Quantitative Cell Numbers and Density of Mesangial Volume in a Rat Model with Induced Hyperglycemic and Treated with Mononuclear Derived CD133 Positive Cells

There is evidence that mesangial cell structural changes contribute to the pathogenesis of diabetic nephropathy. To gain better insight into the mechanisms responsible for this issue, present study focused on effect of cord blood mononuclear cells (MNCs) derived CD133 positive cells on mesangial cell structure and function. The animals were randomly divided into four groups (each with six rats) and were kept in separate cages as follows: Group I: control group, received only 8.2 mmol/L sodium citrate buffer (pH 5.4). Group II: received only CD133 positive cells. Group III: received alloxan (65mg/kg) only. Group IV: received alloxan, followed by administration of CD133 positive cells, 1 week later. Rats were studied for 16 weeks. Cord blood mononuclear cells (MNCs) were isolated by a conventional centrifuge method through a Ficoll-density gradient, CD133 positive isolation was performed by means of magnetic cell separation (MACS) columns according to the manufacturer’s procedure. CD133 positive stem cells analyzed using flow cytometry. The CD133positive cells were centrifuged, re-suspended with PBS, and transplanted to the rats through the tail. At the end of the experiments, blood was collected, and then blood glucose, creatinine, glycated hemoglobin and insulin concentrations were measured by using kits. All of the animals were killed and the kidneys were removed. Tissues were processed for light microscopy. Glomerular features were evaluated quantitatively using Cavalieri and disectory methods and compared with sham and control groups. Our results indicated that treated hyperglycemic rats showed an increase in mesangial volume compared to untreated group. Concerning the mechanisms of these findings both glycemic control and CD133 positive cells regenerative potential are major’s factors to change mesangial structure and function. The present study clearly documents the potential of CD133 positive cells on the renal mesangial cells.