Objective To investigate the effect of M2 microglia (M2-MG) transplantation on spinal cord injury (SCI) repair in mice. Methods Primary MG were obtained from the cerebral cortex of 15 C57BL/6 mice born 2-3 days old by pancreatic enzyme digestion and identified by immunofluorescence staining of Iba1. Then the primary MG were co-cultured with interleukin 4 for 48 hours (experimental group) to induce into M2 phenotype and identified by immunofluorescence staining of Arginase 1 (Arg-1) and Iba1. The normal MG were harvested as control (control group). The dorsal root ganglion (DRG) of 5 C57BL/6 mice born 1 week old were co-cultured with M2-MG for 5 days to observe the axon length, the DRG alone was used as control. Forty-two 6-week-old female C57BL/6 mice were randomly divided into sham group (n=6), SCI group (n=18), and SCI+M2-MG group (n=18). In sham group, only the laminae of T10 level were removed; SCI group and SCI+M2-MG group underwent SCI modeling, and SCI+M2-MG group was simultaneously injected with M2-MG. The survival of mice in each group was observed after operation. At immediate (0), 3, 7, 14, 21, and 28 days after operation, the motor function of mice was evaluated by Basso Mouse Scale (BMS) score, and the gait was evaluated by footprint experiment at 28 days. The spinal cord tissue was taken after operation for immunofluorescence staining, in which glial fibrillary acidic protein (GFAP) staining at 7, 14, and 28 days was used to observe the injured area of the spinal cord, neuronal nuclei antigen staining at 28 days was used to observe the survival of neurons, and GFAP/C3 double staining at 7 and 14 days was used to observe the changes in the number of A1 astrocytes. Results The purity of MG in vitro reached 90%, and the most of the cells were polarized into M2 phenotype identified by Arg-1 immunofluorescence staining. M2-MG promoted the axon growth when co-cultured with DRGs in vitro (P<0.05). All groups of mice survived until the experiment was completed. The hind limb motor function of SCI group and SCI+M2-MG group gradually recovered over time. Among them, the SCI+M2-MG group had significantly higher BMS scores than the SCI group at 21 and 28 days (P<0.05), and the dragging gait significantly improved at 28 days, but it did not reach the level of the sham group. Immunofluorescence staining showed that compared with the SCI group, the SCI+M2-MG group had a smaller injury area at 7, 14, and 28 days, an increase in neuronal survival at 28 days, and a decrease in the number of A1 astrocytes at 7 and 14 days, with significant differences (P<0.05). ConclusionM2-MG transplantation improves the motor function of the hind limbs of SCI mice by promoting neuron survival and axon regeneration. This neuroprotective effect is related to the inhibition of A1 astrocytes polarization.
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 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 morphological and functional changes of retinal degeneration in mice with CLN7 neuronal ceroid-lipofuscinosis, and the therapeutic effects of glial cell derived neurotrophic factor (GDNF) and/or ciliary neurotrophic factor (CNTF) based on neural stem cells (NSC) on mouse photoreceptor cells. MethodsA total of 100 CLN7 mice aged 14 days were randomly divided into the experimental group and the control group, with 80 and 20 mice respectively. Twenty C57BL/6J mice aged 14 days were assigned as wild-type group (WT group). Mice in control group and WT group did not receive any interventions. At 2, 4, and 6 months of age, immunohistochemical staining was conducted to examine alterations in the distribution and quantity of cones, rod-bipolar cells, and cone-bipolar cells within the retinal of mice while electroretinography (ERG) examination was utilized to record scotopic a and b-waves and photopic b-wave amplitudes. At 14 days of age, the mice in the experimental group were intravitreally injected with 2 μl of CNTF-NSC, GDNF-NSC, and a 1:1 cell mixture of CNTF-NSC and GDNF-NSC (GDNF/CNTF-NSC). Those mice were then subdivided into the CNTF-NSC group, the GDNF-NSC group, and the GDNF/CNTF-NSC group accordingly. The contralateral eyes of the mice were injected with 2 μl of control NSC without neurotrophic factor (NTF) as their own control group. At 2 and 4 months of age, the rows of photoreceptor cells in mice was observed by immunohistochemical staining while ERG was performed to record amplitudes. At 4 months of age, the differentiation of grafted NSC and the expression of NTF were observed. Statistical comparisons between the groups were performed using a two-way ANOVA. ResultsCompared with WT group, the density of cones in the peripheral region of the control group at 2, 4 and 6 months of age (F=285.10), rod-bipolar cell density in central and peripheral retina (F=823.20, 346.20), cone-bipolar cell density (F=356.30, 210.60) and the scotopic amplitude of a and b waves (F=1 911.00, 387.10) in central and peripheral retina were significantly decreased, with statistical significance (P＜0.05). At the age of 4 and 6 months, the density of retinal cone cells (F=127.30) and b-wave photopic amplitude (F=51.13) in the control group were significantly decreased, and the difference was statistically significant (P＜0.05). Immunofluorescence microscopy showed that the NSC transplanted in the experimental group preferentially differentiated into astrocytes, and stably expressed CNTF and GDNF at high levels. Comparison of retinal photoreceptor nucleus lines in different treatment subgroups of the experimental group at different ages: CNTF-NSC group, at 2 months of age: the whole, central and peripheral regions were significantly different (F=31.73, 75.06, 75.06; P＜0.05); 4 months of age: The difference between the whole area and the peripheral region was statistically significant (F=12.27, 12.27; P＜0.05). GDNF/CNTF-NSC group, 2 and 4 months of age: the whole (F=27.26, 27.26) and the peripheral area (F=16.01, 13.55) were significantly different (P＜0.05). In GDNF-NSC group, there was no statistical significance at all in the whole, central and peripheral areas at different months of age (F=0.00, 0.01, 0.02; P＞0.05). ConclusionsCLN7 neuronal ceroid-lipofuscinosis mice exhibit progressively increasing degenerative alterations in photoreceptor cells and bipolar cells with age growing, aligning with both morphological and functional observations. Intravitreal administration of stem cell-based CNTF as well as GDNF/CNTF show therapeutic potential in rescuing photoreceptor cells. Nevertheless, the combined application of GDNF/CNTF-NSC do not demonstrate the anticipated synergistic protective effect. GDNF has no therapeutic effect on the retinal morphology and function in CLN7 neuronal ceroid-lipofuscinosis mice.
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.
Objective To observe the effects of the bone marrow mesenchymal stem cells (BMSCs) on the expression of neurotrophic factor protein gene in the retinal detachment (RD) rabbits. Methods 60 healthy rabbits were randomly divided into control group (group A), retinal detachment with PBS group (group B), retinal detachment with BMSCs group (group C), 20 rabbits in each group. RD model were established for rabbits in group B and C. 10 μl PBS was injected into the subretinal space of rabbits in group B, while 10 μl CM-Dil labeled BMSC PBS was injected into subretinal space of rabbits in group C. The rabbits in the group A received no treatment. At 1, 2 and 4 weeks after modeling, the mRNA expression of basic fibroblast growth factor (bFGF), brain derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) were measured by real-time quantitative PCR. Results At 1, 2 and 4 weeks after modeling, the mRNA expression of bFGF, BDNF, CNTF on retinal tissue were increased significantly in group C as compared with group A and B (P < 0.01). At 1 week after modeling, the mRNA expression of bFGF and CNTF on retinal tissue were increased significantly in group B as compared with group A, the mRNA expression of BDNF on retinal tissue in group B was similar with group C. At 2 and 4 weeks after modeling, the mRNA expression of bFGF, BDNF, CNTF were decreased in group B as compared with group A. Conclusion Subretinal transplantation of BMSC can increase the mRNA expression of bFGF, BDNF and CNTF on retinal tissue in RD rabbits.
ObjectivesTo systematically review the efficacy and safety of autologous bone marrow cells therapy for patients with diabetic foot. GRADE system was used to evaluate the evidence quality of outcomes.MethodsWe searched databases including PubMed, EMbase, The Cochrane Library, CBM, WanFang Data and CNKI for randomized controlled trials (RCTs) about bone marrow cell transplantation in patients with diabetic foot from inception to February 28th 2017. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Meta-analysis was performed using RevMan 5.3 software. The evidence quality was evaluated by using GRADE profiler 3.6 software.ResultsFour RCTs were included. Meta-analysis showed that the bone marrow cell transplantation could decrease the rate of amputation (RR=0.08, 95%CI 0.00 to 1.32, P=0.08) and rest pain score (MD=–1.89, 95%CI –2.24 to –1.55, P<0.000 01), increase the rate of ameliorate ulcer healing (RR=2.01, 95%CI 1.45 to 2.79,P<0.000 1) and the quantity of new collateral vessels (MD=1.33, 95%CI 0.60 to 2.05,P<0.000 3). Besides, bone marrow cell transplantation could improve ankle-brachial index (MD=0.16, 95%CI 0.10 to 0.22,P<0.000 01) and transcutaneous arterial oxygen tension (MD=18.81, 95%CI 16.06 to 21.57,P<0.000 01). No adverse event was reported for all included studies. The qualities of evidence for all outcomes were rated as "low" to "very low".ConclusionBased on the current evidence, autologous bone marrow cells transplantation therapy has a certain effect and it is safe for patients with diabetic foot. However, due to the limited quantity and quality of included studies, the above conclusions are still needed more multicenter clinical trials with large sample size to confirm.
Objective To assess systematically the safety and ef fects of stem cell transplantation in stroke patients.Methods CENTRAL (April 2007), MEDLINE (1966 to April 2007), EMBASE (1980 to April 2007), and other databases were searched for RCT of the use of stem cell transplantation for patients with stroke. We critically appraised the quality of included studies according to Juny 2001. We assessed the effects of stem cell therapy on mortal ity, functional outcomes, cognitive functions, image changes, quality of life, and adverse effects by doing meta-analysis with The Cochrane Collaboration’ s Review Manager. Dichotomous outcomes were reported as relative risk and continuous outcome measures as weighted mean differences, with 95% confidence intervals.Results Three RCTs and one historical controlled trial were included involving a total of 69 participants. Only one trial reported the effect on mortality, but because of the small number of death it was not possible to detect any significant differences between stem cell transplantation and routine treatment (RR 0.11, 95%CI 0.01 to 2.31, P = 0.16). Three studies indicated a statistically significant improvement of some functional outcomes in patients treated by stem cell transplantation. Improvements of cognitive function were reported in another trial. One trial showed that the stem cell transplantation significantly improved qual ity of life compared with the control group. Conclusion The current evidence is insufficient to determine whether or not stem cell transplantation is a safe and effective therapy for stroke patients. High-quality, large-scale randomized trials are needed to assess the role of stem cell transplantation for stroke.
Replacement therapy of stem cells transplantation represents a potential treatment for neural retinal diseases. Despite the encouraging results in laboratory, the clinical application of cells replacement therapy is still difficult because the limitation of seed cells, immunologic rejection, oncogenicity and ethical problems, etc. Recent breakthrough in somatic reprogramming provides a promising solution overcoming these obstacles. Further researches on virus free reprogramming will make the clinical application of stem cell replacement therapy possible.