Diabetes impairs mobilization of mouse bone marrow-derived Lin(-)/VEGF-R2(+) progenitor cells

Abstract

Endothelial progenitor cells circulating in the peripheral blood (PB) contribute to vascular repair. This study aimed to evaluate the potential of a 'cocktail' consisting of erythropoietin, granulocyte colony-stimulating factor and tetrahydrobiopterin to mobilize hematopoietic lineage negative/vascular endothelial growth factor receptor 2 positive (Lin(-)/VEGF-R2(+)) cells from the bone marrow (BM) to PB in non-diabetic and diabetic mice. Diabetes was induced in mice by intraperitoneal injection of streptozotocin. Diabetic mice were studied after 16weeks of hyperglycemia. Half the mice in each group (non-diabetic and diabetic) received daily intraperitoneal injections of the cocktail for 6 consecutive days while the other half received vehicle buffer. Mobilization of Lin(-)/VEGF-R2(+) cells, which were expanded in MCP301 medium, was evaluated after isolating them from BM and PB and their phenotypic and morphological properties were studied. We found that 16weeks of diabetes affected neither the total number of BM mononucleated cells nor the number of Lin(-)/VEGF-R2(+) cells in BM compared with non-diabetic controls. In non-diabetic mice, cocktail treatment resulted in a significant decrease in BM Lin(-)/VEGF-R2(+) cells, paralleled by a significant increase of these cells in PB. Such changes in the number of Lin(-)/VEGF-R2(+) cells in BM and PB after the cocktail treatment were less marked in diabetic mice. In vitro studies of BM Lin(-)/VEGF-R2(+) cells from diabetic and non-diabetic mice did not reveal any differences in either phenotypes or colony forming potential. These findings indicate that diabetes impairs the mobilization of Lin(-)/VEGF-R2(+) cells from BM to PB. Impaired mobilization of BM Lin(-)/VEGF-R2(+) cells soon after the onset of diabetes may contribute to complications such as diabetic retinopathy.

Keywords: ARVO; Association for research in vision and ophthalmology; Australian national health and medical research council; BH4; BM; BRB; Biotinylated Ulex europaeus agglutinin I; Blood-retinal barrier; Bone marrow; CNTX; CTX; Control mice treated with the drug cocktail; Control mice without drug cocktail treatment; DNTX; DTX; Diabetes; Diabetic mice treated with the drug cocktail; Diabetic mice without drug cocktail treatment; Diabetic retinopathy; EPCs; EPO; Endothelial progenitor cells; Erythropoietin; FFA; Fundus fluorescein angiography; G-CSF; Granulocyte colony-stimulating factor; Hematopoietic lineage markers; Intraperitoneal; Lin; NHMRC; PB; Peripheral blood; SD; SDF-1; STZ; Standard deviation; Stem cell mobilization; Streptozotocin; Stromal derived factor-1; Tetrahydrobiopterin; UEA-I; VEGF; VEGF-R2; Vascular endothelial growth factor; Vascular endothelial growth factor receptor 2; WBCs; White blood cells; i.p.