改良Nash差速贴壁联合阿糖胞苷法体外分离培养大鼠嗅球及嗅黏膜源性嗅鞘细胞*☆

背景：目前常用的嗅鞘细胞培养方法有差速贴壁法、化学抑制法、抗体亲和吸附法、补体法等,各自均存在优缺点,单一使用某种方法时细胞纯化率较低。 目的：拟采用改良Nash差速贴壁+阿糖胞苷法体外分离纯化大鼠嗅球及嗅黏膜来源的嗅鞘细胞。 设计、时间及地点：细胞学体外对照观察,于2007-11/2008-05在武汉大学人民医院骨科实验室和消化内科实验室完成。 材料：10周龄Sprague-Dauley大鼠10只,由武汉大学人民医院实验动物中心提供,阿糖胞苷由武汉大学人民医院制备。 方法：完整取大鼠双侧嗅球及剪取鼻中隔后1/3嗅黏膜,剪碎后胰酶消化,制成单细胞悬液,按1×109 L-1密度接种于未包被的25 cm2玻璃培养瓶中,18~20 h后将细胞悬液转移至另一未包被的25 cm 2玻璃培养瓶中（第1次差速贴壁）,24 h后再将细胞悬液转移至经多聚右旋赖氨酸包被的25 cm2塑料培养瓶或经多聚右旋赖氨酸包被的6孔培养板中（第2次差速贴壁）,接种培养48 h后半量换液以去除杂细胞。在差速贴壁培养后二三天,加入3.0~5.0 pmol/L阿糖胞苷去除残余成纤维细胞。 主要观察指标：嗅鞘细胞的形态观察、分裂增殖情况、免疫荧光染色鉴定结果及纯度测定。 结果：体外培养24 h嗅球源性嗅鞘细胞即可贴壁,而嗅黏膜源性嗅鞘细胞多在培养四五天后贴壁,两种来源的嗅鞘细胞形态相似,以双极或梭形细胞为主,少量为3极及多突起形多级细胞,同时夹杂扁平、煎鸡蛋形细胞。纯化培养10 d的嗅鞘细胞,胶质纤维酸性蛋白、神经生长因子受体p75免疫荧光染色及神经生长因子受体p75+hoechs免疫荧光双染后大部分双极或多极细胞膜、胞体、突起呈阳性,细胞纯度可达90%以上。 结论：改良Nash差速贴壁+阿糖胞苷法可在体外成功分离培养出高纯度的嗅球及嗅黏膜源性嗅鞘细胞,嗅球源性嗅鞘细胞贴壁时间及分裂增殖程度优于嗅黏膜源性嗅鞘细胞。     

改良差速贴壁+无血清条件培养液纯化培养大鼠嗅鞘细胞*

背景：细胞移植是脊髓损伤的一种有效治疗手段,嗅鞘细胞被认为是最适合的种子细胞之一,但目前对其培养纯化的方法尚无公认标准。 目的：运用改良差速贴壁法+含同源嗅鞘上清液的无血清条件培养液来纯化培养嗅鞘细胞,以期建立一种新颖、经济、简单、实用的纯化培养成年大鼠嗅鞘细胞的方法。 设计、时间和地点：观察性对照实验。实验于2008-02/06在苏州大学干细胞与组织工程实验室完成。 材料：健康成年SD大鼠6只,体质量150~180 ｇ。 方法：①无菌条件取大鼠嗅球组织,消化、离心去除上清液后,用含体积分数为0.2胎牛血清的DMEM/F-12培养基重悬,稀释的单细胞混悬液均分成3份,分别用于单纯改良差速贴壁、单纯无血清条件培养液纯化和改良差速贴壁+无血清条件培养液纯化的实验。②单纯改良差速贴壁：将单细胞混悬液以8 000个/cm²密度种植于无包被的25 cm²培养瓶中。经12 h+ 24 h两次差速培养后,吸出细胞上悬液,计数板计算浓度后,按1×109 Ｌ-1浓度重新种植于载有多聚赖氨酸包被盖玻片的35 mm培养皿中继续培养3周。③单纯无血清条件培养液纯化：将单细胞混悬液用预先收集并制备的含同源嗅鞘培养上清液的无血清条件培养液直接种植于载有多聚赖氨酸包被盖玻片的35 mm培养皿中,孵育两三天后再换成含体积分数为0.1胎牛血清DMEM/F12培养基继续培养3周。④改良差速贴壁+无血清条件培养液纯化：经12 h +24 h两次差速纯化,吸出上悬液接种于培养皿中继续培养2.0~3.0 d后,改换成无血清条件培养液,孵育36~48 h后再换成含体积分数为0.1胎牛血清DMEM/F-12培养基继续培养。以后视情况每3~5 d半量换液1次,培养3周。 主要观察指标：运用倒置显微镜观察各组不同时间的嗅鞘细胞生长状况和形态学特征。采用GFAP、NGFRp75和Hoechst33258等抗体,于分离培养14 d后进行细胞免疫荧光染色并检测嗅鞘细胞的纯度。 结果：①倒置显微镜观察：差速后3 d内有大量细胞贴壁生长,细胞形态较混杂,辨认嗅鞘细胞较困难。差速后再运用无血清条件培养液2 d后,可见纯度较高的典型嗅鞘细胞形态,以双极梭形和多极为主,细胞突起细长。伴少量扁平状“油煎蛋样”细胞。继续培养至14 d可见细胞立体感及折光性最佳,数量和纯度最高。培养3周后3组细胞开始老化,活力明显下降,成纤维等杂细胞开始挤占原嗅鞘细胞生长空间。②免疫荧光染色显示嗅鞘细胞特异性表达GFAP和NGFRp75,单纯差速后嗅鞘细胞纯度仅有72%~75%,单纯无血清条件培养液纯化后仅为48%~52%,改良差速贴壁+无血清条件培养液纯化组纯度可达88%~92%。 结论：实验建立了完善的成年大鼠嗅鞘细胞培养纯化新方法。     

以差速贴壁与化学药物并胰酶限时消化法纯化新生大鼠嗅鞘细胞：优于单一差速贴壁和化学药物法吗？★

背景：目前国内外对嗅鞘细胞培养条件的报道各不相同，个别方法重复性较差，不利于实际应用。 目的：观察差速贴壁+化学药物+胰酶限时消化法纯化大鼠嗅鞘细胞的效果，并与化学药物法、差速贴壁法培养结果进行比较。 设计、时间及地点：细胞学体外对照观察，于2007-06/2008-06在福建医科大学人体解剖与组织胚胎学系实验室完成。 材料：新生2 d的SD大鼠8只，由福建医科大学试验动物中心提供。 方法：无菌条件下完整取出大鼠双侧嗅球，剥除嗅球表面的软脑膜及毛细血管网，剪成0.5 mm3的小块进行胰蛋白酶消化，过筛后按4.0×108 L-1密度种植于包被多聚左旋赖氨酸的培养瓶中进行原代培养。设立4组：①未经纯化的常规对照组。②化学药物组加入5 μmol/L阿糖胞苷抑制成纤维细胞分裂。③差速贴壁组采用Nash差速贴壁法纯化嗅鞘细胞。④差速贴壁+化学药物+胰酶限时消化组首先采用Nash差速贴壁法去除大部分成纤维细胞，而后对于少量残留的成纤维细胞加入阿糖胞苷处理，培养6 d贴壁细胞大部分融合后，加入1.25 g/L胰蛋白酶消化1 min，显微镜下见突起回缩、细胞变圆、少量细胞浮起时终止消化。 主要观察指标：嗅鞘细胞的形态，NGFR p75免疫细胞荧光染色结果。 结果：体外培养的新生大鼠嗅球嗅鞘细胞主要为双极或三极细胞，其突起细长；未经纯化的常规对照组培养7 d时视野内成纤维细胞已达60%以上，至14 d成纤维细胞完全长满；经过纯化处理的另外3组始终以嗅鞘细胞居多，嗅鞘细胞的形态与原代基本相同。存活的双极和3极嗅鞘细胞呈NGFRp75阳性反应，但化学药物组、差速贴壁组嗅鞘细胞纯度偏低，仅为75%；差速贴壁+化学药物+胰酶限时消化组嗅鞘细胞纯化效率明显增高，达85%以上。 结论：实验结果证实了差速贴壁+化学药物＋胰酶限时消化法的三合一操作技术，是一种高效率的纯化培养嗅球嗅鞘细胞的方法。     

阿糖胞苷结合神经生长因子体外纯化培养大鼠嗅鞘细胞*★

背景：由于嗅鞘细胞终生具有成髓鞘能力，如何利用简单而又经济的方法获得大量较高纯度的嗅鞘细胞是脊髓损伤研究的热点。 目的：分析阿糖胞苷结合神经生长因子对大鼠嗅球源性嗅鞘细胞纯化培养的可行性。 方法：将差速贴壁后的大鼠嗅鞘细胞首先用含10 mg/L阿糖胞苷的完全培养基培养24 h，然后用含体积分数为1%胎牛血清和25 μg/L神经生长因子的DMEM/F12培养基进行体外培养。观察嗅鞘细胞的生长变化，采用形态学和免疫组化染色对细胞及其纯度进行鉴定。 结果与结论：体外培养的大鼠嗅鞘细胞神经生长因子受体NGFRp75染色呈阳性反应，细胞呈双极和三级，伸出细长突出，并渐结成网状，在体外培养的第7天纯度为95%，第9天时为90%，且形态良好。结果提示阿糖胞苷结合神经生长因子是一种简单实用的嗅鞘细胞纯化培养方法。     

分离、培养及纯化嗅鞘细胞的简便方法

目的：建立一种免胰酶消化的剪切分散法提取嗅鞘细胞（olfactory ensheathing cells,OECs）并优化其纯化培养方法。方法：从成年SD大鼠解剖分离嗅球,显微镜下剥离软脑膜、剔除中央白质,将灰质成分置于含10%胎牛血清的DMEM/F12全培养基,眼科剪剪切至组织成糜状,吸管吹打15-20次, 混悬液免胰酶消化直接接种培养,每三天半量换液。原代培养至细胞大部分融合后,依次以低浓度胰酶（0.05%）有限消化法、30min短时差速贴壁法、抗有丝分裂法三步顺序纯化细胞,每步纯化结果作免疫荧光染色鉴定。结果: 原代培养中嗅鞘细胞快速贴壁成为优势细胞、生长旺盛,三步顺序纯化过程细胞纯度逐步提高,最终纯度可达95%以上并长时间维持。结论:剪切分散法提取OECs及其三步顺序法纯化培养经济实用、易重复,细胞培养周期短,能收获大量可控纯度的嗅鞘细胞供进一步移植实验及基础研究。     

用于移植的嗅鞘细胞的纯化培养方法研究

目的 探讨移植用成年大鼠嗅球嗅鞘细胞(OECs)的培养和纯化方法.方法 取成年SD大鼠嗅球,将分离消化所得细胞悬液进行接种.然后分别于接种后2 h、6 h和18 h 3个时间点进行单步骤差速贴壁法去除成纤维细胞,纯化嗅鞘细胞,纯化培养约11 d时采用神经生长因子受体P75(NGFRP75)和碘化丙啶(PI)双染OECs并对其纯度及细胞数量进行免疫荧光鉴定分析.结果 纯化培养11 d后在共聚焦显微镜下呈双染阳性的细胞为OECs,数量级达106/mL,形态以梭形、双极或三极突起为主,亦有少量多极或单极细胞.2 h、6 h和18 h组嗅鞘细胞纯度分别为(53±5)%、(73±8)%和(69±13)%,组间差异有统计学意义(F=11.766,P<0.001). 结论分别经2、6、18 h单步骤差速贴壁法纯化的OECs在纯度和数量级上均可满足移植用细胞的要求,且方法简单、经济、快捷.     

胰酶限时消化在嗅鞘细胞培养中的运用

目的探索出一套高效、实用的大鼠嗅球嗅鞘细胞(OECs)的培养和纯化方案。方法分离SD大鼠嗅球的外两层组织,剪切消化成细胞悬液进行接种。采用3种方法进行OECs的培养和纯化:(1)A组分别经6h、24h两次差速贴壁去除杂质细胞,培养至12d左右消化重悬细胞进行爬片分析。(2)B组分别经6h、24h两次差速贴壁去除杂质细胞,培养至12d左右经胰酶限时消化,留成纤维细胞于瓶壁,重悬的细胞进行爬片分析。(3)C组采用经典的Nash法(分别经18h、36h两次差速贴壁去除杂质细胞),培养至11d左右消化重悬细胞进行爬片分析。采用NGFRP75与碘化丙啶(PI)双染的方法进行OECs的鉴定和纯度分析。结果在共聚焦显微镜下呈双染阳性的细胞为OECs,OECs多数突起细长,呈双极或三极,少量呈单极或多极。A、B、C组所纯化的OECs纯度分别为(67.3±6.2)%、(83.7±7.7)%和(74.6±9.5)%,3组间比较有统计学差异(F=13.633,P<0.01),B组所得OECs纯度均较另两组高(P<0.05)。结论经6h、24h两次差速贴壁+胰酶限时消化可以获得较高纯度的OECs,能够满足动物实验的需要。     

牛垂体提取物、碱性成纤维细胞生长因子及Forskolin影响嗅鞘细胞增殖的量效关系

背景：生长因子参与调节嗅鞘细胞增殖、分化及代谢功能,因此有望通过应用其合理的序贯刺激形式,达到细胞数量地大量扩增。 目的：观察牛垂体提取物、碱性成纤维细胞生长因子及Forskolin对嗅鞘细胞增殖的影响及剂量-效应关系。 方法：采用原代培养技术,从新生大鼠的嗅球分离培养嗅鞘细胞,并用差速贴壁+化学药物+胰酶快速消化法纯化培养嗅鞘细胞,添加牛垂体提取物、碱性成纤维细胞生长因子及Forskolin 3种促增殖因子及其组合。MTT法检测各组嗅鞘细胞生长的情况。根据NGFRp75抗体的免疫细胞化学染色统计嗅鞘细胞的纯度和计算嗅鞘细胞的增殖率。 结果与结论：在嗅鞘细胞原代培养和纯化后,添加牛垂体提取物具有促进嗅鞘细胞增殖的效应,但未见典型的剂量-效应关系;添加碱性成纤维细胞生长因子具有促进嗅鞘细胞增殖的效应,且呈现典型的剂量-效应关系;添加Forskolin并不具有促进嗅鞘细胞增殖的效应。说明合理应用不同质量浓度的碱性成纤维细胞生长因子与Forskolin进行联合刺激,能有效促进细胞的增殖。结果表明序贯的使用合适浓度的生长因子组合,将是有效促进嗅鞘细胞增殖的合理策略,有助于解决脊髓损伤治疗中种子细胞来源不足这一问题。     

不同来源嗅鞘细胞移植治疗脑出血的比较

背景：嗅鞘细胞是目前已知用于移植细胞中惟一可以跨越周围神经系统与中枢神经系统边界的细胞。在众多的国内外文献中,大多以嗅球来源嗅鞘细胞作为观察对象;但采用嗅黏膜嗅鞘细胞移植具有来源方便、损伤小、可自体取材等优势。 目的：比较嗅球来源嗅鞘细胞和嗅黏膜嗅鞘细胞移植对脑出血后神经功能损伤恢复作用的差异。 设计、时间及地点：免疫组织化学水平的随机对照动物实验,于2005-10/2007-10在无锡市第三人民医院细胞实验室完成。 材料：选用健康雄性SD大鼠30只,按随机数字表法分为3组,对照组、嗅球来源嗅鞘细胞移植组及嗅黏膜嗅鞘细胞移植组各10只。 方法：分别获取人鼻中隔黏膜、人胚嗅球,进行嗅鞘细胞的分离培养纯化,取培养10 d的细胞用作细胞移植。取SD大鼠制备尾状核出血模型,嗅球来源嗅鞘细胞移植组及嗅黏膜嗅鞘细胞移植组各取10μL细胞悬液在立体定向下引导微量注射器向大鼠脑组织内匀速注射(1 μL/min),对照组注入等量培养基,注射点为右侧尾状核。 主要观察指标：观察两种来源嗅鞘细胞的体外培养情况。嗅鞘细胞移植后对动物行定期行为学评定,功能损伤越重,得分越高;并观察组织切片靶区域免疫细胞化学分析结果。 结果：从体外培养结果看,无论在形态上还是表型上,两者无明显区别。嗅球来源嗅鞘细胞移植组及嗅黏膜嗅鞘细胞移植组大鼠的Longa 5分制法及前肢放置试验评分在移植后第14,28,56天显著低于对照组（P < 0.05）,两移植组差异无显著性意义（P > 0.05）,移植组各时间段差异无显著性意义（P > 0.05）。 结论：用于移植修复神经功能的两种嗅鞘细胞在细胞特性、移植效果方面均无明显差异。     

改良成年大鼠嗅鞘细胞的原代培养

目的研究成年大鼠嗅鞘细胞(OECs)分离与培养的方法,为进一步进行基础研究以及临床应用提供高纯度的细胞.方法采用改良差速贴壁法从成年大鼠的嗅球培养出OECs,并用Forskolin和碱性成纤维细胞生长因子(bFGF)促增殖,最后进行OECs特异标记物NGFRp75、S-100的免疫细胞化学染色鉴定OECs.结果改良差速贴壁法成功培养出高纯度的OECs.结论此法简便、经济、纯化效率高,Forskolin和bFGF能够促进OECs的增殖.     

The olfactory mucosa,first actor of olfactory detection,is sensitive to glucocorticoid hormone

The olfactory mucosa (OM) is the primary site of odorant detection, and its axonal projections relay information to brain structures for signal processing. We have previously observed that olfactory function can be affected during a prolonged stress challenge in Wistar rats. The stress response is a neuroendocrine retro‐controlled loop allowing pleiotropic adaptive tissue alterations, which are partly mediated through the release of glucocorticoid hormones. We hypothesised that, as part of their wide‐ranging pleiotropic effects, glucocorticoids might affect the first step of olfactory detection. To study this, we used a number of approaches ranging from the molecular detection and functional characterisation of glucocorticoid receptors (GRs) in OM cells, to the study of GR acute activation in vivo at the molecular, electrophysiological and behavioural levels. In contrast to previous reports, where GR was reported to be exclusive in olfactory sensory neurones, we located functional GR expression mostly in olfactory ensheathing cells. Dexamethasone (2 mg/kg) was injected intraperitoneally to activate GR in vivo, and this led to functional odorant electrophysiological response (electro‐olfactogram) and OM gene expression changes. In a habituation/cross‐habituation test of olfactory sensitivity, we observed that DEX‐treated rats exhibited higher responsiveness to a complex odorant mixture. These findings support the idea that olfactory perception is altered in stressed animals, as glucocorticoids might enhance odour detection, starting at the first step of detection.     

Isolation, characterization, and genetic profiling of subpopulations of olfactory ensheathing cells from the olfactory bulb

Olfactory ensheathing cells (OECs) play a crucial role during neurogenesis of primary olfactory neurons. Transplantation of OECs is considered as a promising new therapy for central nervous system repair. Nevertheless, OECs are constituted of distinct subpopulations and their role during neurogenesis is not clearly understood. In particular, OECs from the olfactory bulb (OB) constitute a heterogeneous, but not yet isolated and characterized, population of cells. In our study, flow cytometry analyses of primary OB cultures, based on cell surface expression of low-affinity nerve growth factor receptor (p75), reveal the presence of two distinct populations of OECs. Indeed, some of them express a high level of p75 (P75High) and the other a low level of p75 (P75Low). Effects of OB microenvironment were assessed, and we were able to show that fibroblasts mediate the induction of these two populations through the secretion of soluble factors. To characterize P75High and P75Low OECs, cells were sorted based on their differential expression of p75. Microarray analyses revealed that P75High OECs overexpress genes implicated in modulation of extracellular matrix and cell sorting, whereas P75Low OECs overexpress genes involved in regulation of the inflammatory response and axonal guidance. These results permit, for the first time, to isolate the two distinct subpopulations of OECs from OB, and suggest their specific role during neurogenesis.     

Organization of projections from olfactory epithelium to olfactory bulb in the frog, Rana pipiens

One hypothesis for the coding of olfactory quality is that regions of the olfactory epithelium are differentially sensitive to particular odor qualities and that this regional sensitivity is conveyed to the olfactory bulb in a topographic manner by the olfactory nerve. A corollary to this hypothesis is that there is a sufficiently orderly connection between the epithelium and the olfactory bulb to convey this topographical coding. Thus we examined topography in the projection from epithelium to bulb in the frog, which has been the subject of numerous electrophysiological studies but has not yet been examined using modern neuroanatomical techniques. The tracer WGA-HRP was applied to the ventral or to the dorsal olfactory epithelium, or both. Anterograde transport of label to the olfactory bulb was seen after as few as 2 days; label was still present in the bulb as long as 21 days postinjection. In cases where WGA-HRP was applied to the entire epithelium, there was dense anterograde labelling of the ipsilateral olfactory bulb. In addition, a small medial portion of the contralateral bulb was labelled. Injections limited to either the ventral or dorsal epithelium produced patterns of anterograde labelling in the glomerular layer of the olfactory bulb, which varied with the size and location of the injection. With very large injections in either the dorsal or ventral epithelium, label appeared to be evenly distributed in the glomerular layer. With smaller injections in the ventral epithelium, there was heavier labelling in the lateral than in the medial portions of the glomerular layer, although light labelling was found in all regions of the glomerular layer. In contrast, injection sites restricted to the dorsal epithelium produced more anterograde labelling in the medial than lateral portions of the glomerular layer. These patterns extended throughout the dorsal-ventral extent of the bulb. Within the limits of the anterograde tracing technique used, we were unable to detect any systematic relationship between the pattern of labelling in the glomerular layer and the medial-lateral or rostral-caudal location of the injection site in either the ventral or dorsal epithelium. We conclude that in the frog, as in other amphibia, there is only a limited degree of topographic order between the epithelium and the olfactory bulb.     

Depth of olfactory sulcus and olfactory function

The aim of this study was to identify whether the depth of the olfactory sulcus relates to olfactory function in healthy subjects. Forty-four healthy, male volunteers (age range 22-45 years, mean age 28.3 years) were included in this study. Olfactory function was measured for phenyl ethyl alcohol odor thresholds, odor discrimination, and odor identification. Magnetic resonance imaging of the olfactory sulcus was performed immediately following olfactometry. Based on previous investigations the depth of the olfactory sulcus was measured in the plane of the posterior tangent through the eyeballs. Olfactory function correlated significantly with left-sided depth of the olfactory sulcus (r(44)=0.33, P=0.03); no such correlation was seen for the right side. In addition, olfactory sulcus depth was found to be significantly deeper on the right compared to the left side (t=5.61, P<0.001). The present results suggest that there is small, but significant relation between morphological brain structures and measures of olfactory function. Further, lateralization of olfactory sulcus depth may correlate to functional lateralization in the olfactory system. Thus, it may be carefully speculated that sensory input in the olfactory system results in cortical growth in the area of the olfactory sulcus, at least at some developmental stage.     

Visual, somatosensory, olfactory, and gustatory hallucinations.

Hallucinations that involve any of the sensory modalities may accompany a number of functional and organic conditions. Although characteristics of the hallucinations are not specific, they are characteristic and suggestive of specific disorders. Appropriate evaluation and treatment require consideration of the past psychiatric, neurologic, and medical history; assessment of accompanying psychiatric and neurologic signs and symptoms; and degree of response to conventional therapy. Any patient with hallucinations of recent onset or presenting a significant change in the nature of prior hallucinations, particularly when the patient does not respond to conventional therapy, deserves an evaluation to rule out treatable organic factors.     

Isolation, culture, and purification of olfactory mucosa-derived olfactory ensheathing cells using modified differential attachment with low concentration serum

BACKGROUND:Studies have demonstrated that olfactory mucosa can promote the regeneration and formation of axonal medullary sheath of injured neurons. To date, olfactory ensheathing cells (OECs) utilized in basic and clinical research arise primarily from the olfactory bulb mucosa. However, little is known regarding culture, purification, and biological properties of OECs . OBJECTIVE: To isolate and culture OECs utilized modified, differential attachment in combination with neurotrophic factor 3 (NT3) and low...     

Comparative gene expression profiling of olfactory ensheathing cells from olfactory bulb and olfactory mucosa

Olfactory ensheathing cells (OEC) have the ability to promote regeneration in the nervous system. Hence, they hold promise for cell therapy. Most of the experimental studies have investigated the role of OECs taken from olfactory bulb (OB). However, for a clinical human application, olfactory mucosa (OM) seems to be the only acceptable source for OECs. Many studies have compared the distinct ability of OECs from OB and OM to improve functional nerve regeneration after lesion of the nervous system. Nevertheless, the two populations of OECs may differ in several points, which might affect all fate after transplantation in vivo. We report here the first study which compares gene expression profiling between these two populations of OECs. It appears that OB‐OECs and OM‐OECs display distinct gene expression pattern, which suggest that they may be implicated in different physiological processes. Notably, OM‐OECs overexpress genes characteristic of wound healing and regulation of extra cellular matrix. In contrast, OB‐OECs gene profile suggests a prominent role in nervous system development. Hence, OB‐OECs and OM‐OECs fundamentally differ in their gene expression pattern, which may represent a crucial point for future clinical application. © 2010 Wiley‐Liss, Inc.     

Isolation, culture, and purification of olfactory mucosa-derived olfactory ensheathing cells using modified differential attachment with low concentration serum*☆

BACKGROUND：Studies have demonstrated that olfactory mucosa can promote the regeneration and formation of axonal medullary sheath of injured neurons. To date, olfactory ensheathing cells （OECs） utilized in basic and clinical research arise primarily from the olfactory bulb mucosa. However, little is known regarding culture, purification, and biological properties of OECs .
OBJECTIVE： To isolate and culture OECs utilized modified, differential attachment in combination with neurotrophic factor 3 （NT3） and low concentration serum to explore an optimal in vitro culture method for OECs.DESIGN, TIME AND SETTING： Single-sample observation was performed at the Medical Experimental Center of Stomatology College, Xi＇an Jiaotong University between March 2006 and December 2007. MATERIALS： Twelve samples from aborted embryos, 4-6 months, were used to isolate OECs; rabbit-anti-human p75NTR and glial fibrillary acidic protein （GFAP） antibody were provided by Sigma, USA. METHODS： The differential time was six hours. This was repeated twice, based on Nash＇s differential attachment. Attached OECs were cultured in DMEM-F12 culture medium containing 10% fetal bovine serum （FBS） or 2.5% FBS and NT3. MAIN OUTCOME MEASURES： OEC morphology was observed, and p75NTR and GFAP immunocyto-chemistry was used for identification and purity detection. RESULTS： Some cells attached after three days in culture. Several cells possessed short neurites with good refractivity. Some shuttle-shaped fibroblasts could be seen. On day six, more cells attached, exhibiting a three-dimensional appearance. Many cells appeared dipolar or tripolar, with slender neurites, and fibroblasts were clustered. On day nine, the number of dipolar or tripolar cell bodies with slender neurites was increased, and fibroblasts were clustered. On day 15, fibroblasts occupied the majority of the bottom of the culture bottle, with several OECs surviving at the upper layer. OECs were positive for P75NTR and GFAP expression, as identified by an im     

Deviant olfactory experiences,magical ideation,and olfactory sensitivity: a study with healthy German and Japanese subjects

Little is known about the relationship between olfactory hallucinations and olfactory sensitivity in psychiatric populations. However, in healthy subjects, a 'psychotic-like' feature, namely magical ideation, has been linked to deviant olfactory experiences. We thus assessed olfactory sensitivity, magical ideation and deviant olfactory experiences in 42 healthy subjects (21 Germans and 21 age- and gender-matched Japanese). The results show that: (1) Germans had significantly higher magical ideation scores and a higher frequency of deviant olfactory experiences than Japanese, and more Germans than Japanese reported having had deviant olfactory experiences at least once in their lives; (2) in Germans, the occurrence of deviant olfactory experiences was correlated with higher magical ideation scores; and (3) there was no relationship between olfactory sensitivity (olfactory thresholds) and either deviant olfactory experiences or magical ideation, respectively. We conclude that: (1) the lack of deviant olfactory experiences in Japanese may best be explained by cultural differences in the response attitude towards questionnaires requiring self-disclosure; (2) the positive relationship between magical ideation and deviant olfactory experiences strengthens the supposed link between 'psychotic-like' features in healthy populations and real hallucinations of psychiatric patients; and (3) the absence of a relationship between olfactory sensitivity and deviant olfactory experiences suggests that their anatomical-functional correlates within temporo-limbic regions may differ.