title: Bone Marrow Homing Enriches Stem Cells Responsible for Neogenesis of Insulin-Producing Cells, While Radiation Decreases Homing Efficiency.
abstract: Small-sized adult bone marrow cells isolated by counterflow centrifugal elutriation and depleted of lineage markers (Fr25lin(-)) have the capacity to differentiate into insulin-producing cells and stabilize glycemic control. This study assessed competitive migration of syngeneic stem cells to the bone marrow and islets in a murine model of chemical diabetes. VLA-4 is expressed in ∼ 25% of these cells, whereas CXCR4 is not detected, however, it is transcriptionally upregulated (6-fold). The possibility to enrich stem cells by a bone marrow homing (BM-H) functional assay was assessed in sequential transplants. Fr25lin(-) cells labeled with PKH26 were grafted into primary myeloablated recipients, and mitotically quiescent Fr25lin(-)PKH(bright) cells were sorted from the bone marrow after 2 days. The contribution of bone marrow-homed stem cells was remarkably higher in secondary recipients compared to freshly elutriated cells. The therapeutic efficacy was further increased by omission of irradiation in the secondary recipients, showing a 25-fold enrichment of islet-reconstituting cells by the bone marrow homing assay. Donor cells identified by the green fluorescent protein (GFP) and a genomic marker in sex-mismatched transplants upregulated PDX-1 and produced proinsulin, affirming the capacity of BM-H cells to convert in the injured islets. There was no evidence of transcriptional priming of freshly elutriated subsets to express PDX-1, insulin, and other markers of endocrine progenitors, indicating that the bone marrow harbors stem cells with versatile differentiation capacity. Affinity to the bone marrow can be used to enrich stem cells for pancreatic regeneration, and reciprocally, conditioning reduces the competitive incorporation in the injured islets.
authors.affiliation: 1 Krieger Eye Research Laboratory, Schneider Children's Medical Center of Israel , Petach Tikva, Israel .
authors.forename: Nitza
authors.initials: N
authors.lastname: Goldenberg-Cohen
authors.affiliation: 2 Frankel Laboratory, Center for Stem Cell Research, Schneider Children's Medical Center of Israel , Petach Tikva, Israel .
authors.forename: Svetlana
authors.initials: S
authors.lastname: Iskovich
authors.affiliation: 2 Frankel Laboratory, Center for Stem Cell Research, Schneider Children's Medical Center of Israel , Petach Tikva, Israel .
authors.forename: Nadir
authors.initials: N
authors.lastname: Askenasy
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D015153:Blotting, Western; D001853:Bone Marrow / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D016026:Bone Marrow Transplantation / Q000379:methods; D002454:Cell Differentiation; D002465:Cell Movement / Q000528:radiation effects; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000628:therapy; D005260:Female; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D007328:Insulin / Q000097:blood / Q000235:genetics; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism; D039041:Integrin alpha4beta1 / Q000235:genetics; D008297:Male; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics
chemical_list: D001786:Blood Glucose; D018398:Homeodomain Proteins; D007328:Insulin; D039041:Integrin alpha4beta1; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein