[HTML][HTML] Long-term type 1 diabetes influences haematopoietic stem cells by reducing vascular repair potential and increasing inflammatory monocyte generation in a …
S Hazra, YPR Jarajapu, V Stepps, S Caballero… - Diabetologia, 2013 - Springer
S Hazra, YPR Jarajapu, V Stepps, S Caballero, JS Thinschmidt, L Sautina, N Bengtsson…
Diabetologia, 2013•SpringerMethods Wild-type, gp130−/− and GFP chimeric mice were treated with streptozotocin to
induce type 1 diabetes. Bone marrow (BM)-derived cells were used for colony-formation
assay, quantification of side population (SP) cells, examination of gene expression, nitric
oxide measurement and migration studies. Endothelial progenitor cells (EPCs), a population
of vascular precursors derived from HSCs, were compared in diabetic and control mice.
Cytokines were measured in BM supernatant fractions by ELISA and protein array. Flow …
induce type 1 diabetes. Bone marrow (BM)-derived cells were used for colony-formation
assay, quantification of side population (SP) cells, examination of gene expression, nitric
oxide measurement and migration studies. Endothelial progenitor cells (EPCs), a population
of vascular precursors derived from HSCs, were compared in diabetic and control mice.
Cytokines were measured in BM supernatant fractions by ELISA and protein array. Flow …
Methods
Wild-type, gp130−/− and GFP chimeric mice were treated with streptozotocin to induce type 1 diabetes. Bone marrow (BM)-derived cells were used for colony-formation assay, quantification of side population (SP) cells, examination of gene expression, nitric oxide measurement and migration studies. Endothelial progenitor cells (EPCs), a population of vascular precursors derived from HSCs, were compared in diabetic and control mice. Cytokines were measured in BM supernatant fractions by ELISA and protein array. Flow cytometry was performed on enzymatically dissociated retina from gfp+ chimeric mice and used to assess BM cell recruitment to the retina, kidney and blood.
Results
BM cells from the 12-month-diabetic mice showed reduced colony-forming ability, depletion of SP-HSCs with a proportional increase in SP-HSCs residing in hypoxic regions of BM, decreased EPC numbers, and reduced eNos (also known as Nos3) but increased iNos (also known as Nos2) and oxidative stress-related genes. BM supernatant fraction showed increased cytokines, GP130 ligands and monocyte/macrophage stimulating factor. Retina, kidney and peripheral blood showed increased numbers of CD11b+/CD45 hi/CCR2+/Ly6C hi inflammatory monocytes. Diabetic gp130−/− mice were protected from development of diabetes-induced changes in their HSCs.
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