MRI Tracking Study of Transplanted Bone Marrow Stromal Cells Labeled with Resovist in Rat Models with Parkinson’s Disease in Vivo
|Course||Medical Imaging and Nuclear Medicine|
|Keywords||Bone Marrow Stromal Cells Parkinson’s Disease Resovist Magnetic Resonance Imaging tracking Reduced Glutathione|
Parkinson’s disease(PD),an age-associated neurodegenerative disease,is characterized pathologically by degeneration of the dopaminergic neurons of the nigrostriatal pathway and a dramatic loss of dopaminergic neurons(DN) in the substantia nigra(SN) of the ventral midbrain,and the subsequent deficiency of dopamine in the brain areas.It is manifested as motor impairment with characteristic symptoms(extrapyramidal symptoms)including difficulty in initiating movement,resting tremor,rigidity and postural instability.However,until now,very little is known about why and how the PD neurodegenerative process begins and progresses,so the traditional treatments are all symptomatic treatments,including drug treatment and operation,which could improve the symptoms to some extent,but couldn’t prevent the development of the course.In recent years,with the development of some newly rising subjects such as molecular biology and transplantation immunology,many scholars pay attention to a new treatment which is cellular transplantation therapy,and many observations have strongly suggested that it can be a promising therapeutic option for patients with PD.Bone Marrow Stromal Cells (BMSCs) are stem cells with the characteristics of self-renewing,easily isolating and expanding in vitro and its use has no ethical and immunological problems.Furthermore,it can be induced to differentiate into neural cells to substitute impaired DN.Therefore, BMSCs are ideal target cells for cell transplantation.Now there are a few studies about the therapy of BMSCs in PD,and people pay more attention to the methods of tracking transplanted cells in vivo.It seems that the most important task is to determine the optimal transplanted dosage of BMSCs to treat PD and track in vivo.The amount of GSH(reduced glutathione hormone) in the SN of Parkinsonian is decresed and in direct correlation with the pathogenetic condition.Some studies show that injecting GSH intraperitoneally after transplanting BMSCs in the corpus striatum of rat models with PD may has neuroprotective effect.But it is uncert whether GSH could promoting the migration of transplanted BMSCs or not.In our study,we labeled the BMSCs harvested from Resovist-a type of SPIO,and transplanted different quantity of the SPIO-labeled BMSCs into the corpus striatum of rat models of PD and evaluated the distributions of SPIO-positive cells in the brain in different time after transplantation to evaluate the feasibility of visualization of BMSCs with magnetic resonance imaging(MRI) and determine the optimal quantity of transplanted BMSCs with 1.5T MRI and micro-47 coil in vivo.Also,in our study,we evaluate the effect of reduced glutathione and BMSCs in the therapy of rat models of PD.PartⅠMRI Tracking Study of Transplanted Bone Marrow Stromal Cells Labeled with Resovist in Rat Models with Parkinson’s Disease in VivoPurpose:To observe the migration ability of different quantity of BMSCs labled with Resovist after transplanted in rat models with PD with 1.5T MR scanner and micro-47 coil to determine the optimal transplanted dosage of BMSCs and observation time in vivo.Methods:(1) To make rat models with PD;(2) To prepare BMSCs;(3) To lable BMSCs with Resovist and Brdu in vitro;(4) To divide into groups:40 PD rats were randomly assigned to 5 groups,ie,control group,1×10~5 BMSCs group,1.5×10~5 BMSCs group,2×10~5 BMSCs group and 2.5×10~5 BMSCs group,with 8 rats in each group.(5) To transplant BMSCs labled with Resovist and Brdu in the corpus striatum of rat models with PD;(6) To trace with MRI in vivo:TSE-T1WI、TSE-T2WI and FFE-T2WI were obtained immediately and 1week、4 weeks、8 weeks after transplantation with 1.5 Tesla 47mm inner diameter micro-coil.Selecting the most sensitive sequence to measure and compare the volume and signal intensity of hypointense areas of different groups in different time. (7) Apomorphine rotation test:rotational behavior was assessed in each group immediately and 1week、4 weeks、8 weeks after transplantation.(8) Pathology examination:after MRI the rats were executed,the brain tissue around corpus striatum were analyzed histologically and prepared for immunohistochemistry staining and real time quantitative reverse transcription PCR at 12 weeks after transplantation.Results:(1) FFE-T2WI sequence is the most sensitive sequence to evaluate the migration of BMSCs with 1.5T Tesla 47mm inner diameter micro-coil in vivo.(2)The injected sites containing labeled cells could all be detected as well-defined hypointense areas(dark region) through MRI and were confirmed on pathology.(3)Only the extent of the dark region becomes wider at 4 weeks after transplantation in 2×10~5 BMSCs group. (4)The therapeutic efficacy of 2×10~5 BMSCs group was the best which was confirmed on behaviour studies and real time quantitative reverse transcription PCR.Conclusion:(1) It is feasible to magnetically label and visualize BMSCs in vivo. FFE-T2WI sequence is the most sensitive sequence.(2) For the treatment of PD and evaluation of the migratory ability of BMSCs,the optimal transplanted dosage of BMSCs was 2×10~5 while the best observation time was 4 weeks after transplantationPartⅡMRI Tracking Study of Transplanted Bone Marrow Stromal Cells Labeled with Resovist and Injected GSH Intraperitoneally in Rat Models with Parkinson’s Disease in VivoPurpose:To observe the neuroprotective effect of reduced glutathione in a rat Parkinson’s disease model treated with transplanted BMSCs with 1.5T MR and micro-47 coil.Methods:(1) To transplant 2.0×10~5 BMSCs into right corpus striatum of PD rats at the base of partⅠ.(2)18 PD rats were randomly assigned to 3 groups,ie,control group(n= 4),BMSCs+reduced glutathione group(to inject reduced glutathione intraperitoneally at 2 weeks after transplantation of BMSCs,once a day,n=7) and BMSCs group(n=7).(3) FFE-T2WI were obtained immediately and 1week、2 weeks、4 weeks after the intraperitoneal injection of reduced glutathione with 1.5 Tesla 47mm inner diameter micro-coil.At the meantime,rotational behavior was assessed in each group.(4)After MRI the rats were executed,the brain tissue around corpus striatum were analyzed histologically and prepared for immunohistochemistry staining and real time quantitative reverse transcription PCR.Results:(1) The injected sites containing labeled cells could all be detected as well-defined hypointense areas(dark region) through MRI.(2)The extent of the dark areas of BMSCs+reduced glutathione group is a little wider than that of BMSCs group at 2 and 4 weeks after the intraperitoneal injection of reduced glutathione,but there is no significant difference between them.(3)The therapeutic efficacy of BMSCs+reduced glutathione group was the best which was confirmed on behaviour studies and real time quantitative reverse transcription PCR.Conclusion:(1) The neuroprotective effect of reduced glutathione could improve the behaviour symptoms of PD rats through inducing BMSCs differenciate into neural cells. (2)The reduce glutathione(125mg/kg) maybe couldn’t promote the migration of transplanted BMSCs or the effect is weak,1.5T MRI and immunohistochemistry staining displayed no significant difference between BMSCs+reduced glutathione group and BMSCs.