also promoted HSC expansion. As expansion of highly purified lineage-negative, Sca-1+, c-Kit+ HSCs was less efficient than less pure lineage-negative HSCs, Angptl3 may have a direct effect on HCS but also an indirect effect on accessory cells that support HSC expansion. No evidence for leukemia or toxicity was found during long-term follow up of mice transplanted with ex vivo expanded HSCs or manipulated HSC populations that expressed Angptl3. We conclude that the cytokine combinations used in this study to expand HSCs ex vivo enhances the engraftment in vivo. This has important implications for allogeneic umbilical cord-blood derived HSC transplantations and autologous HSC applications including gene therapy.
authors.affiliation: The Department of Hematology, Erasmus Medical Center, Rotterdam, the Netherlands; The Department of Pediatric Oncology/Hematology, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Elnaz
authors.initials: E
authors.lastname: Farahbakhshian
authors.affiliation: The Department of Hematology, Erasmus Medical Center, Rotterdam, the Netherlands; The Department of Surgery, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Monique M
authors.initials: MM
authors.lastname: Verstegen
authors.affiliation: The Department of Hematology, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Trudi P
authors.initials: TP
authors.lastname: Visser
authors.affiliation: The Department of Cell Biology, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Sima
authors.initials: S
authors.lastname: Kheradmandkia
authors.affiliation: The Department of Pediatric Oncology/Hematology, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Dirk
authors.initials: D
authors.lastname: Geerts
authors.affiliation: The Department of Hematology, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Shazia
authors.initials: S
authors.lastname: Arshad
authors.affiliation: The Department of Hematology, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Noveen
authors.initials: N
authors.lastname: Riaz
authors.affiliation: The Department of Cell Biology, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Frank
authors.initials: F
authors.lastname: Grosveld
authors.affiliation: The Department of Hematology, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Niek P
authors.initials: NP
authors.lastname: van Til
authors.affiliation: The Department of Pediatric Oncology/Hematology, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Jules P P
authors.initials: JP
authors.lastname: Meijerink
mesh_terms: D000091303:Angiopoietin-Like Protein 3; D000075802:Angiopoietin-like Proteins; D042682:Angiopoietins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D000950:Antigens, Ly / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics*; D002478:Cells, Cultured; D005260:Female; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000494:pharmacology; D016086:Lentivirus / Q000235:genetics; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D008807:Mice, Inbred BALB C; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D019089:Stem Cell Factor / Q000494:pharmacology; D013926:Thrombopoietin / Q000494:pharmacology; D014162:Transfection
chemical_list: D000091303:Angiopoietin-Like Protein 3; D000075802:Angiopoietin-like Proteins; D042682:Angiopoietins; C404495:Angptl3 protein, mouse; D000950:Antigens, Ly; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D019089:Stem Cell Factor; D016220:Fibroblast Growth Factor 1; D049452:Green Fluorescent Proteins; D007335:Insulin-Like Growth Factor II; D013926:Thrombopoietin; D019009:Proto-Oncogene Proteins c-kit