不同促细胞分裂因子对人神经干细胞定向分化的影响

目的观察不同促细胞分裂因子对人神经干细胞(NSCs)增殖及定向分化的影响。方法对人NSCs用无血清DMEM培养基行原代培养的同时,分别加入表皮生长因子(EGF)、碱性成纤维生长因子(bFGF)、神经生长因子(NGF)、维甲酸(RA)等因子,观察其对NSCs定向分化的作用。结果EGF培养的NSCs,克隆球形成慢且较松散,经血清诱导分化后,主要为星形胶质细胞,仅有少数神经元。而bFGF培养的NSCs则生长良好。在加血清诱导分化后,不同浓度bFGF培养的NSCs分化的细胞不同。bFGF与EGF共同培养的NSCs,其生长及神经球形成良好。在加血清诱导分化后,分化的神经细胞比例更接近脑内神经细胞的组分。NGF对神经球的形成无明显影响,但可促使其向神经元分化。RA使神经克隆球形成,可使NSCs直接分化为神经元细胞的比例明显增加。结论不同促细胞分裂因子对人NSCs的增殖及定向分化均有一定影响。     

人胚脑与脊髓神经干细胞体外生物学特性的差异

目的:探讨人胚脑源性神经干细胞和脊髓源性神经干细胞的体外培养和分化的差异。方法:从人胚脑组织和脊髓组织中分离培养神经干细胞,分为EGF组、bFGF组、EGF±bFGF组,在连续传代过程中观察并比较神经干细胞体外培养特性的差异:用血清诱导神经干细胞分化,观察其分化状况的不同。结果:从人胚脑组织分离的细胞在bFGF 单独存在时无法形成神经球,在EGF或EGF±bFGF存在时形成大量具有连续增殖能力的神经球;从人胚脊髓组织分离的细胞在EGF单独存在时无法形成神经球,在bFGF单独存在时只形成少量神经球,在EGF±bFGF存在时形成大量具有连续增殖能力的神经球。同样在EGF±bFGF存在的情况下,脑源性于细胞的增殖速度较快。经血清诱导后,脑组织来源的干细胞分化为NSE阳性细胞数明显多于脊髓组织来源的干细胞,二者之间的差异具有显著性(P<0．05)。结论:脑源性和脊髓源性神经干细胞在生长和分化方面有明显差别:脑源性神经干细胞可在bFGF或EGF士bFGF存在的情况下长期传代,而脊髓源性神经干细胞只能在EGF±bFGF存在的情况下长期传代,脑源性干细胞的增殖能力明显高于脊髓源性干细胞;脑源性干细胞较脊髓源性干细胞更易分化为神经元。     

人小龄胚胎神经干细胞的分离培养、扩增及鉴定

目的 探索人胚胎神经干细胞的体外分离培养条件，从而在体外大量扩增神经干细胞；并观察神经干细胞增殖、分化的特点。方法 从人胚胎脑分离神经干细胞，部分细胞冻存，另一部分细胞进行体外培养。采用无血清培养液，加入表皮生长因子(EGF)和碱性成纤维细胞生长因子(bFGF)刺激细胞增殖，进行体外扩增、传代培养。采用免疫荧光法鉴定神经干细胞和分化的神经元及胶质细胞。结果 从人胚胎脑分离的细胞在含有EGF和bFGF的无血清培养液中能形成大量的神经干细胞球，这些神经干细胞球可在体外扩增及传代培养。免疫荧光法鉴定神经干细胞球中大部分为神经上皮干细胞蛋白(nestin)表达阳性细胞。贴壁后可以分化出神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)表达阳性的细胞。经冻存后的胎脑细胞也能培养出具有同样特征的神经干细胞。结论 在含有EGF和bFGF的无血清培养液中，从人胚胎脑能分离培养出神经干细胞，并能在体外大量扩增。这为人类神经干细胞的进一步研究和应用提供了材料。     

Effect of growth factors on proliferation and phenotypic differentiation of human fetal neural stem cells

Human fetal neural stem cells (hNSCs) can be expanded in vitro by mitogens or growth factors, such as basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and/or leukemia inhibitory factor (LIF). Their effects on proliferation rate and differentiation pattern of hNSCs, however, have not been fully characterized. In this study, we cultured hNSCs in seven regimens, including bFGF, EGF, and LIF, either alone or in combinations. Cells were maintained as neurospheres in treatment media for various periods, up to six passages. A combination of bFGF, EGF, and LIF expanded hNSCs more efficiently than any other treatment as determined by counting total cell numbers using a trypan blue exclusion assay, a WST-1 cell viability assay, and a bromodeoxyuridine incorporation flow cytometric analysis. Differentiation patterns of hNSCs expanded under different conditions were also analyzed. We reported previously that hNSCs primed in vitro with a combination of bFGF, heparin, and laminin (FHL) induced neuronal differentiation toward a cholinergic phenotype. In this study, we show that the FHL priming increases neuronal differentiation while decreasing astroglial generation in all treatment groups as determined by immunostaining. However, cells proliferated under different growth factor conditions do vary in their phenotypic differentiation patterns. Particularly, significant generation of cholinergic cells was observed only in hNSCs expanded with EGF/bFGF or EGF/bFGF/LIF, but not with other treatment regimens, even when they are exposed to the same priming procedure. Our results indicate that hNSCs are highly plastic, with their proliferation and differentiation potential dependent on different growth factor treatments.     

Retinal neurospheres prepared as tissue for transplantation

The present work was conducted to study the cellular composition and developmental capacity of retinal neurospheres. Furthermore, the ability of grafted neurospheres to integrate into adult retinal tissue was studied in an in vitro model. Retinal progenitor cells isolated from rat embryos were expanded into neurospheres in vitro in the presence of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) and leukemia inhibitory factor (LIF). Neurospheres labeled with a lipophilic dye were placed onto explants, and tissue interactions were analyzed after 2-6 days of culture. Immunocytochemical analysis of neurospheres revealed the presence of neuronal and glial cells. Proliferating neuronal and glial cells were observed after 2 weeks, whereas the neuronal cell proliferation declined considerably after 4 weeks. Few apoptotic cells were observed in the neurospheres. Neurospheres cultured on explanted adult retina engrafted with the surrounding tissue, but progenitor cell migration into the explants was low. However, the grafted neurospheres appeared to limit the experimentally induced photoreceptor apoptosis in the surrounding explant tissue.     

成年骨髓间质干细胞体外诱导分化成神经细胞研究

目的：探索成年骨髓间质干细胞（ABMMSC）诱导分化为神经细胞（神经元和神经胶质细胞）的可行性，为ABMMSC在神经科学领域内的应用提供 参考。方法：以成年犬ABMMSC为实验对象，利用碱性成纤维细胞生长因子（bFGF）、表皮生长因子（FGF）、维甲酸（RA）、脑源性神经营养因子（BDNF）、胶质细胞系源性神经营养因子（GDNF）等作为增殖及分化诱导因子，采用两步法进行增殖培养，分化诱导；免疫细胞化学法进行细胞性质鉴定。结果：加入bFGF、EGF后增殖培养48h，换液、去除非粘附细胞，再增殖培养72h ,可见细胞分裂相（成纤维细胞样细胞）和簇样克隆形成（中小型细胞）。加入RA、BDNF、GDNF诱导3d，部分细胞有神经元特异性烯醇酶（NSE）、胶质纤维酸性蛋白（GFAP）成分表达；第10d可见有神经元、神经胶质形态样细胞形成。经细胞成分（NSE、GFAP）鉴定证实为神经元、神经胶质细胞。结论：ABMSC在体外培养条件下，经过bFGF、EGF、RA、BDNF、GDNF等因子的“程序性”作用，可以向神经元、神经胶质前体细胞及其终末细胞方向分化。     

人胚脊髓神经干细胞的分离培养和鉴定

目的:探讨人胚脊髓神经干细胞的体外培养和分化的方法,观察其增殖和分化特点。方法:利用无血清培养和单细胞克隆技术从人胚脊髓组织中分离培养出神经干细胞并用血清诱导其分化,应用免疫荧光细胞化学技术对培养细胞及其分化细胞进行鉴定。结果:从人胚脊髓组织分离的细胞在EGF单独存在时无法形成神经球,在bFGF单独存在时只形成少量神经球,在EGF和bFGF共同存在时形成大量具有连续增殖能力的神经球,表达神经干细胞的标志物Nestin,经血清诱导后分化为神经元、星形胶质细胞和少突胶质细胞并表达特异性抗原NSE、GFAP和CNP。结论:在体外培养条件下可从人胚脊髓组织中培养出神经干细胞,它可为神经干细胞的基础研究和临床应用提供材料。     

Developmental changes in progenitor cell responsiveness to cytokines.

Multipotent progenitor cells have been identified within periventricular generative zones of the developing and adult brain. To determine whether the environmental responsiveness of these cells changes during development, progenitor cells were cultured from embryonic, postnatal, and adult rat brain in the presence of either basic fibroblast growth factor (bFGF) or epidermal growth factor (EGF). Embryonic cells cultured as intact progenitor neurospheres proliferated more robustly in response to bFGF than to EGF, whereas proliferation of postnatal and adult progenitor cells was enhanced more by EGF than bFGF. Progenitor cells generated in the presence of either bFGF or EGF had the capacity to generate neurons, astrocytes, and oligodendrocytes at all developmental stages. Most embryonic and neonatal bFGF-generated cells differentiated predominantly into neurons, whereas late stage embryonic and neonatal EGF-generated progenitors largely remained in an undifferentiated state. However, later postnatal and adult progenitor species, irrespective of whether they were generated in the presence of bFGF or EGF, gave rise preferentially to astrocytes. Treatment with bone morphogenetic protein (BMP)2 or BMP7 enhanced astroglial differentiation and suppressed oligodendroglial differentiation of both EGF- and bFGF-generated progenitor species, suggesting that the effects of the BMPs are not dependent on EGF receptor activation. Thus, while central nervous system (CNS) progenitor cells retain multipotent capacity and responsiveness to the BMPs throughout development, they exhibit significant changes in other cellular response properties, perhaps reflecting differences in the requirements for specific generative versus regenerative events.     

人类神经干细胞的长期培养和传代

目的 探讨人类神经干细胞的体外培养条件及其传代的方法。方法 采用机械方法从胎脑中分离神经细胞，应用N2培养基进行培养，bFGF和EGF刺激细胞扩增；传统方法和对神经球切割的方法进行传代培养；应用免疫组织化学染色对培养的细胞及其分化的细胞进行鉴定。结果 从胎脑当中成功培养出人类的神经干细胞，培养条件下呈悬浮状态生长，形成神经球，绝大多数的细胞表达波形蛋白和Musashil两种神经干细胞的标志物；这种细胞可分化为神经元和星型胶质细胞，早期的培养有少量的少突胶质细胞；在这种培养条件下，神经干细胞生长速度较慢，而采用切割神经球的方法保持了细胞间的，神经干细胞可获得较大的扩增速度。结论 在体外的培养条件下，可从胎脑组织中培养出神经干细胞，它可做为中枢神经系统疾病移植治疗的潜在细胞来源。     

胚胎大鼠嗅神经干细胞的培养及分化特性

目的建立胚胎大鼠嗅神经干细胞(NSCs)体外培养方法,研究其增殖和分化特性.方法采用添加丝裂原的无血清培养基分离、培养胚胎14 d(E14)大鼠嗅球NSCs,应用免疫细胞化学方法鉴定培养的NSCs及自然分化为特异性神经细胞的类型,测定NSCs的生长曲线.结果从E14大鼠嗅球分离、培养出表达nestin,并能分化为神经元、星形胶质细胞和少突胶质细胞的NSCs.嗅NSCs的增殖依赖表皮生长因子(EGF)和碱性成纤维细胞生长因子(bFGF),其中EGF的促分裂增殖作用明显优于bFGF.结论从E14大鼠嗅球培养出具有自我增殖和多向分化潜能的NSCs.     

Epidermal growth factor (EGF), transforming growth factor-α (TGF-α), and basic fibroblast growth factor (bFGF) differentially influence neural precursor cells of mouse embryonic mesencephalon

Growth factors are key elements in the process of neural cell differentiation. We examined the effects of classical mitogens on neural precursor cells, by culturing mouse cells of the embryonic (13.5 days postcoitum) mesencephalon and treating them with epidermal growth factor (EGF), transforming growth factor-β (TGF-β), basic fibroblast growth factor (bFGF), nerve growth factor (NGF), and transforming growth factor-β (TGF- β). Our initial results show that EGF, TGF-α or bFGF, but not NGF or TGF-β, induced general proliferation of the cultured cells, followed by formation of colonies. Combinations of these three growth factors suggest that most cells with the capacity to form colonies responded to EGF, TGF-α, or bFGF. The number of colonies increased significantly when EGF, but not TGF-α, was used in combination with bFGF. Furthermore, a population responding only to EGF + bFGF was detected in the dorsal mesencephalon. The colony-forming activity of bFGF was dependent on insulin, but bFGF and insulin cooperation was indirect since we could not observe colony formation in subcultures of cells derived from colonies, even in the presence of insulin. Cells obtained from our colonies displayed neuronal and glial morphology and expressed markers of both neurons and astrocytes; nestin, a marker of neural precursor cells was also expressed in the majority of colonies. Growth factors also influenced neuronal maturation; the best neurite outgrowth was obtained from cells derived from bFGF-induced colonies cultured in the presence of EGF + bFGF. These data indicate the existence of neural precursor cells in the embryonic mesencephalon that respond differentially to growth factors. © 1995 Wiley-Liss, Inc.     

多细胞因子诱导成年鼠骨髓基质细胞向神经细胞分化的实验研究

目的观察大鼠骨髓基质细胞(rBMSCs)的生长特点及诱导条件下分化成神经细胞的能力,并对其机制进行初步探讨。方法以密度梯度离心分离骨髓基质细胞,在神经干细胞培养液中培养,采用四唑盐(MTT)法观察在培养液中添加碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)对BMSCs增殖的影响;观察添加脑源性神经生长因子(BDNF)、神经生长因子(NGF)和维甲酸(RA)对rBMSCs的诱导分化情况;采用免疫组织化学法(ABC)检测诱导后的细胞表达神经元特异性烯醇化酶(NSE)、神经元核蛋白(NeuN)和胶质原性纤维酸性蛋白抗体(GFAP)等特异性标志物的情况;以流式细胞分选确定神经元的比例。结果bFGF和EGF能在体外促进rBMSCs增殖,BDNF、NGF和RA能诱导rBMSCs来源的神经干细胞(NSCs)表达NSE、GFAP等特异性标志物。结论EGF、bFGF、BDNF、NGF、RA及适宜的培养液可使rBMSCs定向转化为NSCs,获得足够的目的细胞,进而分化为神经元样和神经胶质样细胞。     

Progenitor cells from the CA3 region of the embryonic day 19 rat hippocampus generate region-specific neuronal phenotypes in vitro

Progenitor cells that endure in different regions of the CNS after the initial neurogenesis can be expanded in culture and used as a source of donor tissue for grafting in neurodegenerative diseases. However, the proliferation and differentiation characteristics of residual neural progenitor cells from distinct regions of the CNS are mostly unknown. This study elucidated the characteristics of progenitor cells that endure in the CA3 region of the hippocampus after neurogenesis, by in vitro analyses of cells that are responsive to epidermal growth factor (EGF) or fibroblast growth factor-2 (FGF-2) in the embryonic day 19 (E19) rat hippocampus. Isolated cells from the E19 CA3 region formed neurospheres in the presence of either EGF or FGF-2, but the yield of neurospheres was greater with FGF-2 exposure, Differentiation cultures revealed a greater yield of neurons from FGF-2 neurospheres (60%) than from EGF neurospheres (35%). Exposure to brain-derived neurotrophic factor (BDNF) enhanced the yield of neurons from EGF neurospheres but had no consequence on FGF-2 neurospheres. A large number of neurons from EGF/FGF-2 neurospheres demonstrated clearly palpable morphological features of CA3 pyramidal neurons and lacked gamma-aminobutyric acid (GABA) expression. However, a fraction of neurons (17-20%) from EGF/FGF-2 neurospheres expressed GABA, and exposure to BDNF increased the number of GABAergic neurons (30%) from EGF neurospheres. Neurons from EGF/FGF-2 neurospheres also contained smaller populations of calbindin- and calretinin-positive interneuron-like cells. Thus, progenitor cells responsive to FGF-2 are prevalent in the CA3 region of the E19 rat hippocampus and give rise to a greater number of neurons than progenitor cells responsive to EGF. However, both FGF-2- and EGF-responsive progenitor cells from E19 CA3 region are capable of giving rise to CA3 field-specific phenotypic neurons. These results imply that progenitor cells that persist in the hippocampus after neurogenesis remain regionally restricted and hence retain their ability to give rise to region-specific phenotypic neurons even after isolation and expansion in vitro.     

Effects of Epimedium flavonoids on proliferation and differentiation of neural stem cells in vitro

Abstract

Objective: The purpose of this study is to investigate the effects of Epimedium flavonoids (EF), which is extracted from a traditional Chinese Epimedium herb, and its effect on the proliferation and differentiation of neural stem cells (NSCs) in vitro.

Methods: The single cells isolated from the hippocampi of 1 day old neonatal rats were cultured in a serum-free condition medium DMEM/F12 (1 : 1) with different concentrations of EF or 20 ng/ml epidermal growth factor (EGF) and 10 ng/ml basic fibroblast growth factor (bFGF). After 7 and 28 days, the neurospheres' diameters were measured. The formed neurospheres were cultured in the differentiation medium containing EF or 10% fetal bovine serum (FBS). After 12 hours and 7 days, immunofluorescent studies for nestin, Musashi-1, BrdU, β-III-tubulin, NF-200 and GFAP were performed. The number and lengths of 10–15 axons of NF-200 immunopositive cells were measured.

Results: The results showed that the isolated cells had the ability to propagate as neurospheres in the medium with 200 and 400 m g/ml EF, but without any EGF or bFGF, and the volume of neurospheres increase gradually from 7 to 28 days. In comparison with FBS control, the number of NF-200 positive neurons had significantly increased in the EF groups where the newborn neurons were morphologically more mature and able to migrate farther away from neurospheres than in the FBS control.

Discussion: The results demonstrate that EF effectively promotes the proliferation and differentiation of NSCs in vitro, suggesting that EF may have new properties of regulating central nervous system function by neurogenesis.     

底蜕膜间充质干细胞分化为多巴胺能样神经元

目的 探讨人胎盘底蜕膜间充质干细胞体外向多巴胺能样神经元分化的潜能,并优化诱导方案.方法 体外分离培养底蜕膜间充质干细胞,用表皮生长因子(EGF)+人碱性成纤维细胞生长因子(bFGF)+ B27添加剂和人音猬因子(SHH)+成纤维细胞生长因子8(FGF8)+forskolin+脑源性神经营养因子(BDNF)分两个阶段对其进行诱导;免疫细胞化学先后检测干细胞标记nestin和CD133、成熟神经元标记神经元特异性烯醇化酶(NSE)、神经胶质细胞标记胶质原纤维酸性蛋白(GFAP)、多巴胺能神经元标记酪氨酸羟化酶(TH)的表达;Western blot验证诱导后TH蛋白的表达;高效液相色谱-电化学检测诱导前后多巴胺的分泌.结果 经第一阶段诱导后,细胞形成漂浮生长的神经球,nestin和CD133均呈阳性表达;第二阶段诱导后,出现明显的神经元样形态,NSE、GFAP和TH均阳性表达,Western blot也显示TH蛋白的表达,多巴胺分泌量相比诱导前明显增加(P＜0.001).结论 底蜕膜间充质干细胞体外可分化为多巴胺能样神经元,可能成为帕金森病干细胞移植治疗新的种子细胞来源.     

碱性成纤维生长因子和表皮生长因子对脊髓神经干细胞增殖与分化的影响

目的观察碱性成纤维生长因子(bFGF)和表皮生长因子(EGF)对胚胎脊髓神经干细胞(NSC)增殖与分化的影响。方法从14 d胚胎大鼠的脊髓组织中分离培养脊髓NSC,并随机分为3组:EGF组、bFGF组和bFGF+EGF组。通过光镜观察不同时间点各组脊髓NSC克隆细胞团数量及直径大小,并采用免疫荧光染色检测各组脊髓NSC向神经元和星形胶质细胞分化的情况。结果①EGF组培养1、3、7 d后NSC克隆细胞团数量和直径均少于bFGF和bFGF+EGF组,差异有统计学意义(P0.05)。而bFGF+EGF组仅在培养1 d时克隆细胞团数量多于bFGF组,在培养3、7 d时差异无统计学意义。②EGF组分化细胞中神经元比例显著少于bFGF和bFGF+EGF组,星形胶质细胞数量明显大于bFGF和bFGF+EGF组,差异有统计学意义(P0.05)。而bFGF和bFGF+EGF组组间差异无统计学意义。结论 EGF对脊髓NSC克隆形成有一定作用,而bFGF能较好地促进克隆细胞团的形成及生长,两者联合应用在培养早期可显著促进克隆细胞团形成。EGF可诱导脊髓NSC更多分化为星形胶质细胞,而bFGF则可促进脊髓NSC向神经元分化。     

Establishment and characterization of a multipotential neural cell line that can conditionally generate neurons,astrocytes, and oligodendrocytes in vitro

In the mammalian central nervous system (CNS), multipotential neural stem cells in the neuroepithelium generate the three major types of neural cells, namely, neurons, astrocytes, and oligodendrocytes. To explore the molecular mechanisms underlying proliferation and differentiation of these neural stem cells, we established a cell line named MNS-57 from the embryonic day 12 rat neuroepithelium by introducing the mycer fusion gene, in which c-myc can be conditionally activated by adding oestrogen to the culture medium. MNS-57 cells expressed nestin, vimentin, and the RC1 antigen, which are potential markers for neural stem cells. We show that under particular culture conditions, MNS-57 cells can conditionally generate neurons, astrocytes, and oligodendrocytes in vitro, indicating that they are likely to originate from multipotential neural stem cells. Incubating MNS-57 cells with either oestrogen, which activates mycer, or growth factors such as basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) stimulated their growth, and the combination of oestrogen and bFGF (or EGF) had a synergistically stronger mitogenic effect than the single factors. Furthermore, both c-myc activation and bFGF appeared to be necessary for the differentiation of MNS-57 cells, and only when stimulated by both signals simultaneously, the cells committed to generating multiple neural cell types. Thus, the property of the cell line is unique in that its differentiation into neurons and glia can be conditionally manipulated invitro in an exogenous signal-dependent manner. We propose that the cell line described here will provide an useful in vitro model to understand genetic and environmental mechanisms that control the generation of neural cell diversity in the CNS. © 1995 Wiley-Liss, Inc.     

人胎脑神经球及其构成细胞的成熟表型标志

目的 观察体外培养人胎脑神经干细胞球的形成，并鉴别球内是否有成熟的神经细胞存在，证实神经球的异质性。方法用添加表皮生长因子(EGF，20ng／mL)和成纤维细胞生长因子2(FGF2，10ng／mL)的无血清N2培养基，从人胎脑培养获得神经干细胞球，在倒置显微镜下观察其形成过程，用间接免疫荧光技术检测神经球的细胞表型标志神经丝（NF)、胶质纤维酸性蛋白(GFAP)、半乳糖脑苷脂(GalC)和巢蛋白(nestin)的表达，并计算出阳性细胞球的百分比。结果前6d，培养中形成大量的细胞集落，检测的细胞标志中，只有nestin ，其余标志均为阴性。培养至12d左右，细胞球的数量减少约1／3．球体的形态和大小也不相同。培养30d的神经球中，69．2％神经球为NF ，81．8％为GFAP ，100％为nestin ．未观察到GalC 。结论神经球有个成熟过程，神经球之间具有异质性,球内的细胞构成也具有异质性。     

1101/神经干细胞超顺磁性氧化铁标记及对体外诱导分化的影响

目的 研究超顺磁性氧化铁（Superparam agnetic iron oxide）体外标记人神经干细胞的效果,为将来临床上应用磁共振成像（MRI）追踪标记细胞奠定基础。 方法 取孕13周人胚胎脑组织,制备成单细胞悬液,用添加表皮生长因子（EGF,20ng/ml）和碱性成纤维细胞生长因子（bFGF,20ng/ml）的体外培养人神经干细胞球,并做传代及诱导分化。利用Feridex标记分散成单细胞的神经干细胞,行普鲁士蓝染色检测标记阳性率,5%胎牛血清（FBS）诱导标记后的神经干细胞分化,并用免疫荧光染色检测分化细胞。 结果 培养一周即可见大量的神经干细胞球,Feridex标记神经干细胞行铁染色显示胞浆中含有铁颗粒,Feridex标记的神经干细胞阳性率达90%。标记后的神经干细胞经5% FBS诱导分化能够分化为神经元和星形胶质细胞 。 结论 Feridex 能够标记神经干细胞,标记不影响神经干细胞的生物学特性,标记后能够正常体外扩增培养和定向诱导分化,本研究为神经干细胞临床移植追踪提供了新的方法。