Lack of Teratogenicity of 3-Chloro-4-(Dichloromethyl)-5-Hydroxy-2 (5H)-Furanone (MX) in Embryonic Mouse Palate Culture in vitro

3-Chloro-4-(dichloromethyl)-5-hydroxy-2(5 H )-furanone (MX) is known as a by-product of wood pulp manufacture and a contaminant of chlorinated drinking water. Since our previous studies (Teramoto et al., 1998, 1999) demonstrated in a micromass in vitro test a strong inhibitory effect of MX on rat embryo cell differentiation, the potential teratogenicity was investigated in this study by using a suspension organ culture system. Twelve-day mouse embryo palatal explants were cultivated for 72 hr in the MX-containing medium at a concentration of 0, 1, 10, 100 or 300 μg/ml and examined for closure of the palatal shelves. All control explants showed almost complete closure of the palatal shelves. Similar results were also obtained in the MX-treated explants at concentrations up to and including 100 μg/ml. Immunohistochemistry revealed no difference between the control and MX-treated explants in distribution of PCNA-and TUNEL-positive cells in the palatal mesenchyme and medial edge epithelium, respectively. When the MX concentration was raised to 300 μg/ml, palatal shelves remained wide open. However, histopathology revealed extensive pyknosis of the mesenchymal cells and loss of the epithelium. These results may indicate that MX is cytotoxic against the mouse palate at a high concentration, and that it has no cleft-palate inducing effects in mice.     

应用5-杂氮脱氧胞苷诱导人羊水来源间充质干细胞向成肌细胞样细胞分化的体外研究

目的 探讨5-氮杂脱氧胞苷(5-AzaC)体外诱导人羊水来源间充质干细胞向成肌细胞样细胞分化的可行性.方法 B超引导下穿刺抽得孕中期羊水,体外分离、培养得到羊水来源间充质干细胞,传代后计数细胞并绘制生长曲线.体外成骨、成脂诱导观察人羊水来源间充质干细胞多向分化能力.采用5-AzaC对羊水来源间充质干细胞成肌诱导2周,观察细胞形态学变化,RT-PCR、免疫荧光染色方法鉴定成肌细胞特异mRNA及蛋白表达.结果 人羊水来源间充质干细胞体外培养后迅速进入对数生长期,培养7 d未达到平台期.茜素红和油红O染色证实人羊水来源间充质干细胞可诱导分化为成骨、成脂细胞样细胞.人羊水来源间充质干细胞经5-AzaC诱导2周逐渐变长梭形.而对照组细胞呈扁平多角形.免疫荧光染色及RT-PCR结果提示实验组细胞表达肌结蛋白(Desmin)、肌钙蛋白I(Tn I)、横纹肌辅肌动蛋白(α-Actinin)及mRNA;对照组呈阴性.结论 人羊水来源间充质干细胞体外增殖能力强,能分化为成骨、成脂细胞样细胞.5-AzaC能在体外诱导人羊水来源间充质干细胞向成肌细胞样细胞分化.     

In Vitro Developmental Toxicity of Aspirin and Its Major Metabolites on Cultured Fetal Mouse Palates

Palatal primordia of day-12.5 ICR mouse fetuses were cultured in a chemically-defined serumless medium by a suspension culture technique, and the developmental toxicity of aspirin and its metabolites on in vitro palatogenesis was studied. Explanted fetal palates were exposed in vitro for 72 hr to 0.5-2 mM aspirin (ASP), 0.25-2 mM salicylic acid (SA), 0.5-2 mM salicyluric acid (SUA), 1–2 mM 2,3-dihydroxybenzoic acid (3DHB), or 1–2 mM 2,5-dihydroxybenzoic acid (5DHB). After 72 hr culture, ASP at 2 mM and SA at 0.25 mM inhibited the growth and fusion of palatal shelves, and SUA at 1 mM prevented palatal fusion. On the other hand, 3DHB and 5DHB did not exert any significant toxic effects on cultured palates at concentrations up to 2 mM. Judging from the 50% inhibitory concentration (IC₅₀), SA (IC₅₀= 0.9 mM) was the most toxic of the 5 compounds tested, with a decreasing order of ASP (IC₅₀= 1.5 mM), SUA (IC₅₀= 1.6 mM), and DHBs (IC₅₀= over 2 mM for both 3DHB and 5DHB). With respect to developmental toxicity, cultured fetal mouse palates showed the susceptibility to aspirin and its metabolites which is intermediate between the susceptibility of rat embryos in vivo and that of postimplantation rat embryos cultured in vitro. The significance of fetal organ culture for evaluating developmental toxicity of chemicals is also discussed.     

Susceptibility of Day-12.5 and Day-13.5 Fetal Mouse Palates Cultured in Vitro to 5-Fluorouracil and Hydroxyurea

Abstract The maxillary regions of day-12.5 and day-13.5 ICR mouse fetuses were cultivated in a chemically-defined serumless medium by a suspension culture technique to examine the toxic effects of 5-fluorouracil (5-FU) and hydroxyurea (HU) on cultured palates and to compare the sensitivity of fetal mouse palates at different stages of development. The palates of day-12.5 and day-13.5 fetal mice were explanted and exposed in vitro for 72 hr to 0.1–50 μg 5-FU/ml or to 5–76 μg HU/ml. 5-FU inhibited the growth and fusion of day-12.5 palatal shelves in vitro dependently on its concentrations. Day-13.5 palates were significantly less sensitive to 5-FU than day-12.5 palates, and the minimal toxic concentrations (MTCs) of 5-FU were 0.1 and 10 μg/ml for day-12.5 and day-13.5 fetal palates, respectively. HU inhibited the in vitro growth and fusion of day-12.5 fetal palatal shelves in a concentration dependent manner, but only slightly suppressed the growth of day-13.5 fetal palates. The MTCs of HU were 19 and 76 μg/ml for day-12.5 and day-13.5 fetal palates, respectively. Therefore, day-12.5 fetal mouse palates (at stage-1 or earlier stages of palatogenesis) seemed significantly more susceptible to these teratogenic chemicals than day-13.5 fetal palates (at stages 2–3 of palatogenesis). The palates of day-12.5 ICR fetal mice may be more suitable than day-13.5 palates for in vitro teratogen screening and for the study of mechanisms of normal and abnormal palatogenesis.     

正常骨髓基质细胞对全反式维甲酸诱导分化HL-60细胞的作用

目的研究BMSC对ATRA抑制HL-60细胞增殖、诱导其分化的作用。方法分别将融合的基质细胞层(SC)、2%戊二醛固定的基质细胞层(FSC)和基质细胞,条件培养液(SCM)与HL-60细胞共同培养。SC、FSC、SCM组分别加或不加ATRA为实验组,另设阴性对照和ATRA对照组。通过细胞计数观察细胞增殖情况及通过细胞形态观察,NBT还原试验,分析细胞分化状态。结果细胞计数示SC、FSC、SCM各组均能抑制悬浮生长的HL-60细胞的增殖;SC和FSC能增强ATRA对HL-60细胞的抑制增殖作用(P<0·01)。SC、FSC、SCM各组的诱导分化率、NBT还原率等分化指标均与阴性对照组无差异(P>0·05);SC和FSC可促进ATRA对HL-60细胞的诱导分化作用(P<0·01)。结论BMSC可增强ATRA对悬浮生长的HL-60细胞的抑制增殖和诱导分化作用,这可能是通过BMSC与HL-60细胞之间的相互作用调节的。     

Enhanced adenosine 3':5' - monophosphate response to beta-adrenergic stimulation in cystic fibrosis fibroblasts after removal of conditioned medium

Skin fibroblasts derived from 6 patients with cystic fibrosis (CF), 1-6 months old, and from 6 age matched donors were investigated for their ability to accumulate cyclic adenosine 3':5'-monophosphate (c-AMP) in response to isoproterenol and prostaglandin E1 (PGE1) using strictly defined culture conditions. In order to obtain, as far as possible, constant protein content and cell number, cultures were synchronized in the early G1 phase of the cell cycle by growing them in serum free medium before adding stimulating drugs. There were no statistically significant differences both in basal c-AMP or after incubation with theophylline alone. When cultures were thoroughly washed prior to stimulation, c-AMP accumulation in response to isoproterenol was consistently higher (p less than 0.001) in CF than in normal fibroblasts, whereas response to PGE, did not differ significantly. This difference in response cannot be attributed to differences in dose- or time-response curves, or to differential escape of cAMP into the culture medium. Returning the conditioned media (CM) to the cultures after the washing procedure, or omitting the washing procedure altogether, normalized the cAMP response of CF cells. These data indicate that CF fibroblasts "delete" or "add to" the conditioned medium a substance when washed out of the cultures leave the cells hypersensitized to beta-adrenergic stimulation.     

Effects of antimicrobial agents used for therapy of CNS infections on dissociated brain cell cultures

The prediction, measurement, and monitoring of neurologic toxicity of antibacterial agents is an exceedingly difficult matter. In this study we investigated if in vitro exposure of cultured brain cells to antibacterial drugs could predict neurotoxicity in man. Effects of antibiotics used for therapy of bacterial CNS infections on growth and differentiation in dissociated rat brain cell cultures were studied over 24 days in culture, the drugs being added from 10 to 17 days in culture, the main differentiation phase of rat CNS cells. Our results demonstrated a reversible inhibition of cerebral sulfate transferase activity (p less than 0.001 or less than 0.01) and to a lesser extent (p less than 0.001 or NS) of DNA synthesis in brain cell cultures by the highest concentrations studied of amikacin, cefuroxime, and ceftazidime which correspond to peak cerebrospinal fluid values attained by intraventricular therapy in patients. Accumulation of DNA reflects brain cell growth whereas cerebral sulfate transferase activity parallels brain cell differentiation. Our findings indicate that intraventricular therapy could be more toxic with amikacin, cefuroxime, and ceftazidime than with penicillin, chloramphenicol, or ceftriaxone. Thus, this brain cell culture model might become a supplement, complement, or even alternative technique for neurotoxicity assessment of antibiotics with proven or potential value for therapy of CNS infections.     

Cerebrospinal fluid 309-1309-1309-1in neonates with central nervous system infections

Beta₂-microglobulin (₂m) determination in CSF of 72 neonates who underwent a spinal tap as part of a sepsis or meningo-encephalitis workup was performed to evaluate the usefulness of this test in the diagnosis of CNS infections. ₂m was measured by enzyme immunoassay. Sixty neonates had sterile culture and normal neurological status at discharge. Twelve infants had CNS infections: 8 bacterial meningitis, 3 TORCH infections (T=toxoplasmosis, O=others, R=rubella, C=cytomegalovirus and H=herpes simplex) and 1 viral meningitis. Neonates with CNS infection exhibited significantly higher CSF ₂m levels compared to neonates with sterile culture (6.24±2.66 vs 1.74±0.5 mg/l;P<0.0001). CSF ₂m levels did not correlate with the white cell count, total protein concentration or glucose level in CSF. When serum and CSF levels were measured simultaneously, the CSF ₂m level was significantly higher than the corresponding serum level in patients with CNS infection (6.98±2.5 vs 3.2±0.25 mg/l;P<0.01). Sensitivity, specificity, and predictive values were estimated for different cut-off points. The best operational diagnostic cut-off value was 2.25 mg/l. Receiver operating characteristic curve analysis showed an appropriate trade-off between specificity and sensitivity and indicated that CSF ₂m was accurate in distinguishing between neonates with and without CNS infection.Conclusion CSF ₂m may be a useful ancillary tool in neonates when CNS infection is suspected.Presented in part at the European Society for Paediatric Research Meeting, Edinburgh, September 1993     

Differentiation of neurospheres from the enteric nervous system

The enteric nervous system (ENS) derives from neural crest cells, which migrate from the neural tube into the developing gut. The neuronal and glial precursor cells migrate mainly from the oral towards the anal end of the gastrointestinal tract. So far, knowledge about the multipotent influences upon the ENS development, especially its neurotrophic support, derives mainly from knock-out models. The in vitro technique of isolating enteric neuronal precursor cells allows to study the effects of various factors upon their appropriate development in more detail. We therefore adapted the method of growing neurospheres, which are agglomerates of neuronal precursor cells and differentiated neurones and glial cells, from the central nervous system (CNS) for the ENS. The gut of NMRI mice at E12 were dissected, mildly dissociated and plated in 25-cm² culture flasks. The cultures were maintained in N1 supplemented DMEM/F12 medium with the appropriate neurotrophin cocktails (bFGF, GDNF, Neurturin, CNTF). After several days in culture most of the cells die, while the surviving cells form clusters from which domes, and later spheres arise. The spheres could be harvested and processed for further experiments. First investigations revealed, that the amount of precursor cells was much less in enteric neurospheres as seen in corresponding cultures from the CNS. We found about 43% HNK-1-NCAM+ in enteric and approximately 90% Nestin-+ cells in midbrain neurospheres. Differentiation studies of the enteric neurospheres showed that especially ciliary neurotrophic factor (CNTF) increased the number of enteric neurones (PGP positive), while the amount of HNK-1 precursor cells decreased under the influence of all tested neurotrophins but GDNF. The culture of the freshly dissociated enteric neurospheres in a three-dimensional matrix yielded a secondary network which allows to investigate the pattern formation of the ENS. The generation of enteric neurospheres and the following differentiation and 3D culture in vitro can increase our knowledge of the amount and time point of neurotrophic as well as the ECM-protein influence upon the appropriate development of the ENS.     

血管生成抑制剂TNP-470抑制血管瘤血管内皮细胞增殖的体外实验研究

目的 观察血管生成抑制剂TNP-470对血管瘤血管内皮细胞体外增殖的影响，并与氢化可的松相比较。方法 选择一草莓状血管瘤，采用M199培养液按组织块法进行血管内皮细胞(VEC)体外培养。传至第3代，VEC分为6组：组1、2、3在培养液中加入内皮细胞生长支持物(ECGS)，组1为对照组，组2加入氢化可的松，组3加入TNP-470；组4、5、6培养液中加入ECGS和雌二醇(E2)，组4为对照组，组5加入氢化可的松，组6加入TNP-470。培养后3、6、9d分别行细胞计数和3H-TdR掺入检测。结果 至第9d，组1VEC增殖明显，约增至3倍。组2、组3VEC增殖受到抑制，仅为组1的1/10(均P＜0．01)。组4VEC增殖最为显著，约为组1的2倍。组5、组6VEC增殖仍受到明显抑制，约为组4的1／7(均P＜0．01)。组2与组3、组5与组6VEC增殖状况相似(均P＞0．05)。结论 TNP-470能明显抑制ECGS或ECGS和E2联合诱导的血管瘤VEC体外增殖，抑制作用与氢化可的松程度相仿。     

重组人血管内皮抑素对鼠血管瘤内皮细胞株EOMA增殖的影响

目的 研究重组人血管内皮抑素(YH-16,恩度)对鼠源性血管瘤内皮细胞(EOMA)的抑制作用及其机制,并与血管瘤常用药物曲安缩松、平阳霉素(PYM)、干扰素α-2a相比较,观察单用及联合用药对EOMA细胞的抑制作用.方法 用MTT法检测不同浓度的YH-16对EOMA细胞作用24、48、72 h的抑制率,筛选出YH-16的最适抑制浓度.比较YH-16、曲安缩松、PYM、干扰素α2a四种药物单用以及YH-16、曲安缩松、平阳霉素两两联合作用对EOMA细胞的抑制率.采取流式细胞术测定YH-16作用EOMA细胞48h后,细胞凋亡率及Caspase-3的活性.结果 ①YH-16各浓度组在24h、48 h及72 h三个作用时段对EOMA细胞抑制作用显著(P＜0.01),并存在明显的剂量依赖关系;②选择200 gg/ml为YH-16的最适抑制浓度;③四种药物抑制作用由强到弱:PYM＞YH-16＞曲安缩松＞干扰素α-2a;④YH-16与曲安缩松或与PYM的联合效应均为拮抗效应,Q值均＜0.85;PYM与曲安缩松的联合效应为相加效应,0.85＜Q值＜1.15;⑤YH-16作用后细胞凋亡率随药物浓度增加而增高,Caspase-3表达在YH-16各浓度组均增强(P＜0.01).结论 YH-16对EOMA细胞有确切的抑制作用.该作用弱于PYM,而强于曲安缩松及干扰素α-2a,它主要是通过诱导细胞凋亡实现,且Caspase-3参与此过程.YH-16能拮抗曲安缩松及PYM抑制EOMA细胞增殖的作用,曲安缩松能增加PYM对EOMA的抑制增殖作用.     

Tissue culture of epithelial cells from urine. I. Serum-free growth of cells from newborn infants

The growth of epithelial cells from the urine of newborn infants was improved by the use of serum-free medium and a collagen type 1 matrix present on the growth surface of the culture vessel. The optimal concentrations and components of the serum-free medium consisted of a 1:1 mixture of Dulbecco's Modified Eagles' medium and Ham's F-12 medium supplemented with insulin (5 micrograms/ml), transferrin (5 micrograms/ml), selenium (5 ng/ml), and hydrocortisone (1 X 10(-7) M). The use of this medium allowed clonal isolates to undergo 25 generations and 5 passages with a doubling time of 24-36 hr with retention of original cell morphology. The culture of epithelial cells from the urine of newborn infants may provide a simple, reproducible system for the study of inborn errors of metabolism, especially those not expressed in fibroblast cultures.     

Intestinal mucosa of celiacs in remission is unable to abolish toxicity of gliadin peptides on in vitro developing fetal rat intestine and cultured atrophic celiac mucosa

Subfraction 2R of fraction 9 from a peptic-tryptic-pancreatic digest of wheat gliadin is known to be toxic in vivo to celiac patients. We have found that fractions 9 and 2R inhibit the in vitro development of fetal rat intestine and the increase of enterocyte height occurring in organ culture of atrophic celiac mucosa (0.1-0.5 mg/ml medium). Other peptide fractions of the gliadin digest are devoid of such in vitro effects. Subfraction 2R, after incubation with morphologically normal small intestinal mucosa of celiacs in remission and ultrafiltration, was still very active in both culture systems at low concentration (0.1 mg/ml); on the contrary, subfraction 2R was inactivated after incubation with normal mucosa. These results are compatible with the hypothesis that there is a mucosal defect in handling gliadin peptides in celiac disease, and suggest that there is either a primary (or secondary) enzyme deficiency or some other mechanism operating in the intestinal mucosa of celiac patients in remission.     

骨髓间充质干细胞条件培养基对缺氧大鼠神经元凋亡的影响

目的观察大鼠骨髓间充质于细胞（BMMSCs）条件培养基（B—CM））对缺氧大鼠大脑皮质神经元凋亡的影响。方法分离、培养、传代Wistar大鼠BMMSCs，细胞融合达90％时更换培养基，继续培养24h后收集培养上清液即B—CM；取培养8d的新生Wistar大鼠大脑皮质神经元，分正常对照组、缺氧组、B—CM组，分别于缺氧0、6h、复氧24h用四甲基偶氮唑盐微量反应比色（MTT）法检测各组细胞活性，用流式细胞仪和透射电镜技术检测神经元凋亡情况。结果神经元缺氧6h细胞活性较对照组明显降低，复氧24h后细胞凋亡率明显升高（P〈0．01）。B—CM培养后细胞活性和缺氧前无差异，细胞凋亡明显较少（P〈0．01）。结论B—CM对缺氧一复氧引起的神经元凋亡有明显的防护作用。     

Different effects of TrkA expression in neuroblastoma cell lines with or without MYCN amplification

BACKGROUND: Expression of the neurotrophin receptor TrkA is associated with a favorable prognosis in neuroblastoma (NB) and promotes growth inhibition and neuronal differentiation. Aggressive, MYCN-amplified NB tumors express little or no TrkA mRNA, suggesting that MYCN overexpression may inhibit TrkA expression. PROCEDURE: To study the interactions of TrkA expression and MYCN amplification in NB, we stably expressed the TrkA receptor in the MYCN single copy cell lines SH-SY5Y and NB69 as well as in the MYCN amplified cell lines CHP134 and IMR5. RESULTS: All four transfected cell lines demonstrated high TrkA expression and similar activation of the TrkA receptor and of mitogen-activated protein kinases as well as induction of immediate-early genes in response to nerve growth factor (NGF). Introduction of TrkA restored NGF responsiveness of SH-SY5Y and NB69 cells, as demonstrated by morphologic differentiation, growth inhibition, and enhanced survival in serum-free medium. However, no morphologic, growth, or survival responses to NGF were detected in MYCN-amplified CHP134 and IMR5 TrkA transfectants. CONCLUSIONS: Thus, transfection of TrkA into MYCN amplified NB cell lines only partly restored the TrkA/NGF signaling pathway, suggesting additional inhibitory effects of MYCN overexpression on TrkA signaling.     

High neuronal/astroglial differentiation plasticity of adult rat hippocampal neural stem/progenitor cells in response to the effects of embryonic and adult cerebrospinal fluids

Hippocampal neural stem/progenitor cells (hipp-NS/PCs) of the adult mammalian brain are important sources of neuronal and gial cell production. In this study, the main goal is to investigate the plasticity of these cells in neuronal/astroglial differentiations. To this end, the differentiation of the hipp-NS/PCs isolated from 3-month-old Wistar rats was investigated in response to the embryonic cerebrospinal fluid (E-CSF) including E13.5, E17-CSF and the adult cerebrospinal fluid (A-CSF), all extracted from rats. CSF samples were selected based on their effects on cell behavioral parameters. Primary cell culture was performed in the presence of either normal or high levels of KCL in a culture medium. High levels of KCL cause cell depolarization, and thus the activation of quiescent NSCs. Results from immunocytochemistry (ICC) and semi-quantitative RT-PCR (sRT-PCR) techniques showed that in E-CSF-treated groups, neuronal differentiation increased (E17>E13.5). In contrast, A-CSF decreased and increased neuronal and astroglial differentiations, respectively. Cell survivability and/or proliferation (S/P), evaluated by an MTT assay, increased by E13.5 CSF, but decreased by both E17 CSF and A-CSF. Based on the results, it is finally concluded that adult rat hippocampal proliferative cells are not restricted progenitors but rather show high plasticity in neuronal/astroglial differentiation according to the effects of CSF samples. In addition, using high concentrations of KCL in the primary cell culture led to an increase in the number of NSCs, which in turn resulted in the increase in neuronal or astroglial differentiations after CSF treatment.Key Words: Hippocampal neural stem/progenitor cells, Embryonic CSF, Adult CSF, Neuronal/astroglial differentiation     

PI3K抑制剂LY294002对鼠前脂肪细胞分化和C/EBPα及PPARγ表达的影响

目的：研究磷脂酰肌醇激酶（PI3K）抑制剂LY294002对小鼠前脂肪细胞分化和CCAAT增强子结合蛋白α（C/EBPα）及过氧化物酶体增殖物激活受体γ（PPARγ）表达的影响,探讨胰岛素受体底物（IRSs）/PI3K信号通路在前脂肪细胞分化中的作用。方法：小鼠3T3-L1细胞分为实验组和对照组,实验组加入LY294002（25 μmol/L）,对照组加入同体积DMSO。应用0.5 mmol/L 3-异丁基-1-甲基黄嘌呤(IBMX)、10-6 mol/L地塞米松和5 μg/mL胰岛素诱导两组前脂肪细胞分化,在培养的0 d、2 d、4 d、8 d分别收集细胞,实时PCR法及Western blot法检测细胞中C/EBPα和PPARγ表达水平。培养8 d油红O染色,观察细胞分化情况。结果：小鼠3T3-L1细胞基础状态下两组C/EBPα及PPARγ表达差异无统计学意义（P＞0.05）;诱导分化时实验组C/EBPα及PPARγ表达低于对照组（分别P＜0.05,P＜0.01）。油红O染色示实验组细胞无明显脂滴。结论：PI3K抑制剂LY294002能抑制小鼠前脂肪细胞分化及C/EBPα和PPARγ的表达,提示IRSs/PI3K信号通路可通过调节PPARγ和C/EBPα的表达在3T3-L1前脂肪细胞的分化中发挥重要作用。     

人胎脑神经干细胞体外培养及分化研究

目的研究人胚胎神经干细胞体外长期培养的条件、不同部位脑组织神经干细胞数量及其分化能力和特点。方法分离人胚胎海马、皮质、室周、纹状体脑组织，培养在含碱性成纤维细胞生长因子（b-FGF）、表皮生长因子（EGF）、B27、N2掭加剂的无血清培养基中，形成神经球，用有限稀释法进行克隆、传代。计算神经球数量，用溴脱氧尿苷（BrdU）标记神经球，使用免疫细胞化学法鉴定神经干细胞自主分化能力。结粜人胚胎海马、皮质、室周、纹状体部位脑组织均能培养出具有自我增殖能力的神经干细胞，其中海马所含神经干细胞最丰富。室周次之。BrdU检测有正在分裂、增殖的细胞。液氮冻存6个月的细胞复苏后仍具增殖分化能力。细胞贴壁分化后可形成Nestin、胶质原性纤维酸性蛋白（GFAP）、微管相关蛋白2（MAP2）表达阳性细胞。结论胚胎脑组织具有丰富的神经干细胞。具有自我更新和增殖能力，可长期培养。含b-FOF、EGF的无血清培养基能促进神经干细胞连续稳定增殖，保持神经干细胞特性。分离培养的神经干细胞可向神经元、胶质细胞分化。