血清和雪旺氏细胞诱导大鼠胚胎神经干细胞分化的比较

目的 比较血清和雪旺氏细胞诱导大鼠胚胎神经干细胞分化的差异。方法 分别采用血清和与雪旺氏细胞共培养的方法诱导大鼠胚胎神经干细胞分化，应用相差显微镜和免疫荧光染色的方法对其进行观察和比较。结果 两种方法都能够诱导绝大多数神经干细胞分化成神经元，少量分化成星形胶质细胞和少突胶质细胞。虽然后一种方法诱导干细胞分化的进程比前一种方法要慢，但细胞形态学上更接近发育成熟的神经元。结论 雪旺氏细胞的分泌物不仅能够诱导共培养的神经干细胞分化，而且使其分化更加成熟。     

Human umbilical cord Wharton's jelly-derived mesenchymal stem cells differentiate into a Schwann-cell phenotype and promote neurite outgrowth in vitro

Cell-based therapy has achieved promising functional recovery for peripheral nerve repair. Although Schwann cells (SCs) and bone marrow derived mesenchymal stromal cells (BM-MSCs) are the main cell source for nerve tissue engineering, the clinical application is limited because of donor site morbidity, the invasive procedure, and the decreased number of SCs and BM-MSCs. Wharton's jelly-derived mesenchymal stem cells (WJMSCs) could be a promising cell source for nerve tissue engineering because they are easily accessible and their use has no ethical issues. We investigated the phenotypic, molecular and functional characteristics of WJMSCs differentiated along a Schwann-cell lineage. Cultured WJMSCs were isolated from human umbilical cord, and the undifferentiated WJMSCs were confirmed by the detection of MSC-specific cell-surface markers. WJMSCs treated with a mixture of glial growth factors (basic fibroblast growth factor, platelet-derived growth factor and forskolin) adopted a spindle-like morphology similar to SCs. Immunocytochemical staining, RT-PCR analysis, and Western blot analysis revealed that the treated cells expressed the glial markers glial fibrillary acidic protein, p75, S100 and P0 and indicative of differentiation. On co-culture with dorsal root ganglia neurons, the differentiated WJMSCs enhanced the number of sprouting neurites and neurite length in dorsal root ganglia neurons. Furthermore, using enzyme-linked immunosorbent assay and RT-PCR methodology, we found differentiated WJMSCs secrete and express neurotrophic factors, including brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), and neurotrophin-3 (NT-3). Quantification of neurite outgrowth from PC12 cells grown in differentiated WJMSCs-conditioned media demonstrates that the neurite length is significantly more than control medium and undifferentiated WJMSCs group. WJMSCs can be differentiated into cells that are Schwann-like in terms of morphologic features, phenotype, and function and could be suitable Schwann-cell substitutes for nerve repair in clinical applications.     

骨髓基质细胞体外分离培养和分化为

摘要 目的：观察体外培养大鼠骨髓基质细胞（BMSCs bone marrow stromal cells）的生物学特性并探讨其分化为Schwann细胞表达S100的机制,为组织工程学修复周围神经损伤奠定一定的实验基础。方法：从成年SD大鼠的股骨和胫骨中分离培养BMSCs,倒置显微镜下动态观察细胞的生长状态,采用MTT法检测细胞的活力,FCM检测细胞周期等方法研究BMSCs的生物学特性。应用复合诱导因子（BME, RA ,FSK, bFGF, PDGF, HRG）体外诱导BMSCs向Schwann细胞分化。诱导分化后采用免疫荧光细胞化学染色,流式细胞仪,逆转录PCR方法分别检测检测胶质细胞标记蛋白S100蛋白和mRNA的表达。结果：BMSCs在体外容易扩增,传1－8代以内的细胞增殖能力无明显变化。未经诱导的BMSCs大部分处于G0和G1期。诱导分化后的细胞形态上类似Schwann细胞,并且表达胶质细胞的标记蛋白S100蛋白及其mRNA明显升高。结论：BMSCs在体外可以向类Schwann细胞诱导分化,并且表达胶质细胞的标记蛋白S-100蛋白及其mRNA。     

体外诱导获得雪旺细胞的可靠性研究

目的 研究骨髓基质细胞(BMSCs)在体外条件下向周围神经雪旰细胞(SCs)分化的可靠性. 方法 分离提取SD大鼠股骨和胫骨部位BMSCs.利用其贴壁生长的特性.培养纯化,传代扩增.用复合诱导因子(β.巯基乙醇+全反式维甲酸+血小板凝集抑制剂+m小板源性生长因子+碱性成纤维细胞生长因子)在体外诱导BMSCs分化.免疫细胞化学方法 检测P75、S-100及胶质原纤维酸性蛋白(GFAP)的表达,荧光实时定量PCR检测P75、S100及CD104的表达. 结果诱导后的BMSCs形态类似SCs,免疫荧光染色鉴定其具有SCs性质,表达SCs的表面标志物(GFAP、S100和P75).荧光实时定量PCR结果显示诱导后BMSCs S-100、CD104的表达量达到了SCs的表达量水平,但P75的表达量与SCs的表达量水平还有较大差距. 结论 体外诱导BMSCs可部分获得SCs的特征,传代后恢复至未诱导状态,这种预诱导加复合因子诱导的方法 尚待完善.     

星形胶质细胞诱导成年骨髓基质细胞分化成神经元样细胞的实验研究

目的探讨星形胶质细胞能否在体外诱导成年大鼠骨髓基质细胞向神经元方向分化。方法采用新生鼠海马组织来源的星形胶质细胞与转染有绿色荧光蛋白基因的骨髓基质细胞(MSCs)进行共培养,并分成三组:共培养组、共培养 脑源性神经营养因子(BDNF)组、单纯碱性成纤维生长因子(bFGF) BDNF诱导分化组,在相差显微镜下每日观察、记录MSCs的诱导分化状况,并应用免疫荧光染色技术对分化后的MSCs进行鉴定,同时应用流式细胞术测定MSCs分化前后及星形胶质细胞的DNA含量。结果共培养第4天,部分MSCs已初步具备神经元形态:折光性强的锥形或圆形胞体及长的多极突起,免疫荧光染色呈微管相关蛋白(MAP-2ab)、神经元特异性烯醇化酶(NSE)阳性;DNA的含量测定结果显示诱导分化后第7天未发现有四倍体、六倍体细胞。结论(1)星形胶质细胞可以在体外诱导成年大鼠MSCs向神经元方向分化,这种分化并不是由细胞融合引起的。(2)星形胶质细胞具有调控神经元的分化、促进神经元成熟的功能。     

Differentiation of rat adipose tissue-derived stem cells into Schwann-like cells in vitro

In this study, we explored the competence of adipose-derived stem cells to differentiate into Schwann cells in vitro. Rat adipose-derived stem cells were sequentially treated with various factors beta-mercaptoethanol, all-trans-retinoic acid, followed by a mixture of forskolin, basic fibroblast growth factor, platelet-derived growth factor and heregulin. We found that differentiated adipose-derived stem cells displayed the morphology of Schwann cells. Western blotting and dual immunofluorescence staining confirmed that they produced proteins characteristic for Schwann cells, including S100 and glial fibrillary acidic protein. Furthermore, differentiated adipose-derived stem cells could enhance neurite outgrowth in coculture with sensory neurons. These results demonstrate that adipose-derived stem cells can differentiate into Schwann-like cells with morphological, phenotypic, and functional characteristics of Schwann cells.     

Differentiation of mesenchymal stem cells to support peripheral nerve regeneration in a rat model

Mesenchymal stem cells (MSCs) support axon regeneration across artificial nerve bridges but their differentiative capacity and ability to promote nerve regeneration remains unclear. In this study, MSCs isolated from bone marrow of Sprague–Dawley rats were characterized by plastic adherence and pluripotency towards mesodermal lineages. Isolated undifferentiated MSCs (uMSCs) were stimulated towards a Schwann cell (SC) phenotype using specific growth factors, and cell marker analysis was performed to verify SC phenotype in vitro. Differentiation resulted in temporally dependent positive immunocytochemical staining for the SC markers, glial fibrillary acidic protein (GFAP), S100, and nerve growth factor receptor (NGFR), with maximal marker expression achieved after 6 days of treatment with differentiation media. Quantitative analysis demonstrated that ~ 50% of differentiated MSCs (dMSCs) have a SC phenotype. Using an indirect co-culture system, we compared the ability of dorsal root ganglion (DRG) cells to extend neurites in indirect contact with uMSCs and dMSCs as compared to SCs. The mean values of the longest length of the DRG neurites were the same for the dMSCs and SCs and significantly higher than the uMSC and DRG mono-culture systems (p < 0.05). In vivo, compared to an empty conduit, dMSC seeded collagen nerve conduits resulted in a greater number of sciatic motoneurons regenerating axons through the conduit into the distal nerve stump. We conclude that bone marrow-derived MSCs differentiate into a SC-phenotype that expresses SC markers transiently and sufficiently to support limited neurite outgrowth in vitro and axonal regeneration equivalent to that of SCs in vitro and in vivo. The nerve autograft remains the most effective conduit for supporting regeneration across nerve gaps.     

Mesenchymal stem cells expressing neural antigens instruct a neurogenic cell fate on neural stem cells

The neurogenic response to injury in the postnatal brain is limited and insufficient for restoration of function. Recent evidence suggests that transplantation of mesenchymal stem cells (MSCs) into the injured brain is associated with improved functional recovery, mediated in part through amplification in the endogenous neurogenic response to injury. In the current study we investigate the interactions between bone marrow-derived MSCs and embryonic neural stem cells (NSCs) plus their differentiated progeny using an in vitro co-culture system. Two populations of MSCs were used, MSCs induced to express neural antigens (nestin+, Tuj-1+, GFAP+) and neural antigen negative MSCs. Following co-culture of induced MSCs with differentiating NSC/progenitor cells a significant increase in Tuj-1+ neurons was detected compared to co-cultures of non-induced MSCs in which an increase in astrocyte (GFAP+) differentiation was observed. The effect was mediated by soluble interactions between the two cell populations and was independent of any effect on cell death and proliferation. Induced and non-induced MSCs also promoted the survival of Tuj-1+ cell progeny in long-term cultures and both promoted axonal growth, an effect also seen in differentiating neuroblastoma cells. Therefore, MSCs provide instructive signals that are able to direct the differentiation of NSCs and promote axonal development in neuronal progeny. The data indicates that the nature of MSC derived signals is dependent not only on their microenvironment but on the developmental status of the MSCs. Pre-manipulation of MSCs prior to transplantation in vivo may be an effective means of enhancing the endogenous neurogenic response to injury.     

Sciatic nerve regeneration in rats induced by transplantation of in vitro differentiated bone-marrow stromal cells

Bone marrow stromal cells (MSCs) are multipotent stem cells that have the potential to differentiate into bone, cartilage, fat and muscle. We now demonstrate that MSCs can be induced to differentiate into cells with Schwann cell characteristics, capable of eliciting peripheral nervous system regeneration in adult rats. MSCs treated with beta-mercaptoethanol followed by retinoic acid and cultured in the presence of forskolin, basic-FGF, PDGF and heregulin, changed morphologically into cells resembling primary cultured Schwann cells and expressing p75, S-100, GFAP and O4. The MSCs were genetically engineered by transduction with retrovirus encoding green fluorescent protein (GFP), and then differentiated by treatment with factors described above. They were transplanted into the cut ends of sciatic nerves, which then responded with vigorous nerve fibre regeneration within 3 weeks of the operation. Myelination of regenerated fibers by GFP-expressing MSCs was recognized using confocal and immunoelectron microscopy. The results suggest that MSCs are able to differentiate into myelinating cells, capable of supporting nerve fibre re-growth, and they can therefore be applied to induce nerve regeneration.     

脐血源性神经球分化为类雪旺细胞的研究

目的 研究脐血间充质干细胞(CB-MSC)向雪旺细胞谱系分化的生物表型和特征.方法诱导CB-MSC为悬浮的神经球,然后用胶质生长因子诱导分化为类雪旺细胞.分化的CB-MSC表现出和雪旺细胞类似的形态学的改变.通过免疫化学染色和Western blotting鉴定这些细胞雪旺细胞标志的表达情况;通过分化的CB-MSC与背根神经节神经元共培养观察其促进其轴突生长的作用.结果 未分化的CB-MSC很少表达nestin、GFAP和S-100.在CB-MSC诱导成神经球以后,82.38%±3.70%的细胞表达nestin,而GFAP和S-100仍然阴性.接种于分化液36 h后细胞表达nestin、GFAP、S-100,阳性率分别为43.78%±3.21%、35.42%±1.82%和29.49%±2.54%.相比未分化CB-MSC,分化的CB-MSC中神经轴突的形成率和轴突的长度明显增加,差异有统计学意义(P＜0.05).结论CB-MSC可以通过诱导转变为神经球,神经球可以诱导成雪旺细胞的形态,表型和功能类似雪旺细胞,可做为一种替代骨髓间充质干细胞的细胞来源,对于治疗神经系统疾病具有潜在的应用价值.     

Anatomical site influences the differentiation of adipose-derived stem cells for Schwann-cell phenotype and function

Considerable attention has recently been given to adipose-derived stem cells (ASCs) as an important source for differentiation to Schwann cells in the treatment of peripheral nerve injury, with considerable clinical advantages over the use of mesenchymal stem cells derived from bone marrow or autologous Schwann cells. However, the relationship between adipose donor site and differentiated ASC phenotype and function is presently unknown. This work systematically studied the differentiation of ASCs harvested from three anatomical sites: (i) subcutaneous; (ii) perinephric; and (iii) epididymal adipose tissue. We show that ASC source is a major determining factor of immunophenotype, multilineage differentiation, Schwann-cell protein expression, and paracrine ability to stimulate neuronal growth. Upregulation of S100β, glial fibrillary acidic protein (GFAP), and p75NGFR was observed in differentiated ASCs from perinephric fat tissue, while only the expression of S100β or GFAP and p75NGFR was elevated in differentiated ASCs from subcutaneous or epididymal fat tissue. Although the co-culture of differentiated ASCs with NG108-15 neuronal cells demonstrated that ASCs from each source could stimulate neurite outgrowth and number, differentiated ASCs from subcutaneous and perinephric fat versus epididymal fat were most effective, which was attributed to high-brain-derived neurotropic factor/nerve growth factor and low-neurotrophin-3 levels. Thus, ASCs can be obtained from different anatomical locations, and this determines Schwann-cell phenotype upon differentiation and extent of function. This work is therefore of relevance in local therapeutic delivery of ASCs for the repair of peripheral nerve injury, but also in the broader context of ASC use in related stem-cell therapies.     

Fibroblast growth factor-20 promotes the differentiation of Nurr1-overexpressing neural stem cells into tyrosine hydroxylase-positive neurons

Stem cells are currently considered as alternative cell resources for restorative transplantation strategies in Parkinson's disease. However, the mechanisms that induce differentiation of a stem cell toward the dopaminergic phenotype are still partly unknown thus hampering the production of dopaminergic neurons from stem cells. In the past, FGF-20 has been found to promote the survival of ventral mesencephalic (VM) dopaminergic (DA) neurons in culture. We hereby provide evidence that FGF-20, a growth factor of the FGF family, is expressed in the adult and 6-OHDA-lesioned striatum and substantia nigra, but is not expressed by VM glia or DA neurons, suggesting that FGF-20 may work on DA neurons in a paracrine- or target-derived manner. We also found that co-culture of Nurr1-NSCs with Schwann cells overexpressing FGF-20 induced the acquisition of a neuronal morphology by the NSCs and the expression of tyrosine hydroxylase (TH) as assessed by immunocytochemistry, cell ELISA, and Western blot analysis. RT-PCR showed, that both, Schwann cells and Nurr1-NSCs (differentiated or not), expressed the FGF-1 receptor suggesting that both direct and indirect actions of FGF-20 are possible. We show that differentiated Nurr1 cells retained both neuronal morphology and TH expression after transplantation into the striatum of 6-OHDA-lesioned postnatal or adult rats, but that neuritogenesis was only observed after postnatal grafts. Thus, our results suggest that FGF-20 promotes the differentiation of Nurr1-NSCs into TH-positive neurons and that additional factors are required for the efficient differentiation of DA neurons in the adult brain.     

Bone marrow-derived Schwann cells achieve fate commitment – a prerequisite for remyelination therapy

Schwann cell transplantation improves post-traumatic nerve regeneration in both PNS and CNS but sufficient numbers of immunocompatible cells are required for clinical application. Currently, Schwann cell-like cells derived from the bone marrow lack fate commitment and revert to a fibroblast-like phenotype upon withdrawal of differentiation-inducing factors. In recapitulation of embryonic events leading to Schwann cell maturation, we hypothesize that the Schwann cell-like cells acquire the switch to fate commitment through contact-dependant cues from incipient neurons of the developing dorsal root ganglia. To address this, Schwann cell-like cells derived from adult rat bone marrow were cocultured with neurons purified from embryonic dorsal root ganglia. A cell-intrinsic switch to the Schwann cell fate was achieved consistently and the cell progeny maintained expression of the markers S100β, p75^NTR , GFAP, P0 and Sox 10 even without exogenous differentiation-inducing factors or neurons. In vitro formation of MBP-positive segments under myelinating conditions by the cell progeny was comparable to that by sciatic nerve-derived Schwann cells. Controls in which Schwann cell-like cells were barred from direct contact with neurons in coculture reverted to SMA/CD90-expressing myofibroblasts. We demonstrate therefore for the first time fate commitment among bone marrow-derived Schwann cells. The therapeutic potential of these cells may be tested in future transplantation studies. (206 words)     

Beneficial effects of BV2 cell on proliferation and neuron-differentiating of mesenchymal stem cells in the circumstance of injured PC12 cell supernatant

Objective The microglias is the representative of immune cells in the brain. It plays dual roles of both repairing and damaging in injured nervous system, and works as an inevitable component of the circumstance of injured neurons. This study was aiming at the effects of the microglias on the biological activities of mesenchymal stem cells （MSCs） in the circumstance of injured neurons. Methods MSCs were obtained by primary culture. We adopted PC12 cells （PC12） and BV2 cells （BV2） to substitute for neurons and microglias, respectively. PC12 were injured by aged Aβ1-40 and the supernatant of the injured PC12 was used to set up the circumstance of injured neurons. Transwells were used for co-culture of BV2 and MSCs, which allowed the independent detection of cells after co-culture. Immunofluorescence was used to identify MSCs and neuron-differentiating cells with CD44 and neuron specific enolase （NSE） staining, respectively. MTT assay was adopted to measure the proliferation. Results In the circumstance of both BV2 presence and injured PC 12 supernatant incubation, either the proliferation or the differentiation of MSCs reached the highest, which seemed to be contradictory, but we gave our explanations. With the BV2 co-culture, the proliferation of MSCs tend to be higher, but the neuron-differentiating MSCs were similar to those incubated without BV2 co-culture either in normal or injured in PC12 supernatant. With the incubation of injured PC12 supernatant, the neuron-differentiating cells were significantly higher than that of control （P 〈 0.05）. Conclusion In the circumstance of injured neurons, microlgias tend to promote the MSCs proliferation. Although not helpful in neuron-differentiating, microglias did not exert any negative effect either.     

Morphological characteristics of p75 neurotrophin receptor-positive cells define a new type of glial cell in the rat dorsal root ganglia

In the dorsal root ganglia (DRG), two types of glial cells (Schwann cells and satellite glial cells) have been identified based on cell morphology and expression of specific markers. In the present study, we observed unknown glial cells that were positive for p75 neurotrophin receptor (p75NTR), and therefore were immunohistochemically and ultrastructurally characterized for the first time. These cells exhibited stronger immunoreactivity against an anti-p75NTR antibody than the DRG neurons (hereafter referred to as p75NTR++ cells). Moreover, these cells covered the glial cells surrounding proximal process of the large-diameter DRG neurons. The proximal process is called “dendro-axon.” The p75NTR++ cells were predominantly distributed where the first myelinating Schwann cells appear. The p75NTR++ cells were also positive for the pan-glial cell markers S100, nestin, and Sox10, but negative for fibroblast and macrophage markers. Moreover, they were negative for a satellite glial cell marker, inwardly rectifying potassium channel Kir4.1, as well as a nonmyelinating Schwann cell marker, glial fibrillary acidic protein. In addition, their morphological features were distinct from those of the myelinating Schwann cells. To investigate the three-dimensional ultrastructure of the p75NTR++ cells, we used array tomography combined with correlative light and electron microscopic observation. Three-dimensional ultrastructural observation revealed that the p75NTR++ cells loosely covered glial cells around the dendro-axons with highly ramified processes. Glial cells with these morphological features have not been reported before, indicating that the p75NTR++ glial cells are a new glial cell type in the DRG. Our results will give new insights into cell–cell relationships.     

Metabolic Control of Sensory Neuron Survival by the p75 Neurotrophin Receptor in Schwann Cells

We report that the neurotrophin receptor p75 contributes to sensory neuron survival through the regulation of cholesterol metabolism in Schwann cells. Selective deletion of p75 in mouse Schwann cells of either sex resulted in a 30% loss of dorsal root ganglia (DRG) neurons and diminished thermal sensitivity. P75 regulates Schwann cell cholesterol biosynthesis in response to BDNF, forming a co-receptor complex with ErbB2 and activating ErbB2-mediated stimulation of sterol regulatory element binding protein 2 (SREBP2), a master regulator of cholesterol synthesis. Schwann cells lacking p75 exhibited decreased activation of SREBP2 and a reduction in 7-dehydrocholesterol (7-DHC) reductase (DHCR7) expression, resulting in accumulation of the neurotoxic intermediate, 7-dehyrocholesterol in the sciatic nerve. Restoration of DHCR7 in p75 null Schwann cells in mice significantly attenuated DRG neuron loss. Together, these results reveal a mechanism by which the disruption of lipid metabolism in glial cells negatively influences sensory neuron survival, which has implications for a wide range of peripheral neuropathies.SIGNIFICANCE STATEMENT Although expressed in Schwann cells, the role of p75 in myelination has remained unresolved in part because of its dual expression in sensory neurons that Schwann cells myelinate. When p75 was deleted selectively among Schwann cells, myelination was minimally affected, while sensory neuron survival was reduced by 30%. The phenotype is mainly due to dysregulation of cholesterol biosynthesis in p75-deficient Schwann cells, leading to an accumulation of neurotoxic cholesterol precursor, 7-dehydrocholesterol (7-DHC). Mechanism-wise, we discovered that in response to BDNF, p75 recruits and activates ErbB2 independently of ErbB3, thereby stimulating the master regulator, sterol regulatory element binding protein 2 (SREBP2). These results together highlight a novel role of p75 in Schwann cells in regulating DRG neuron survival by orchestrating proper cholesterol metabolism.     

Adipose-derived stem cells differentiate into a Schwann cell phenotype and promote neurite outgrowth in vitro

Experimentally, peripheral nerve repair can be enhanced by Schwann cell transplantation but the clinical application is limited by donor site morbidity and the inability to generate a sufficient number of cells quickly. We have investigated whether adult stem cells, isolated from adipose tissue, can be differentiated into functional Schwann cells. Rat visceral fat was enzymatically digested to yield rapidly proliferating fibroblast-like cells, a proportion of which expressed the mesenchymal stem cell marker, stro-1, and nestin, a neural progenitor protein. Cells treated with a mixture of glial growth factors (GGF-2, bFGF, PDGF and forskolin) adopted a spindle-like morphology similar to Schwann cells. Immunocytochemical staining and western blotting indicated that the treated cells expressed the glial markers, GFAP, S100 and p75, indicative of differentiation. When co-cultured with NG108-15 motor neuron-like cells, the differentiated stem cells enhanced the number of NG108-15 cells expressing neurites, the number of neurites per cell and the mean length of the longest neurite extended. Schwann cells evoked a similar response whilst undifferentiated stem cells had no effect. These results indicate adipose tissue contains a pool of regenerative stem cells which can be differentiated to a Schwann cell phenotype and may be of benefit for treatment of peripheral nerve injuries.     

脊髓来源神经干细胞分离培养及向雪旺氏细胞的诱导分化

目的 探讨新生大鼠脊髓来源神经干细胞(NSCs)的分离培养及在体外一定条件下向周围神经雪旺氏细胞分化的可行性. 方法 分离新生大鼠的脊髓组织,在含有B27(终浓度1%)、碱性成纤维细胞生长因子(bFGF)和表皮生长因子(EGF)(终浓度均为20 μg/L)培养基中分离培养出NSCs.用复合诱导因子(10%FBS+5 μmol/L血小板凝集抑制剂+10 ng/mL bFGF+5 ng/mE血小板源性生长因子)在体外诱导NSCs分化为雪旺氏细胞.免疫荧光细胞化学方法[一抗为p75、S-100、神经胶质纤维酸性蛋白(GFAP)]鉴定体外诱导分化结果.结果 培养的新生大鼠脊髓组织细胞nestin染色表达阳性;分离培养的大鼠脊髓来源NSCs经诱导分化后形态类似雪旺氏细胞,免疫荧光细胞化学方法显示诱导后细胞表达雪旺氏细胞的表面标志,GFAP、S-100和P75表达阳性.结论 新生大鼠脊髓来源NSCs可以在体外诱导分化为雪旺氏细胞.