3.0T MRI Tracing BMSCs Transplantation For The Repair Of Renal Ischemia Reperfusion Injury In Vivo

1. Purposes1.1To explore the morphology and growth characteristics of BMSCs, and to identify whether they are the BMSCs or not.1.2Establish the kidney ischemia-reperfusion injury model, and transplant labled cells(two methods:through renal artery and subcapsular); Tracing cells by MR in vivo, T2and T2*map image can analyse quantitatively.1.3Make comprehensive evaluation to the treatment of BMSCs transplantation.2. Materials and methods2.1Research object40purebred SD rats, male,5to6w,170-200g (provided by Shanxi Medical University Laboratory Animal Center).2.2Experimental MethodsMRI method:Use Siemens Verio3.0T magnetic resonance imaging, supine, head first. Scan sequences:T2WI, T2*WI, multiple-echo T2and T2*sequences. Do scans before the injury and in1d,1w,2w,3w.Cell identification:Use flow cytometry instrument detection, CellQuestPlot software analyse data; Immunofluorescence method:routine immunofluorescence staining, with laser confocal microscope to observe.After3w, SD rats were measured weight, sampled blood by the inferior vena cava and materials.Finally we test renal function and observe biopsy.3. Result3.1Studies in vitroUse bone marrow adherent method to separate BMSC successfully. Identify cell phenotype by immunofluorescence and flow cytometry analysis:CD34(-), CD45(-), CD44(+), CD90(+), CD73(+); BMSCs can be successfully induced and differentiated to osteogenesis and fat cells.3.2MRI tracing in vivo(1) T2WI images show double kidney are symmetric in size and signal, and after IRI, the volume and signal of the left kidney increased.(2) The T2*WI trace cells in vivo:in Id, two different methods shows low signal area (iron) are related to local injection, over time, the artery group displays the left kidney appears low signal area, but in the subcapsular group, part of low signal area is still related to local.(3) Compared with T2value between the injury and two treatment groups:After injury, three groups of T2values are rising, the peak is located in the1d. Over time, the T2values have a downward trend, but the two treatment groups were decreased fleetly, and at the same time, T2values were lower than the injury group, but no obvious difference between the two treatment groups. Compared with T2*value between the three sets:T2*value of the kidney ischemia-reperfusion injury group was stable, which proved to be it has little impact on T2*value. The lowest point of T2*values was located in the2w. At the same time, T2*value of the two treatment groups were lower than the damage, but the trend is not consistent between two groups.3.3Effect evaluation(1) Compared with the weight growth rate, from high to low in turn:the control group, the damage, the treatment;(2)Compared with the renal function, the urea nitrogen and creatinine increased from high to low:the injury group, the treatment and the control, the renal artery group is lower than the subcapsular.(3) Compared with HE staining results:control group kidney tissue structure is normal, the damage group shows a large number of cells swelling, the damage degree of the treatment group is lighter than the damage group.4. Conclusion4.1BMSCs were isolated successfully by the adherent method of whole bone marrow.4.2BMSCs labeled SPIO can be targeted to migrate to kidney ischemia-reperfusion injury.4.3MRI T2WI and T2map images can show the changes of RIRI. T2*WI and T2*map image can trace BMSCs labeled SPIO, But no obvious between the two methods of transplantation.4.4Comprehensive Analysis from the weight growth rate, renal function and HE staining, stem cell has therapeutic effect on the RIRI.