Tracking of iron-labeled human neural stem cells by magnetic resonance imaging in cell replacement therapy for Parkinson’s disease

Human neural stem cells (hNSCs) derived from the ventral mesencephalon are powerful research tools and candidates for cell therapies in Parkinson’s disease. However, their clinical translation has not been fully realized due, in part, to the limited ability to track stem cell regional localization and survival over long periods of time afterin vivo transplantation. Magnetic resonance imaging provides an excellent non-invasive method to study the fate of transplanted cellsin vivo. For magnetic resonance imaging cell tracking, cells need to be labeled with a contrast agent, such as magnetic nanoparticles, at a concentration high enough to be easily detected by magnetic resonance imaging. Grafting of human neural stem cells labeled with magnetic nanoparticles allows cell tracking by magnetic resonance imaging without impairment of cell survival, prolif-eration, self-renewal, and multipotency. However, the results reviewed here suggest that in long term grafting, activated microglia and macrophages could contribute to magnetic resonance imaging signal by engulifng dead labeled cells or iron nanoparticles dispersed freely in the brain parenchyma over time.     

The role of synphilin-1 in the pathogenesis of Parkinson’s disease

1 Introduction Parkinson’s disease (PD) is the second most common neurological disorder to affect approximate 0.2% of overall population and 2% of those over the age of 65. The disease is characterized by a triad of cardinal symptoms, including bradykinesia (slowed movement), resting tremor, and rigidity. Progressive degeneration of the dopaminergic (DAergic) neurons which are mostly located in the sub- stantia nigra pars compacta (SNpc), and the formation of eosinophilic inclusions known a…     

Panax notoginseng saponins influence on transplantation of neural stem cell-derived dopaminergic neurons in a rat model of Parkinson’s disease

BACKGROUND:Dopaminergic neurons differentiated from neural stem cells have been successfully used in the treatment of rat models of Parkinson's disease;however,the survival rate of transplanted cells has been low.Most cells die by apoptosis as a result of overloaded intracellular calcium and the formation of oxygen free radicals.OBJECTIVE:To observe whether survival of transplanted cells,transplantation efficacy.and dopaminergic differentiation from neural stem cells is altered by Panax notoginseng saponins(PNS) in a rat model of Parkinson's disease.DESIGN,TIME AND SETTING:Cellular and molecular biology experiments with randomized group design.The experiment was performed at the Animal Experimental Center,First Hospital of Sun Yat-sen University from April to October 2007.MATERIALS:Thirty-two adult,healthy,male Sprague Dawley rats,and four healthy Sprague Dawley rat embryos at gestational days 14-15 were selected.The right ventral mesenceDhalon was injected with 6-hydroxydopamine to establish a model of Parkinson's disease.6-hydroxydopamine and apomorphine were purchased from Sigma.USA.METHODS:Neural stem cells derived from the mesencephalon of embryonic rats were cultivated and passaged in serum-free culture medium.Lesioned animals were randomly divided into four groups(n=8):dopaminergic neuron,dopaminergic neuron PNS,PNS,and control.The dopaminergic neuron group was iniected with 3 μ L cell suspension containing dopaminergic neurons difierentiated from neural stem cells.The dopaminergic neurons PNS group received 3 μ L dopaminergic cell suspension combined with PNS (250 mg/L).The PNS group received 3 μL PNS(250 mg/L),and the control group received 3 μL DMEM/F12 culture medium.MAIN OUTCOME MEASURES:The rats were transcardially perfused with 4% paraformaldehyde at 60 days post-grafting for immunohistochemistry.The rats were intraperitoneally injected with apomorphine (0.5 mg/kg)to induce rotational behavior.RESULTS:Cell counts of tyrosine hydroxylase-positive neurons in the dopaminergic neuron PNS group were(732±82.6)cells/400-fold field.This was significantly greater than the dopaminergic neuron group [(326±34.8)cells/400-fold field,P<0.01].Compared to the control group,the rotational asymmetry of rats that received dopaminergic neuron transplants was significantly decreased,beginning at 20 days after operation(P<0.0 1).Rotational asymmetry was fugher reduced between 10～60 days post-surgery in the dopaminergic neuron PNS group,compared to the dopaminergic neuron group(P<0.01).CONCLUSION:Panax notoginseng saponins can increase survival and effectiveness of dopaminergic neurons differentiated from neural stem cells for transplantation in a rat model of Parkinson's disease.     

Dysfunction of two lysosome degradation pathways of α-synuclein in Parkinson’s disease: potential therapeutic targets?

Parkinson’s disease (PD) is pathologically characterized by the presence of α-synuclein (α-syn)-positive intra-cytoplasmic inclusions named Lewy bodies in the dopaminergic neurons of the substantia nigra. A series of morbid consequences are caused by pathologically high amounts or mutant forms of α-syn, such as defects of membrane trafficking and lipid metabolism. In this review, we consider evidence that both point mutation and overexpression of α-syn result in aberrant degradation in neurons and microglia, and this is associated with the autophagy-lysosome pathway and endosome-lysosome system, leading directly to pathological intracellular aggregation, abnormal externalization and re-internalization cycling (and, in turn, internalization and re-externalization), and exocytosis. Based on these pathological changes, an increasing number of researchers have focused on these new therapeutic targets, aiming at alleviating the pathological accumulation of α-syn and re-establishing normal degradation.     

Effects of cotransplantated Schwann cells and neural stem cells in a rat model of Alzheimer’s disease

Schwann cells (SCs) are significantly better at promoting neural stem cell (NSCs) proliferation, differentiation and synaptic formation when cocultured with NSCs in vitro, compared with cultured in a single nerve growth factor. The present study transplanted NSCs and SCs into the brain of a rat model of Alzheimer’s disease to investigate the effect of cotransplantation. Results show transplantation of both NSCs alone and NSCs + SCs significantly promoted learning and memory functions in Alzheimer’s disease rats, decreased glial fibrillary acidic protein and calcium binding protein S100β expression, but increased expression of the cholinergic neuron marker choline acetyl transferase mRNA. The effect of NSCs + SCs cotransplantation was, however, more significant. NSCs and SCs cotransplantation significantly reduced the number of astrocytes and increased cholinergic neurons, facilitating the recovery of learning and memory function, compared with NSCs transplantation alone.     

Effect of stromal cell-derived factor-1/CXCR4 axis in neural stem cell transplantation for Parkinson’s disease

Previous studies have shown that neural stem cell transplantation has the potential to treat Parkinson’s disease,but its specific mechanism of action is still unclear.Stromal cell-derived factor-1 and its receptor,chemokine receptor 4(CXCR4),are important regulators of cell migration.We speculated that the CXCR4/stromal cell-derived factor 1 axis may be involved in the therapeutic effect of neural stem cell transplantation in the treatment of Parkinson’s disease.A Parkinson’s disease rat model was injected with 6-hydroxydopamine via the right ascending nigrostriatal dopaminergic pathway,and then treated with 5μL of neural stem cell suspension(1.5×104/L)in the right substantia nigra.Rats were intraperitoneally injected once daily for 3 days with 1.25 mL/kg of the CXCR4 antagonist AMD3100 to observe changes after neural stem cell transplantation.Parkinson-like behavior in rats was detected using apomorphine-induced rotation.Immunofluorescence staining was used to determine the immunoreactivity of tyrosine hydroxylase,CXCR4,and stromal cell-derived factor-1 in the brain.Using quantitative real-time polymerase chain reaction,the mRNA expression of stromal cell-derived factor-1 and CXCR4 in the right substantia nigra were measured.In addition,western blot assays were performed to analyze the protein expression of stromal cell-derived factor-1 and CXCR4.Our results demonstrated that neural stem cell transplantation noticeably reduced apomorphine-induced rotation,increased the mRNA and protein expression of stromal cell-derived factor-1 and CXCR4 in the right substantia nigra,and enhanced the immunoreactivity of tyrosine hydroxylase,CXCR4,and stromal cell-derived factor-1 in the brain.Injection of AMD3100 inhibited the aforementioned effects.These findings suggest that the stromal cell-derived factor-1/CXCR4 axis may play a significant role in the therapeutic effect of neural stem cell transplantation in a rat model of Parkinson’s disease.This study was approved by the Animal Care and Use Committee of Kunming Medical University,China(approval No.SYXKK2015-0002)on April 1,2014.     

Are human dental papilla-derived stem cell and human brain-derived neural stem cell transplantations suitable for treatment of Parkinson's disease?

Transplantation of neural stem cells has been reported as a possible approach for replacing impaired dopaminergic neurons. In this study, we tested the efficacy of early-stage human dental papilla-derived stem cells and human brain-derived neural stem cells in rat models of 6-hydroxydopamine-induced Parkinson’s disease. Rats received a unilateral injection of 6-hydroxydopamine into right medial forebrain bundle, followed 3 weeks later by injections of PBS, early-stage human dental papilla-derived stem cells, or human brain-derived neural stem cells into the ipsilateral striatum. All of the rats in the human dental papilla-derived stem cell group died from tumor formation at around 2 weeks following cell transplantation. Postmortem examinations revealed homogeneous malignant tumors in the striatum of the human dental papilla-derived stem cell group. Stepping tests revealed that human brain-derived neural stem cell transplantation did not improve motor dysfunction. In apomorphine-induced rotation tests, neither the human brain-derived neural stem cell group nor the control groups (PBS injection) demonstrated significant changes. Glucose metabolism in the lesioned side of striatum was reduced by human brain-derived neural stem cell transplantation. [18 F]-FP-CIT PET scans in the striatum did not demonstrate a significant increase in the human brain-derived neural stem cell group. Tyrosine hydroxylase (dopaminergic neuronal marker) staining and G protein-activated inward rectifier potassium channel 2 (A9 dopaminergic neuronal marker) were positive in the lesioned side of striatum in the human brain-derived neural stem cell group. The use of early-stage human dental papilla-derived stem cells confirmed its tendency to form tumors. Human brain-derived neural stem cells could be partially differentiated into dopaminergic neurons, but they did not secrete dopamine.     

Intracerebroventricular transplanted bone marrow stem cells survive and migrate into the brain of rats with Parkinson’s disease

In this study,6-hydroxydopamine was stereotaxically injected into the right substantia nigra compact and ventral tegmental area of rats to establish Parkinson’s disease models.The rats then received a transplantation of bone marrow stromal cells that were previously isolated,cultured and labeled with 5-bromo-2’-deoxyuridine in vitro.Transplantation of the bone marrow stromal cells significantly de-creased apomorphine-induced rotation time and the escape latency in the Morris water maze test as compared with rats with untreated Parkinson’s disease.Immunohistochemical staining showed that,5-bromo-2’-deoxyuridine-immunoreactive cells were present in the lateral ventricular wall and the choroid plexus 1 day after transplantation.These immunoreactive cells migrated to the surrounding areas of the lateral cerebral ventricle along the corpus callosum.The results indicated that bone marrow stromal cells could migrate to tissues surround the cerebral ventricle via the cerebrospinal fluid circulation and fuse with cells in the brain,thus altering the phenotype of cells or forming neuron-like cells or astrocytes capable of expressing neuron-specific proteins.Taken together,the present findings indicate that bone marrow stromal cells transplanted intracerebroventricularly could survive,migrate and significantly improve the rotational behavior and cognitive function of rats with experimentally induced Parkinson’s disease.     

Neuronal-like differentiation of bone marrow-derived mesenchymal stem cells induced by striatal extracts from a rat model of Parkinson’s disease

A rat model of Parkinson’s disease was established by 6-hydroxydopamine injection into the medial forebrain bundle. Bone marrow-derived mesenchymal stem cells (BMSCs) were isolated from the femur and tibia, and were co-cultured with 10% and 60% lesioned or intact striatal extracts. The results showed that when exposed to lesioned striatal extracts, BMSCs developed bipolar or multi-polar morphologies, and there was an increase in the percentage of cells that expressed glial fibrillary acidic protein (GFAP), nestin and neuron-specific enolase (NSE). Moreover, the percentage of NSE-positive cells increased with increasing concentrations of lesioned striatal extracts. However, intact striatal extracts only increased the percentage of GFAP-positive cells. The findings suggest that striatal extracts from Parkinson’s disease rats induce BMSCs to differentiate into neuronal-like cells in vitro.     

The role of DJ-1 in the pathogenesis of Parkinson’s disease

1 Introduction Parkinson’s disease (PD) is a common neurodegen- erative disorder characterized by the progressive loss of dopaminergic neurons in the substantia nigra pars com- pacta (SNpc). Although the majority of the cases appear to be sporadic, the disorder also can be associated with spe- cific genetic defects, several of which have been identified, including α-synuclein, parkin, PINK1, dardarin (LRRK2) and DJ-1[1]. Vincenzo Bonifati et al. localized a gene for autosomal recessive…     

伐仑克林靶向α-syn蛋白对帕金森病大鼠的机制研究

目的研究伐仑克林靶向α-syn蛋白对帕金森病(PD)大鼠的机制。方法选取SPF级30只SD健康大鼠,将30只大鼠随机分为空白组、模型组、用药组各10只,模型组和用药组建立PD大鼠模型。建模成功后,用药组使用1mg·kg-1的伐仑克林灌胃,空白组和模型组不做任何处理。在大鼠给药1w后对三组大鼠行为学、线粒体活性(State3、State4、RCR)、α-syn(α-突触核蛋白)、LPO(过氧化脂)、MDA(丙二醛)水平及对大脑组织中Beclin1、LC3B、APP合成酶活力、p-mTOR、IRE1α、ASK1、P-JNK蛋白表达量进行检测。结果模型组大鼠悬尾静止时间、IRE1α、ASK1、P-JNK蛋白表达量、α-syn、LPO、MDA水平均高于空白组,且糖水测试、Beclin1、LC3B蛋白表达量、线粒体活性、APP合成酶活力、p-mTOR均低于空白组,(P<0.05);用药组大鼠悬尾静止时间、IRE1α、ASK1、P-JNK蛋白表达量、α-syn、LPO、MDA水平均高于空白组,且糖水测试、Beclin1、LC3B蛋白表达量、线粒体活性、APP合成酶活力、p-mTOR均低于空白组,(P<0.05);用药组大鼠悬尾静止时间、IRE1α、ASK1、P-JNK蛋白表达量、α-syn、LPO、MDA水平均低于模型组,且糖水测试、Beclin1、LC3B蛋白表达量、线粒体活性、APP合成酶活力、p-mTOR均高于模型组(P<0.05)。结论伐仑克林通过靶向α-syn蛋白,作用于其线粒体功能及内质网应激机制,提高其行为学能力,效果显著。     

胚胎干细胞分化的神经前体细胞移植治疗帕金森病的实验研究

目的探讨胚胎干细胞（embryonic stemcell,ESC）来源的神经前体细胞（Neural precursorcell）移植治疗帕金森病的可能性。方法将胚胎干细胞诱导分化到神经前体细胞阶段后移植到大鼠帕金森病模型纹状体中,并设生理盐水组做对照研究,观察两组移植后行为学改变及检查纹状体内DA、DOPAC的含量。结果移植组在2～4周后与对照组相比行为学上有明显改善（P〈0．01）,纹状体内DA、DOPAC的含量显著提高（P〈0．01）。结论胚胎干细胞经诱导分化成神经前体细胞可用于帕金森病的修复治疗,胚胎干细胞是良好的干细胞移植治疗用细胞来源。     

止颤汤干预神经干细胞移植帕金森病大鼠脑内多巴胺及其代谢产物的变化*

背景：如何促进脑内多巴胺含量的增加以及减少多巴胺的代谢,是治疗帕金森病的热点所在。 目的：从多巴胺代谢途径角度观察止颤汤对神经干细胞移植帕金森病大鼠的脑黑质中多巴胺及其代谢产物含量的变化。 方法：以大鼠脑立体定位和1-甲基-4-苯基-1,2,3,6-四氢吡啶建立帕金森病大鼠模型。应用高效液相色谱法测定帕金森病大鼠中脑多巴胺及其代谢产物的含量。 结果与结论：止颤汤可以提高神经干细胞移植后帕金森病大鼠中脑多巴胺及其代谢产物双羟苯乙酸的含量,但对代谢产物高香草酸无明显影响。通过促进帕金森病大鼠干细胞移植后神经干细胞的存活,使之定向分化为多巴胺能神经元并分泌多巴胺,同时抑制多巴胺分解达到治疗作用。     

Gastrodin inhibits neuroinflammation in rotenone-induced Parkinson's disease model rats

The present study showed that the latency of rats moving on a vertical grid was significantly prolonged,and the number of rats sliding down from the declined plane was increased remarkably,in rotenone-induced Parkinson’s disease model rats compared with control rats.The moving latency recovered to normal levels,but the number of slides was significantly increased at 28 days after model establishment.The slope test is a meaningful approach to evaluate the symptoms of Parkinson’s disease model rats treated with rotenone.In addition,loss of substantia nigral dopaminergic neurons in model rats was observed at 1 day after the model was established,and continued gradually at 14 and 28 days.The expression of tyrosine hydroxylase-positive cells was significantly increased in gastrodin-treated rats at 14 days.Significant numbers of activated microglia cells were observed in model rats at 14 and 28 days;treatment of rats with Madopar at 28 days suppressed microglial activation.Treatment of rats with gastrodin or Madopar at 28 days significantly reduced interleukin-1β expression.The loss of substantia nigral dopaminergic neurons paralleled the microglial activation in Parkinson’s disease model rats treated with rotenone.The inflammatory factors tumor necrosis factor-α and interleukin-1β are involved in the substantia nigral damage.Gastrodin could protect dopaminergic neurons via inhibition of interleukin-1β expression and neuroinflammation in the substantia nigra.     

脐带间充质干细胞移植治疗帕金森病8例**

背景：中枢神经系统的神经细胞是一种终末细胞,任何病理过程导致的神经元丢失都将是一个不可逆过程。目的：观察间充质干细胞移植治疗帕金森病患者神经功能的效果。方法：选择2010-08/11上海455医院干细胞移植中心收治的帕金森病患者8例,男4例,女4例,年龄50~78岁。所有患者均住院治疗,自第2周起应用脐带间充质干细胞进行颈动脉穿刺移植治疗,采用帕金森病统一评分量表(Unified Parkinson's Disease Rating Scale,UPDRS)对患者移植前后神经功能进行评定,分值越高表示神经功能缺损越严重。结果与结论：8例患者均进入结果分析。与移植前相比,8例患者移植后1个月帕金森病统一评分量表分值均明显降低(P < 0.05),主要集中在对震颤、强直的改善,而运动迟缓、姿势不稳等临床症状无明显改善。8例患者均未出现移植物抗宿主病。提示脐带间充质干细胞移植可以一定程度地改善帕金森病患者的临床症状,提高患者生活质量。     

Stem cell transplantation for treating Parkinson's disease Literature analysis based on the Web of Science

OBJECTIVE:To identify global research trends of stem cell transplantation for treating Parkinson’s disease using a bibliometric analysis of the Web of Science.DATA RETRIEVAL:We performed a bibliometric analysis of data retrievals for stem cell transplantation for treating Parkinson’s disease from 2002 to 2011 using the Web of Science.SELECTION CRITERIA:Inclusion criteria:(a) peer-reviewed articles on stem cell transplantation for treating Parkinson’s disease which were published and indexed in the Web of Science;(b) type of articles:original research articles,reviews,meeting abstracts,proceedings papers,book chapters,editorial material and news items;(c) year of publication:2002-2011.Exclusion criteria:(a) articles that required manual searching or telephone access;(b) we excluded documents that were not published in the public domain;(c) we excluded a number of corrected papers from the total number of articles.MAIN OUTCOME MEASURES:(1) Type of literature;(2) annual publication output;(3) distribution according to journals;(4) distribution according to subject areas;(5) distribution according to country;(6) distribution according to institution;(7) comparison of countries that published the most papers on stem cell transplantation from different cell sources for treating Parkinson’s disease;(8) comparison of institutions that published the most papers on stem cell transplantation from different cell sources for treating Parkinson’s disease in the Web of Science from 2002 to 2011;(9) comparison of studies on stem cell transplantation from different cell sources for treating Parkinson’s disease RESULTS:In total,1 062 studies on stem cell transplantation for treating Parkinson’s disease appeared in the Web of Science from 2002 to 2011,almost one third of which were from American authors and institutes.The number of studies on stem cell transplantation for treating Parkinson’s disease had gradually increased over the past 10 years.Papers on stem cell transplantation for treating Parkinson’s disease appeared in journals such as Stem Cells and Experimental Neurology.Although the United States published more articles addressing neural stem cell and embryonic stem cell transplantation for treating Parkinson’s disease,China ranked first for articles published on bone marrow mesenchymal stem cell transplantation for treating Parkinson’s disease.CONCLUSION:From our analysis of the literature and research trends,we found that stem cell transplantation for treating Parkinson’s disease may offer further benefits in regenerative medicine.     

超顺磁氧化铁标记神经干细胞移植治疗帕金森病大鼠脑MRI改变的研究

目的研究应用超顺磁氧化铁（SPIO）标记神经干细胞（NSCs）移植治疗帕金森病（PD）大鼠脑部MRI的改变。方法从大鼠胚胎脑中分离培养中脑NSCs,用脂质体转染法将SPIO标记NSCs;同时制作大鼠PD模型,SPIO标记的NSCs移植到PD大鼠右侧纹状体区,分别在移植后1周、2周、4周、6周、8周和10周给大鼠行MRI检查,2周、4周、6周、8周、10周时进行旋转行为评分,并与NSCs组和对照组比较。结果脑内移植SPIO标记NSCs的PD大鼠1周后MRI检查显示在移植区呈低信号改变;移植后10周,在T2和T2梯度回波仍可观察到移植区的低信号,同时移植区有混杂信号。NSCs移植大鼠的旋转行为评分在移植后2-10周明显少于对照组（均P〈0.05）。结论SPIO标记的NSCs可以在MRI上显示其在移植大鼠脑部的分布和存活情况,有利于NSCs移植治疗PD后的疗效观察。