Chemotactic Activities of Human Bone Marrow Mesenchymal Stem Cells Was Inhibited Following Incubationwith Diabetes Mellitus Type II Serum

Background: Type ۲ diabetes mellitus (DM۲) are susceptibleto progress associated cardiovascular disease, abnormal angiogenesis,and vascular bed complications. It was previously revealedthat specific bone marrow stem/progenitor populationswere more sensitive to a persistent diabetic condition whichassociated with numerous fundamental defects. Accumulatingdata support the hypothesis that bone marrow mesenchymalstem cells (BM-MSCs) could migrate and home to the woundarea to promote angiogenesis by various mechanisms. We investigatedthe effect of DM۲ sera on chemotactic activities ofhuman bone marrow mesenchymal stem cells in vitroMaterials and Methods: BM-MSCs were allocated into threegroups; control (DMEM with ۱۰% FBS); non-diabetic (DMEMwith ۱۰% normal human serum), and diabetic (DMEM with۱۰% diabetic serum) groups. After ۷ days treatment, we performedan in vitro migration assay by using ۸-μm pore sizeTranswell membrane inserts. BM- MSCs were resuspended in۲۰۰ μl DMEM containing ۲% diabetic or non- diabetic serumand transferred into the Transwell insert. In control group, cellswere only exposed to ۲% FBS. In the lower basolateral chamber,۷۰۰ μl of DMEM enriched with ۵۰ nM SDF۱-α or ۱۰ ng/ml VEGF was added. After incubation at ۳۷˚C for ۲۴ h, thenumber of migrated cells was counted in ۵ random high-powerfields. In order to verify the results of migration/chemotacticassay, the expression of CXCR-۴, VEGFR-۲, and VEGF werequantified by real- time PCR system with the SYBR Green PCRMaster Mix. Data were analyzed using one way ANOVA andTukey's test and the means were considered significantly differentat P<۰.۰۵.Results: In factor free condition, the number of migrated BMMSCsafter exposure to diabetic condition was significantlydiminished as compared with non-diabetic and control groups(pdiabetic sera versus control<۰.۰۰۰۱; pdiabetic versus nondiabeticsera<۰.۰۵). Similar to factor-free condition, a markedreduction was observed in diabetic BM-MSCs in responseto SDF-۱α (pdiabetic versus control <۰.۰۱; pdiabetic versusnon-diabetic groups<۰.۰۵). Compared with either control ornon-diabetic groups, diabetic sera abolished the migration ofBM-MSCs in response to VEGF (pdiabetic sera versus controland non-diabetic groups<۰.۰۰۰۱). Compared to controlgroup, the mRNA level of CXCR-۴ gene in non-diabetic anddiabetic group was significantly reduced (pcontrol versus diabeticand non-diabetic sera<۰.۰۰۱). Similar to expression patternof CXCR- ۴, DM۲ had potential to reduce the expression ofVEGFR-۲ (pdiabetic sera versus control<۰.۰۵; pdiabetic seraversus non-diabetic sera<۰.۰۱) and VEGF (pdiabetic sera versuscontrol<۰.۰۰۱; pdiabetic versus non-diabetic sera<۰.۰۱) ascompared with control and non- diabetic groups.Conclusion: Our data revealed that DM۲ condition diminishedchemotactic activity of BM- MSCs toward VEGF and SDF-۱α and also DM۲ sera could suppress SDF-۱α/ CXCR-۴ andVEGF/VEGFR-۲ axis in BM-MSCs.

کلیدواژه ها:

Diabetes Type II ، Mesenchymal Stem Cells ، Chemotactic ، Migration