Research progress of neuroregeneration and multimodal neuromodulation in the treatment of spinal cord injury / 中国综合临床

Spinal cord injury has a high rate of disability in clinical practice, which can be divided into complete SCI and incomplete SCI according to different injury segments and severity.The main purpose of treatment is to protect the nerves.At present, acute spinal cord injury is mainly treated with surgical decompression, neurotrophic treatment, hormone therapy, hypothermia therapy, rehabilitation intervention and other clinical comprehensive treatment.In recent years, breakthroughs have been made in the field of endogenous and exogenous neural stem cell research, and important progress has been made in the basic research of stem cell transplantation.In the long run, nerve regeneration and nerve modulation may be the most promising therapy for the repair of spinal cord injury.

Development of electrical stimulation treatment in retina degeneration diseases / 国际眼科杂志(Guoji Yanke Zazhi)

@#Retinal diseases and optic nerve injury blocked visual signal transduction from retinal neurons to visual cortex, which would cause significant influence on patients' visual function and life quality. In clinic, glaucoma, traumatic optic neuropathy, and retinitis pigmentosa and so on are accompanied with degeneration of retinal neurons or optic nerve. However, efficient neuro-protective treatment is limited. Currently, studies suggested neuro-protective and regenerative effect of electrical stimulation treatment on retinal neurons and optic nerves. In this study, we reviewed the application of electrical stimulation in ophthalmology and summarized the possible mechanism, aiming to promote the development of electrical stimulation in the study and treatment of eye diseases.

Papel do receptor B2 de cininas na terapia da neurodegeneração dopaminérgica em modelo animal / Targeting Kinin-B2 receptors for the treatment of dopaminergic neurodegeneration in an animal mode

A Doença de Parkinson (DP) é um distúrbio neurodegenerativo, caracterizada em parte pela perda de neurônios dopaminérgicos da via nigroestriatal, originada na substância negra com projeções para o estriado, causando vários déficits motores. Atualmente, o tratamento mais utilizado é a administração de L-DOPA, um análogo da dopamina. Porém, essa droga apresenta eficácia limitada e induz diversos efeitos colaterais. A exploração dos efeitos neuroprotetores, proliferativos e neuroregenerativos da bradicinina (BK) em modelo animal de DP pode conduzir à substituição celular do tecido lesionado pela 6-hidroxidopamina (6-OHDA). De fato, a BK e seus receptores possuem um grande espectro de ações fisiológicas, estando classicamente envolvida no controle da homeostase cardiovascular e inflamação, além de exercer efeitos protetores em fisiopatologias do sistema nervoso, como em modelos de acidente vascular cerebral. Vários tipos celulares têm suas vias de sinalização associadas à ativação do receptor B2 de cininas (B2BKR). Trabalhos anteriores de nosso grupo mostraram que a BK está envolvida na diferenciação neural de células progenitoras neurais por um loop autócrino que resulta em ativação do B2BKR. Os resultados apresentados neste trabalho mostram a eficácia do tratamento com BK, um agonista de B2BKR, em animais submetidos à lesão da via nigro-estriatal induzida por 6-OHDA. Além disso, há uma recuperação comportamental e histológica desses animais quando tratados com Captopril®, um potencializador dos efeitos farmacológicos da BK, e com [Phe8Ψ(CH-NH)Arg9]-Bradicinina, agonista estável do receptor B2BKR. Assim, concluímos que a ativação de B2BKR pela BK desencadeiaum processo de neuroregeneração dopaminérgica de animais submetidos à lesão por 6-OHDA. Trabalhos recentes mostram que o receptor B2BKR desempenha um importante papel neuroprotetor em modelo animal da Doença de Alzheimer, o que corrobora nossos achados. Juntos, esses resultados contribuem para o estabelecimento da ação neuroprotetora e neurorregenerativa da BK no modelo de animal de neurodegeneração dopaminérgica, tornando-a uma excelente candidata para aplicação em terapias de reparo neuronal

Parkinson's disease (PD) is a neurodegenerative disorder partially characterized by the loss of dopaminergic neurons from the nigrostriatal pathway, originated in the substantia nigra with projections to the striatum, which causes several motor deficits. Currently, the most commonly used drug for PD treatment is levodopa. However, it has limited efficacy and induces several side effects. Elucidation of the neuroprotective, proliferative and neuroregenerative effects of bradykinin (BK) in animal models of PD can culminate in cellular replacement of the tissue damaged by 6-hydroxydopamine (6-OHDA). In fact, BK and its receptor have several physiological effects, being classically involved in the control of cardiovascular homeostasis and inflammation. Besides, BK exerts protective effects on nervous system pathophysiology, as observed in stroke models. Several cell types have their signaling pathways associated with the B2 kinin receptor (B2BKR) activation. Previous work from our group showed that BK is involved in differentiation of neural progenitor cells by an autocrine loop that results in activation of B2BKR. The results presented in this thesis show the efficacy of treatment with BK, through B2BKR activation, in animals submitted to nigrostriatal pathway injury induced by 6-OH dopamine. Furthermore, behavioral and histological recoveries of these animals were observed when treated with Captopril®, a potentiator of BK pharmacological effects, and with [Phe8Ψ (CH-NH) Arg9] -BK, a stable agonist of the B2BKR receptor. Thus, we conclude that BK activation of B2BKR triggers neuroregenerative processes in animals submitted to 6- OHDA injury. Recent studies showed that the B2BKR receptor plays an important neuroprotective role in an animal model of Alzheimer's disease, which corroboratesour findings. Together, these results contribute to the establishment of the neuroprotective and neuroregenerative actions of BK - an excellent candidate for neural repair therapies

Research progress in neuroregeneration of traumatic optic neuropathy / 中华实验眼科杂志

Traumatic optic neuropathy (TON) is commonly associated with blunt ocular trauma and craniofacial injuries,leading irreversible visual impairment.Like other mammalian mature central nerve system (CNS) neurons,retinal ganglion cell (RGC) is normally unable to regenerate axon spontaneously after optic nerve injury.The failure of axon regeneration has been attributed to the apoptosis of RGC,loss of intrinsic growth capacity of mature neurons,lack of suitable stimuli,and inhibition of extracellular environment.Via activating the intrinsic growth capacity of mature RGC,overcoming the inhibitory extracellular environment of damaged optic nerve,and providing appropriate inflammatory stimuli to initiate the regeneration process,mature RGC can be transformed into an active regenerative state allowing these neurons to survive axotomy and to regenerate axons in the injured optic nerve.Moreover,exploiting gene therapy based on adeno-associated virus may be a promising translatable treatment strategy for TON.

Effect of Enriched Environment on Neuroregeneration in Subventricle Zone of Rats with Vascular Dementia / 中国康复理论与实践

Objective To explore the effect of enriched environment on neuroregeneration in subventricle zone(SVZ)of rats with vascu-lar dementia.Methods All rats were divided into sham group(n=5),vascular dementia group(VD group,n=12)and enriched environment group(EE group,n=12).Then,2-VO method was applied to make the vascular dementia model.The sham group and VD group received conventional breeding environment for 30 days,while EE group was subjected to enriched environment for 30 days.Immunohistochemistry was applied to detect the BrdU and Doublecortin(DCX)expression in SVZ,and DCX/Ki67,DCX/p-cAMP-response element binding pro-tein(CREB)expression in SVZ was assessed by immunofluorescence double-labeling technique.Results Compared with the sham group, the number of DCX+cells(t=2.989,P=0.026)and BrdU+cells(t=3.069,P=0.005)increased in VD group,and increased more in EE group (t=3.067, P=0.027; t=2.907, P=0.011). Besides, the number of the DCX/Ki67 (t=2.994, P=0.040) and DCX/p-CREB (t=4.707, P=0.009) cells was significantly higher in EE group than in VD group.Conclusion Enriched environment could up-regulate the neuroregeneration ca-pacity in SVZ of rats with vascular dementia through CREB signal pathway.

Promoting effect of percutaneous electrical stimulation on nerve regeneration after end-to-side neurorrhaphy in rats / 中国比较医学杂志

Objective To observe and explore the effect and clinical value of percutaneous electrical stimulation on nerve regeneration after end-to-side neurorrhaphy in rats. Methods Thirty-two SPF male S-D rats were randomly divided into four groups ( n = 8 ): group A, the normal control group; group B, with end to end neurorrhaphy of musculocutaneous nerve injury matched to the ulnar nerve; group C, with end to side neurorrhaphy of musculocutaneous nerve injury matched to the ulnar group; and group D, with end to side neurorrhaphy of musculocutaneous nerve injury matched to the ulnar nerve plus postoperative transcutaneous electrical stimulation ( 30 min per day for 6 weeks ) . Electromyography, postoperational nerve conduction velocity, the histological and ultrastructural changes of the nerve fibers were examined, and NF-200 expression in frozen sections was observed using imunohistological staining, to assess the recovery of muscle strength of the diseased side limb and the neuroregeneration in the rats after treatment. Results The amplitude and conduction velocity of the groups C and D were lower than that of the group A, the latency was higher than that of the group A, while the amplitude and conduction velocity of the group D were lower than that of the group C,and the latency was higher than that of the group C. The wet weight ratio of the biceps brachii muscle and the cross-sectional area of muscle fibers in the groups B, C and D were lower than those in the group A, and the recovery of muscle in the group C was the worst. The expression of NF-200 in the rats of groups B, C and D was significantly lower than that in the group A, and the expression of NF-200 in the group D was significantly higher than that in the group C, but still significantly less than that in the group B ( P < 0. 05 ) . Electron microscopy showed mature myelinated fibers in the group B, whereas unmyelinated fibers were the main component and the myelin sheath was poorly developed in the group C. The myelin regeneration in the group D was better than that in the group C, but still some unmyelinated nerve fibers were seen. Conclusions The percutaneous electrical stimulation can effectively promote nerve axonal regeneration and can delay the atrophy of the target muscle after end-to-side neurorrhaphy. Though there is difference compared with the end-to-end neurorrhaphy, the end-to-side neurorrhaphy is still an effective method in clinical repair of peripheral nerve injury.

Effect of total saponins of Panax notoginseng on expression of GFAP in hippocampus and brain water content in rats subjected global cerebral ischemia injury / 中草药

Objective To explore the effect of the total saponins of Panax notoginseng (TSPN) on the expression of GFAP in hippocampus and brain water content in rats subjected global cerebral ischemia injury. Methods Using four-vessel occlusion method built the global cerebral ischemia model. Rats were divided into Sham group, vehicle group, and TSPN group. The rats in the TSPN group were administered TSPN intraperitoneally 30 min post-brain ischemia. The dose of TSPN (75 mg/kg) was suspended in 0.9% saline, once per day for days 1, 3, 7, and 14 after reperfusion. While rats in the vehicle group was treated with equal volume of 0.9% saline, one injection per day until the rats were sacrificed at either days 1, 3, 7, and 14 after brain ischemia. The brain water contentwas detected by dry-wet technique and GFAP expression in the dentate subgranular zone (SGZ) was assessed by immunohistochemistry. Moreover, immunofluorescence was applied to detect the GFAP/DCX in the SGZ, and western-blot was adopted to test the protein level of GFAP. Results The brain water content in the TSPN group was significantly lower than the vehicle group (P < 0.05); There was statistical difference in the GFAP+ cells density in SGZ at the 3th, 7th, 14th day in two groups (P < 0.05). Furthermore, compared with the vehicle group, the ratio of the GFAP/DCX to DCX in the SGZ at 7th, 14th d of TSPN group was significantly different (P < 0.05), and the protein level of GFAP on days 3, 7, 14 in the TSPN group was higher (P < 0.05). Conclusion TSPN could play a neuroprotective effect through promoting gliosis, neuroregeneration in the SGZ, and alleviating brain water content of rats following global cerebral ischemia.

Effect of total saponins of Panax notoginseng on neuroregeneration in subventricle zone of rats with global cerebral ischemia / 中草药

Objective: To explore the effect of total saponins of Panax notoginseng (TSPN) on the neuroregeneration in subventricle zone (SVZ) in rats with global cerebral ischemia. Methods: Using four-vessel occlusion method to build the global cerebral ischemia model. Rats were divided into Sham group, vehicle group, and TSPN group. The rats in TSPN group were ip administered with TSPN 30 min post-brain ischemia. The dose of TSPN (75 mg/kg) was suspended in 0.9% saline (10 g/L), once per day for 1, 3, 7, 14 days after reperfusion. While rats in the vehicle group were treated with equal volume of 0.9% saline, one injection per day until the rats were sacrificed at either 1, 3, 7, and 14 days after brain ischemia. The BrdU and Doublecortin (DCX) expression in SVZ was assessed by immunohistochemistry and applying the immunofluorescence double-labelling to detect the BrdU/DCX, DCX/Ki67, and GFAP/DCX in SVZ. Results: In comparison with the vehicle group, the number of BrdU+ cells in SVZ of TSPN group was significantly higher on days 7 and 14 (P < 0.01, 0.001); Statistical meaning existed in two groups on days 7 and 14 about the mean optical density of DCX+ cells in SVZ (P < 0.01, 0.001). In comparison with the vehicle group, the number of the BrdU-labeled cells co-expressing DCX in the SVZ on day 14t of TSPN group was significantly different (P < 0.01, 0.001). There was statistical meaning in comparison of the number of colocalization of DCX with Ki67 on days 7 and 14 in SVZ between the TSPN group and vehicle group (P < 0.01, 0.001). The ratio of GFAP/DCX to DCX in SVZ of two groups were statistically different on days 2, 7, and 14 (P < 0.05, 0.001). Conclusion: TSPN could promote the neuroregeneration, drive the proliferation and differentiation of neural progenitor cells, and enhance the differentiation of gliosis into newborn immature neurons in SVZ of rats with global cerebral ischemia.

The Therapeutic Potential of Stem Cells and Progenitor Cells for the Treatment of Parkinson's Disease

Parkinson's disease (PD) is the second most common neurodegenerative disorder. It is usually seen in those above 50 years old. Current medical treatments only provide symptomatic relief but cannot cure the disease. There are claims that PD can be cured by stem cell transplant. The present study is aimed to assess the clinical potency and safety of stem cell in treating PD. A total of eleven articles were included for analysis, with four randomised control trials (RCTs), five non-RCTs and 2 follow up studies. All the four non-RCTs showed improvement of Unified Parkinson's Disease Rating Scale with no adverse events. However, results from RCTs showed no significant differences in the rating score among the transplant group and the Sham surgery group. The secondary analysis of one study showed a significant improvement of the rating score in those patients aged 60 and younger. Transplant group also associated with an overall higher incidence of adverse events. In conclusion, the RCTs and non-RCTs produced opposite results. When the studies were performed as non-RCTs in small number of patients, they showed promising result in the patients. It could say that currently the use of stem cell/progenitor cells in treating PD need much research despite having the implanted stem cell to be able to survive and integrated. The survival of implanted dopamine neurons in the striatum, however, does not indicate a success in correcting PD symptoms. Further investigations will shed light on the application and mechanism of action of stem cells in treating PD.

Morphological characterization of gecko's (eublepharis macularius) glial cells in culture / Caracterización morfológica de las células gliales del gecko (eublepharis macularius) en cultivo

Central nervous system of reptiles has the ability to grow and regenerate during adult life of the animal. Therefore, cells creating CNS of this animal class should compound substances or molecules enabling neuroregeneration. Cells directly involved in this process have not been clearly characterized, especially in cell culture environment. Morphology of reptilian glial adherent cells should be known better to find any differences from mammalian CNS cells. We isolated glial cells from olfactory bulb and cerebrum from gecko (Eublepharis macularius) and cultured separately. We have observed populations of cells with proliferative capacity in both types of cultures. Also, we have detected lipid molecules deposits within their cytoplasm, which localization was correlated with mitochondria position. This information can be helpful in searching new bioactive substances involved in regeneration of central nervous system.

El sistema nervioso central de los reptiles tiene la capacidad de crecer y regenerarse durante la vida adulta del animal. Por lo tanto, las células de SNC creadas de esta clase de animales deberían componerse de sustancias o moléculas que permiten la neuroregeneración. Las células que participan directamente en este proceso no han sido claramente caracterizadas, especialmente en el entorno de cultivo celular. La morfología de las células adherentes gliales de reptiles deben ser reconocidas y diferenciarse respecto a las células del SNC de mamíferos. Se aislaron células gliales del bulbo olfatorio y el cerebro del Gecko (Eublepharis macularius) y se cultivaron por separado. Se observaron poblaciones de células con capacidad proliferativa en ambos tipos de cultivos. Además, se detectaron moléculas de depósitos lipídicos dentro de su citoplasma, y su localización se correlacionó con la posición de las mitocondrias. Esta información puede ser útil en la búsqueda de nuevas sustancias bioactivas que participan en la regeneración del sistema nervioso central.

Efficacy and Tolerability of MlC601 in Patients with Mild to Moderate Alzheimer Disease who were Unable to Tolerate or Failed to Benefit from Treatment with Rivastigmine.

Aim: To evaluate the efficacy and tolerability of MLC601 in patients with mild to moderate Alzheimer disease (AD). Study Design: This is an open-label pilot study. Place and Duration of Study: It was conducted at three university referral centres in Iran from September 2009 until November 2011. Methodology: One-hundred and twenty four outpatients with mild to moderate AD who had previously failed to tolerate or benefit from treatment with Rivastigmine for 6 months at a dose of 2 to 12 mg per day were switched to a MLC601 regimen of one capsule three times per day for up to 18 months. Outcome measures included adverse events (AEs), withdrawal rate, and changes in the Mini-Mental State Examination (MMSE) and the cognitive subscale of the AD Assessment Scale (ADAS-cog) relative to baseline measurements. Results: Two patients were lost to follow up, and 122 patients completed the 18-month trial. The mean age of the participants was 65.3±6.4 years (range 54-82), and 77 (63.1%) of the participants were female. Improved cognitive function was observed in the first 6 months of the regimen (ADAS-cog=-3.1±10.1; MMSE=1.2±3.0), and the stabilisation of cognitive decline was observed over the remaining 12 months (ADAScog=- 1.6±7.6; MMSE=0.8±4.2). AEs were predominantly gastrointestinal and occurred in 7.3% of patients. Conclusions: MLC601 showed good tolerability and promising effects on cognitive function in AD patients during 18 months of treatment.

Olfactory tract transection reveals robust tissue-level plasticity by cellular numbers and neurotrophic factor expression in olfactory bulb.

Nervous system lesions are characterized by the loss of neuronal numbers and types. The neurotrophic factor levels in an injured tissue reflect their potential for regeneration. This hypothesis was investigated in olfactory bulb (OB), where olfactory tract was surgically transected disrupting neuronal migration and turnover. The effects were followed with quantification of mitral cells and three neurotrophic factors mRNA levels for 6 weeks. The neuronal numbers decreased by 3rd- and 4th-week in transected OBs followed by their restoration, comparable with that of controls at 5th- and 6th-week. The endogenous levels of three neurotrophic factors – (brain derived neurotrophic factor, insulin growth factor-1 and fibroblast growth factor-2) using qPCR showed increase at 2nd-week by 136-, 8- and 2-fold respectively. Also, there was a significant increase in specific neurotrophic factors at 5th-week and 6th-weeks. The results propose a temporal link between deployment of neurotrophic factors and the plausible restorative events for mitral cell numbers in OB.

Autologous Bone Marrow Stem Cell Therapy Shows Functional Improvement in Hemorrhagic Stroke.

In hemorrhagic stroke, damage to the brain tissue is inevitable and no effective treatment for functional improvement is currently available except neurorehabilitation. Stem cell therapy is a rapidly growing field and has recently opened new avenues for brain repair strategies. We present a case study of a 69-year-old female treated with stem cell therapy for right-sided hemiplegia caused due to left thalamic hemorrhagic stroke. Inspite of regular physiotherapy, the patient had constant residual neurodeficit, one year after the stroke, which was severely incapacitating. In view of the same, the patient was given intrathecal autologous bone marrow derived stem cell therapy as part of the neuroregeneration and rehabilitation therapy (NRRT) along with rehabilitation. After the therapy, patient showed functional as well as neurological improvements (cognition and motor strength) without any side effects. There is accumulating experimental data showing the benefits of cell transplantation on functional recovery after hemorrhagic stroke. This case study supports the concept of neuroregeneration with bone marrow stem cells as a novel strategy having great therapeutic potential. However, large clinical studies are needed to further investigate autologous bone marrow stem cell therapy in addition to neurorehabilitation for treating the disability in hemorrhagic stroke.

Pantothenate Kinase Associated Neurodegeneration: A Case Report.

Pantothenate kinase-associated neurodegeneration (PKAN) is a form of neurodegeneration with brain iron accumulation (NBIA), formerly called Hallervorden-Spatz syndrome. PKAN is the first described inborn error of coenzyme A metabolism. PKAN encompasses two clinical subtypes, classic and atypical. We here report an eight year girl diagnosed as classic PKAN. We also review the literature about PKAN.

Differential astroglial responses in the spinal cord of rats submitted to a sciatic nerve double crush treated with local injection of cultured Schwann cell suspension or lesioned spinal cord extract: implications on cell therapy for nerve repair

PURPOSE: Reactive astrocytes are implicated in several mechanisms after central or peripheral nervous system lesion, including neuroprotection, neuronal sprouting, neurotransmission and neuropathic pain. Schwann cells (SC), a peripheral glia, also react after nerve lesion favoring wound/repair, fiber outgrowth and neuronal regeneration. We investigated herein whether cell therapy for repair of lesioned sciatic nerve may change the pattern of astroglial activation in the spinal cord ventral or dorsal horn of the rat. METHODS: Injections of a cultured SC suspension or a lesioned spinal cord homogenized extract were made in a reservoir promoted by a contiguous double crush of the rat sciatic nerve. Local injection of phosphate buffered saline (PBS) served as control. One week later, rats were euthanized and spinal cord astrocytes were labeled by immunohistochemistry and quantified by means of quantitative image analysis. RESULTS: In the ipsilateral ventral horn, slight astroglial activations were seen after PBS or SC injections, however, a substantial activation was achieved after cord extract injection in the sciatic nerve reservoir. Moreover, SC suspension and cord extract injections were able to promote astroglial reaction in the spinal cord dorsal horn bilaterally. Conclusion: Spinal cord astrocytes react according to repair processes of axotomized nerve, which may influence the functional outcome. The event should be considered during the neurosurgery strategies.

OBJETIVO: Astrócitos reativos participam de vários mecanismos após lesões do sistema nervoso central e periférico, os quais incluem neuroproteção, brotamento neuronal, neurotransmissão e dor neuropática. As células de Schwann (CS), um tipo de glia periférica, também reagem com a lesão do nervo, podendo interferir com o reparo e cicatrização, crescimento de fibras e regeneração neuronais. Investigamos aqui a possibilidade da terapia celular para o reparo do nervo ciático poder alterar o padrão da ativação astrocitária nos cornos anterior e posterior da medula espinal do rato. MÉTODOS: Suspensão de CS cultivadas ou extrato homogeneizado de medula espinal lesada de rato foram inoculados num reservatório feito a partir de dois esmagamentos aplicados no nervo ciático do rato distantes 0,5mm entre si. Injeção local de salina tamponada serviu como controle. Os ratos foram mortos uma semana após e os astrócitos da medula espinal marcados por método imunohistoquímico e quantificados por análise de imagem. RESULTADOS: No corno anterior da medula, ipsilateral à lesão, ativação astrocitária leve foi vista após as injeções de tampão ou CS, entretanto, ativação celular intensa foi observada nesta região com a inoculação neural do extrato homogeneizado de tecido medular lesado. Adicionalmente, as inoculações de CS e de extrato homogeneizado de tecido medular promoveram forte reação astrocitária no corno dorsal da medula espinal, bilateralmente. CONCLUSÕES: Os astrócitos da medula espinal reagem em função do processo de reparo do nervo lesado, o que pode influenciar o resultado funcional esperado, algo que deve ser considerado durante o planejamento da estratégia neurocirúrgica.

THE ESTABLISHMENT OF MODEL FOR PRIMARY CULTURED TANYCYTES / 解剖学报

Objective Tanycytes(TAs) is a specialized eppendymal glia that locates mostly in the ventral lateral wall of the ventricle III and median eminence(ME).Due to its peculiar location and directly exposure to the cerebrospinal fluid,blood,neuroendocrine hormones and neurons,tanycytes play an important role in the brain barrier system,brain-CSF neurohumoral circuit and immune-neuroendocrine network.They maintain immature characters during the adulthood and have naturally conducted the neuroregeneration process in the adult hypothalamus of mammal animals.This research was designed to identify tanycytes(TAs) in postnatal 7 days of Wistar rats and then establish the cell model of TAs for further study.Methods Tanycytes of postnatal 7 days of Wistar rats were identified by the immunohistochemical technology using glial fibrillary acidic protein(GFAP) and vimentin(VIM),which are the intermediate filament markers of glia cells.The qualitative analysis was performed by the NADPH-d stain of nitric oxide sythenase(NOS).Primary TAs model was established by the cell culture technique and identified by the same methods as in vivo.Results Primary cultured TAs expressed VIM,GFAP and NOS which was identical with those in vivo.Conclusion Cell model of TAs from postnatal 7days of Wistar rats in vitro has been established and TAs could be used as a proper substrate for transplanting.