Objective To observe the retinal apoptosis of laser-induced retinal injury in mice after bone marrow mesenchymal stem cells transplantation. Methods Green fluorescent protein (GFP) labeled MSCs from C57BL/6 mice were cultured in vitro. A total of 135 C57BL/6 mice were divided into three groups including normal control group (15 mice), injured control group (60 mice) and MSCs treatment group (60 mice). Laser retinal injuries were induced by laser photocoagulation. One day after photocoagulation, 02 ml cell suspension, which contained 1times;106 GFP-MSCs, were injected into the mice in treatment group via tail vein, and the mice in injured control group were given equal volume of phosphate buffer solution. Animal were execute on three, seven, 14 and 21 days following laser damage. Hematoxylin and eosin (HE) staining was performed to assess the changes of injured retinas. The diameters of laser spots and areas with total loss of cells in outer nuclear layer (ONL) were analyzed by image processing software. The apoptosis of retinal cells was examined by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) staining. The migration of GFP-MSCs into the retina was observed by fluorescence microscope. Results HE staining showed that the retinal structures were integrated in normal control group. Retinal damages were observed both in injured control group and MSCs treatment group, but milder in the latter. Though the average diameter of area with total loss of cells in ONL of MSCs treatment group was less than the injured control group (t=5.769, P＜0.05), the diameters of laser spots show no difference (t=0.964,P＞0.05) on day three. Both the average diameter of laser spots (t=5.180, 5.417, 2.381) and area with total loss of cells in ONL (t=3.530, 3.224, 3.162) were less in the MSCs treatment group on day seven, 14 and 21 (P＜0.05). TUNEL staining shows that the apoptosis were decreased after MSCs transplantation on day three, seven, 14 and 21 (t=11.142, 7.479, 6.678, 3.953,P＜0.05). No apoptosis was observed in normal control group. Very few GFP-MSCs were observed in the retina at all time-points. They were only seen in the subretinal and choroidal neovascularization occasionally on day seven and 14. Conclusion MSCs transplantation can effectively limit the range of retinal laser damage and inhibit cell apoptosis.
Objective To observe the effects of subretinal transplantation of rat mesenchymal stem cells (rMSCs) on Sodium Iodate (SI)induced retinal degeneration. Methods One hundred and twenty BrownNorway (BN) rats were divided into three groups including SI injection group,rMSCs transplantation group and normal control group, each with 40 rats. The retinal degeneration was induced by caudal vein injection of SI. The retinal pigment epithelium（RPE）and neural retinal were evaluated by ocular fundus photograph, fluorescein fundus angiography (FFA)，electroretinogram (ERG) and histological approach, and TUNEL（terminal deoxynucleotidyl transferasemediated dUTP nick end labeling ）. CMDiIprelabeled primary rMSCs were transplanted into the subretinal space of SIinduced rats. The survival, integration, and differentiation of rMSCs were observed between 14 day to 60 day after the transplantation.Results The rat retinal function was gradually reduced after14 days of SI injection, with a timedependent manner. After the RPE cells were damaged，the outer segments of photoreceptors became disrupted and shortened until karyopyknosis. The nuclear morphology and positive TUNEL labeling indicated that the death of photoreceptor cells was apoptosis. After rMSCs transplantation, CMDiI labeled donor cells were observed to be scattered in the subretinal space and expressed RPE cell markers. Average amplitude of b wave and Ops (oscillation potential) in ERG improved 27.80%,59.38% respectively after rMSCs transplantation.Conclusions Transplanted rMSCs can survive in subretinal space and differentiate into RPE.
Objective To observe the survival of human umbilical cord derived mesenchymal stem cells (hUC-MSCs) after injection into the vitreous of rabbits,and the animal safety under those procedures.Methods Twentyseven pigmented rabbits were randomly divided into 3 groups (intravitreal injection 1 week group，2 weeks group and 4 weeks group)， each with 9 rabbits.For each animal the right eye was the experimental eye receiving hUCMSCs injection,while the left eye was the control eye receiving culture medium. The rabbit eyes were examined by slitlamp microscope, indirect ophthalmoscopy, fundus photography, fundus fluorescence angiography（FFA）and Tonopen tonometer before and after injection. hUCMSCs were labeled by CMDil in vitro, and their survival status was measured by confocal fluorescence microscopy, light microscope and transmission electron microscope at 4 weeks after injection. Results Four weeks after injection, a large number of the hUCMSCs were still alive in the vitreous cavity. The overall condition of those rabbits was good. The anterior segment and retina of experimental eyes were normal, without hyperfluorescence, hypofluorescence and leakage in the retina at 1,2 and 4 weeks after injection. There was no significant difference on IOP before and after injection at different time points (P＞0.05), and no obvious changes at cornea, anterior chamber angle,lens,retinal structure by.light microscope and transmission electron microscope examination.Conclusion hUC-MSCs can survive in the rabbit vitreous for four weeks；intravitreal injection of hUCMSCs was safe and feasible.
Objective To assess the effectiveness and safety of autologous stem cell transplantation after high-dose chemotherapy in first-line treatment of follicular lymphoma. Method Randomized controlled trials (RCTs) of autologous stem cell transplantation after high-dose chemotherapy in first-line treatment of follicular lymphoma were collected from MEDLINE (1990-2009), EMBASE (1990-2009), OVID (1990-2009), and the Cochrane Library (Issue 2, 2009), and the proceedings of ASH were searched manually. The methodological quality of included studies was evaluated, and data analysis was performed with software STATA 10.0 and RevMan 4.3. Result A total of 4 RCTs involving 941 patients were included. The results of meta-analysis showed that overall survival rate (HR=0.82, 95%CI 0.49 to 1.15), event-free survival rate (HR=0.35, 95%CI 0.24 to 0.47), total remission rate (RR=0.35, 95%CI 0.96 to 1.30), and secondary malignant tumor incidence rate (RR=1.68, 95%CI 0.47 to 6.07). Conclusion According to the present evidences, autologous stem cell transplantation after high-dose chemotherapy can not improve overall survival rate and total remission rate, but can improve event-free survival rate, and do not increase secondary malignant tumor incidence rate. However, more high-quality, multiple-center, large-sample randomized controlled trials are required.
ObjectiveTo investigate the behavioral recovery of spinal cord injury (SCI) rats that received transplantation of NEP1-40 gene-modified neural stem cells. MethodsNeural stem cells (NSCs) were derived from the cortex tissue of rat embryo at the age of 18 days and identified by Nestin immunofluorescence. The lentiviruses were transduced to NSCs to construct NEP1-40 gene modified NSCs. Spinal cords of 30 Sprague-Dawley rats were hemisected at the nineth thoracic vertebrae level. The rats were randomly assigned to three groups. Cell culture medium, NSCs and NEP1-40 gene-modified NSCs were transplanted into the lesion site of rats of SCI group, NSCs group and NEP1-40-NSCs group respectively 7 days after injury. Additional 10 rats served as blank control group (sham group), which only received laminectomy. Following transplantation, behavior tests including Basso, Beattie, Bresnahan (BBB) Locomotor Rating Scale and grid test were utilized to evaluate spinal cord functional recovery. ResultsBehavior tests 8 weeks after cells transplantation showed that the rats in SCI group got worst results, the BBB scores improved and the grid drop times reduced significantly in NSCs transplantation group (P<0.01) and behavioral test outcomes were best in the NEP1-40 gene-modified NSCs group (P<0.01). ConclusionNEP1-40 gene modification can significantly improve the behavioral recovery of SCI rats that received transplantation of pure neural stem cells. It can provide a new idea and reliable experimental base for the study of NSCs transplantation for spinal cord injury.
ObjectiveTo systematically review the efficacy and safety of non-myeloablative stem cell transplantation (NST) for the treatment of multiple myeloma (MM) after first autologous stem cell transplantation. MethodsSuch databases as The Cochrane Library (Issue 5, 2013), PubMed, EMbase, CBM, CNKI, VIP and WanFang Data were electronically searched to collect studies investigated the efficacy and safety of NST and non-NST for the treatment of MM after first autologous stem cell transplantation from the date of their establishment to June 13th 2013. Two reviewers independently screened studies according to the inclusion and exclusion criteria, extracted data and evaluated the methodological quality of the included studies. Then meta-analysis was performed using RevMan 5.1 software. ResultsSeven studies involving 1 961 participants were included, of which 626 cases were in the NST group and 1 335 cases were in the non-NST group. The results of meta-analysis showed that no significant difference was found between both groups in the overall survival rate (HR=1.06, 95%CI 0.78 to 1.44, P=0.69) and progress-free survival rate (HR=0.92, 95%CI 0.76 to 1.11, P=0.39). However, there were significant differences in the complete remission rate (RR=1.29, 95%CI 1.13 to 1.48, P=0.000 2) and treatment-related mortality rate (RR=3.40, 95%CI 2.27 to 5.07, P < 0.000 01). ConclusionThe efficacy of NST is not superior to non-NST for patients with MM which has received first autologous stem cell transplantation. It is not sufficient to recommend NST as the first-line treatment of MM based on the currently available evidence.
ObjectiveTo systematically review clinical efficacy and safety of bone marrow stem cells transplantation in treating primary dilated cardiomyopathy (DCM). MethodsSuch databases as PubMed, CENTRAL, EMbase, Web of Knowledge, VIP, CNKI, CBM and WanFang Data were searched from inception to March 2014 for the randomized controlled trials (RCTs) about bone marrow stem cells transplantation for DCM. According to the inclusion and exclusion criteria, two reviewers independently screened literature, extracted data, and assessed methodological quality of included studies. Then meta-analysis was performed using RevMan 5.2.0 software. ResultsA total of ten RCTs involving 374 patients were included. The results of meta-analysis showed that, a) for safety, after 3 months there was no significant difference in the incidence of malignant arrhythmia events between bone marrow stem cell transplantation group and routine treatment group (RR=0.81, 95%CI 0.38 to 1.72, P=0.58); and b) for efficacy, compared with the control group, left ventricular ejection fraction (LVEF) increased in the bone marrow stem cell transplantation group after 3 months (WMD=3.86, 95% CI 2.53 to 5.20, P<0.000 01) and after 6 months (WMD=5.54, 95%CI 3.02 to 8.06, P<0.000 1). The bone marrow stem cell transplantation group were better in increased 6-minute walking distance after 3 months (WMD=22.12, 95%CI 7.78 to 36.46, P=0.003), increased 6-minute walking distance after 6 months (WMD=102.79, 95%CI 50.16 to 155.41, P=0.000 1), decreased perfusion defect of myocardium percentage after 3 months (WMD=-4.00, 95%CI -5.87 to -2.13, P<0.000 1). However, there was no significant difference in left ventricular end-diastolic diameter (LVEDD) between two groups after 3 months (WMD=-0.37, 95%CI -1.67 to 0.93, P=0.57) and after 6 months (WMD=-0.70, 95%CI -2.76 to 1.36, P=0.51). ConclusionBone marrow stem cells transplantation for dilated cardiomyopathy is effective in improve patients' heart function with good safety, with significant difference. Due to limited quantity and quality of the included studies, more high quality and large-scale RCTs are needed to verify the above conclusion.
ObjectiveTo observe the effects of human umbilical cord mesenchymal stem cells (hUCMSCs) on blood glucose levels and diabetic retinopathy in diabetes mellitus (DM) rats. MethodA total of 45 healthy male Sprague-Dawley rats were randomly divided into normal control group (group A, 10 rats) and DM group (33 rats). Diabetic model was established in DM group by tail vein injection of streptozotocin.The DM group was further randomly divided into 3 groups (11 rats in each group), including group B (no transplantation), group C (hUCMSC was injected through tail vein) and group D (hUCMSC was injected into the vitreous). Blood glucose, retina wholemont staining and expression of brain derived neurotrophic factor (BDNF) in the retina were measured at 2, 4, 6, 8 weeks after hUCMSC injection. The blood glucose was significantly different between A-D groups before injection (t=-64.400, -60.601, -44.065, -43.872; P=0.000) BDNF expression was studied by real time fluorescence quantitative polymerase chain reaction (RT-PCR) and immunohistochemistry staining. ResultsThe blood glucose was significantly different between A-D groups after hUCMSC injection (F=400.017, 404.410, 422.043, 344.109; P=0.000), and between group C and group B/D (t=4.447, 4.990; P < 0.01). Immuno-staining shown that BDNF was positive in ganglion cell layer (RGC) of group A, weak in group B while BDNF expression increased in group C/D. BDNF mRNA expression was significantly different between group B, C and D at 4, 6 and 8 weeks after hUCMSC injection (F=29.372, 188.492, 421.537; P=0.000), and between group B and C/D (t=66.781, 72.401, 63.880, 88.423, 75.120, 83.002; P < 0.01) by RT-PCR analysis. The BDNF mRNA expression was significantly different between C and D groups only at 8 weeks after hUCMSC injection (t=127.321, P=0.005). ConclusionsTail vein injection of hUCMSCs can significantly reduce the blood glucose levels of rats. Intravenous and intravitreal injection of hUCMSCs can increase the expression of BDNF.
ObjectiveTo observe the protective effect of human umbilical cord blood stem cells (hUCBSC) transplantation on retinal ganglion cells (RGC) after optic nerve injury. Method48 adult Sprague-Dawley rats were randomly divided into group A and B, therefore 24 rats in each group. Calibrated optic nerve crush injury model was induced in the left eyes, the right eyes served as a control. Medicine was injected at seventh day after optic nerve injury. PBS was injected into the eyes of Group A rats by peribulbar injection. The hUCBSCs were injected into the eyes of Group B rats by peribulbar injection. Seven days before sacrifice, 5% fluorogold was injected into superior colliculi bilaterally. At 7, 14, 21, 28 days after labeled, retinal flat mounts were observed under fluorescence microscope and optical microscope to investigate the morphological and RGC changes in density during retinal degeneration. ResultsThe RGC number showed a tendency to decline gradually along with increases of the time in two groups, but the trend of decrease of Group B was evidently slower than that of Group A. The RGC number of the injury eye were less than the control eye in Group A and B (t=3.24, 3.15; P < 0.05). At 7, 14, 21, 28 days after labeled, the RGC number (t=4.78, 4.70, 3.98, 3.27; P < 0.05) and labeled RGC rate (t=4.39, 4.21, 4.36, 5.07; P < 0.05) in group B were more than those in group A. After optic nerve injury, there was karyopycnosis on ganglion cell layer of retina, thinning on each layer of retina, derangement of cell and decrease in RGC. There was different degree of the above change in different time after optic nerve injury. There were the swelling, the hemorrhage, derangement of spongiocyte and the denaturation like vacuole in the spot of optic nerve injury. Moreover, they were aggravating with increases of the time after optic nerve injury. There was no pathological changes in normal eyes. ConclusionThe hUCBSC can increase the survival rate of the RGC and can rescue and(or) restore the injujed RGC after transplanted into body of optic nerve crush rat model by peribulbar injection.