预诱导与全反式维甲酸并脑源性神经营养因子体外诱导骨髓基质干细胞向神经元样细胞的分化

背景：骨髓基质干细胞可定向诱导分化为神经元样细胞,但目前培养的方法尚不统一。 目的：采用预诱导与全反式维甲酸和脑源性神经营养因子为培养体系,拟在体外诱导大鼠骨髓基质干细胞向神经细胞分化。 设计、时间及地点：细胞学体外观察,于2007-12/2008-06在新疆医科大学第一附属医院医学研究中心干细胞室完成。 材料：清洁级雄性SD大鼠2只,由新疆医科大学动物试验中心提供。 方法：采用贴壁法体外分离培养大鼠骨髓基质干细胞,经反复传代细胞逐渐纯化。取生长状态良好的第3代骨髓基质干细胞,按8×107 L-1密度接种后,改换含10 μg/L碱性成纤维细胞生长因子、20 μg/L表皮生长因子的神经干细胞培养体系进行预诱导,48 h后去除预诱导液,加入含10 μg/L脑源性神经生长因子、1 μmol/L全反式维甲酸的神经干细胞培养体系定向诱导骨髓基质干细胞分化为神经细胞。以未诱导的骨髓基质干细胞作为对照。 主要观察指标：流式细胞仪检测第3代骨髓基质干细胞表面标志的表达,诱导后免疫荧光染色和流式细胞仪检测巢蛋白阳性的表达,免疫荧光染色鉴定神经元特异性烯醇化酶和胶质纤维酸性蛋白的表达。 结果：第3代骨髓基质干细胞CD29,CD44表达率分别为97.1%,99%,CD45表达率为0.5%,CD34呈阴性表达。预诱导48 h后巢蛋白阳性率为24%,而对照组仅为4.6%。诱导48 h后,巢蛋白染色呈阳性,并可见大量神经元特异性烯醇化酶和胶质纤维酸性蛋白阳性细胞出现;对照组均呈阴性表达。 结论：神经干细胞培养体系联合全反式维甲酸与脑源性神经生长因子,可在体外高效、稳定地诱导骨髓基质干细胞转化为神经元样细胞,并进一步向神经元和神经胶质细胞方向分化。     

无血清神经培养基体外培养大鼠骨髓源性神经干细胞

背景：传统的胚胎来源和脑来源的神经干细胞由于取材困难,并受伦理道德的约束,应用受到极大限制。 目的：拟利用无血清神经培养基体外培养大鼠骨髓源性神经干细胞。 方法：抽取大鼠股骨和胫骨的骨髓,采用全骨髓培养及贴壁筛选法分离培养骨髓间充质干细胞,流式细胞仪检测细胞周期及细胞免疫表型,油红O染色及茜素红染色鉴定其成骨、成脂能力。取传至4~6代的大鼠骨髓间充质干细胞,加入含表皮生长因子、碱性成纤维生长因子、B27的DMEM/F12无血清神经培养基进行诱导,采用免疫荧光染色及流式细胞仪予以鉴定。 结果与结论：P5骨髓间充质干细胞(91.5±3.1)%处于G1期,高表达CD90及CD29,不表达CD45及CD34,成脂诱导后在胞质中可见桔红色脂滴,成骨诱导后可见黑色矿化结节。骨髓源性神经干细胞巢蛋白免疫荧光染色呈阳性,流式细胞仪检测其阳性率为(97.2±1.1)%,NSE,β-Tubulin,GFAP及MAP-2抗原免疫荧光染色均呈阳性表达。表明在无血清神经培养基中加入特定生长因子,骨髓间充质干细胞可诱导为神经干细胞;在体外适当条件下,骨髓源性神经干细胞具有增殖和分化为神经元、星形胶质细胞及少突胶质细胞的能力。     

甲状腺激素对大鼠骨髓间充质干细胞分化为少突胶质细胞的影响

背景：如何为脱髓鞘疾病的细胞替代治疗提供丰富的少突胶质细胞来源是急需解决的问题。 目的：实验拟采用表皮生长因子及碱性成纤维细胞因子诱导骨髓间充质干细胞向神经干细胞方向分化,撤退细胞因子后,用甲状腺激素诱导神经干细胞向少突胶质细胞分化。 设计、时间及地点：细胞观察实验,于2007-08/12在泸州医学院神经生物学研究室完成。 材料：普通级SD大鼠5只用于骨髓间充质干细胞的培养。 方法：采用密度梯度离心法从大鼠骨髓中分离培养骨髓间充质干细胞,传至第4代,用含碱性成纤维细胞生长因子、表皮生长因子、N2辅助因子、甲状腺激素T3的DMEM/F12诱导液诱导向神经干细胞分化。诱导后4 d,更换成含胎牛血清、甲状腺激素T3的DMEM/F12分化液,诱导向少突胶质细胞分化。 主要观察指标：骨髓间充质干细胞生长情况和形态变化,采用SABC法进行免疫细胞化学检测神经细胞特异性标志的表达。 结果：原代细胞接种3 d后多数贴壁,传代后细胞贴壁速度加快,增殖能力更强。诱导液处理第4天,圆形细胞聚集成簇。换成分化液后,细胞伸出树枝状细长突起,交织成网,形成少突胶质细胞样细胞。骨髓源性细胞簇表达巢蛋白阳性,示神经干细胞;从细胞簇分化的细胞,多数细胞表达半乳糖脑苷脂,部分细胞表达髓鞘碱性蛋白,示少突胶质细胞,少数细胞表达微管相关蛋白2阳性,示神经元。 结论：甲状腺激素在体外可诱导骨髓间充质干细胞向少突胶质细胞分化。     

脂肪干细胞源性神经干细胞克隆球的电镜观察

目的观察脂肪干细胞(ADSCs)源性神经干细胞克隆球的超微结构形态。方法原代培养大鼠ADSCs,向神经干细胞诱导分化,分化为克隆球后,免疫荧光鉴定其Nestin表达,透射电子显微镜观察克隆球超微结构形态。结果大鼠ADSCs可诱导分化为细胞克隆球,其Nestin表达阳性,超微结构与原代培养的神经干细胞克隆球的超微结构相似。结论 ADSCs经诱导分化后形成的克隆球具有神经干细胞的特征。     

黄芩甙体外诱导大鼠骨髓基质细胞向神经干细胞的分化

背景：已有实验表明,骨髓基质细胞具有分化为骨骼、神经干细胞及造血干细胞的巨大潜能,而黄芩甙具有诱导细胞分化的作用。 目的：探讨黄芩甙体外诱导骨髓基质细胞分化为神经干细胞的可能性。 方法：分离培养Wistar纯系大鼠骨髓基质细胞,并将分离的骨髓基质细胞分为实验组和对照组,对照组不干预,实验组用黄芩甙(350~400 μmol/L)诱导,2组的培养环境相同。持续诱导6 d后选取生长良好的细胞进行蛋白质印迹和反转录PCR(RT-PCR)法检测。 结果与结论：黄芩甙诱导 7 d后,骨髓基质细胞形成较典型的神经细胞形态。诱导前骨髓基质细胞不表达神经细胞标记蛋白mRNA和蛋白;诱导 6 d表达神经细胞标记蛋白和mRNA;对照组不表达神经细胞标记蛋白和mRNA。证实黄芩甙在体外可诱导成年大鼠骨髓基质细胞分化为神经样细胞。     

大鼠骨髓基质细胞体外培养向神经干细胞的诱导分化

目的：已证实骨髓基质细胞可分化为中胚层组织细胞，实验予进一步探讨体外分离培养的骨髓基质细胞向神经干细胞分化的可能性，以及是否能继续定向分化为神经细胞及神经胶质细胞，为神经系统疾病细胞移植治疗提供种子细胞。 方法：实验于2007-02/09在泸州医学院神经生物学研究室进行。①动物：选择5只普通级SD大鼠，由泸州医学院实验动物中心提供，实验过程中对动物的处置符合动物伦理学标准。②实验方法：大鼠戊巴比妥钠腹腔麻醉，取双侧胫骨和股骨，磷酸盐缓冲液冲洗骨髓腔，采用密度梯度离心法从大鼠骨髓中分离培养骨髓基质细胞，胰蛋白酶与EDTA联合消化，传至第4代，用含20 μg/L碱性成纤维细胞生长因子、20 μg/L表皮生长因子、N2辅助因子的DMEM/F12培养液向神经干细胞诱导分化。③实验评估：观察原代、传代培养及诱导分化后的骨髓基质细胞生长情况和形态变化，采用SABC法进行免疫细胞化学检测神经细胞特异性标志的表达。 结果：①骨髓基质细胞形态观察：原代细胞接种1 d后开始贴壁增殖，3 d后多数贴壁，贴壁细胞呈梭形或扁平形；10 d后90%细胞融合，以长梭形为主，突起粗大，形成网状、片状；传代后细胞贴壁速度加快，增殖能力更强，7 d左右达到融合。②诱导分化后细胞生长情况和形态变化：第4代骨髓基质细胞向神经干细胞诱导分化7 d，成球的细胞脱离瓶底，悬浮在细胞液中。将细胞离心弃上清，换成血清分化液后，细胞球逐渐贴壁，球周围很快发出突起，分化为星形胶质细胞样细胞、神经元样细胞及少突胶质细胞样细胞。③神经细胞特异性标志的表达：骨髓源性细胞球表达巢蛋白，呈棕黄色，为神经干细胞；从细胞球分化的细胞抗胶质纤维酸性蛋白、微管相关蛋白2及半乳糖脑苷脂均呈阳性。 结论：骨髓基质细胞能在体外诱导分化出神经干细胞，且骨髓源性神经干细胞可进一步定向分化为神经细胞及神经胶质细胞。     

rhEGF和rhbFGF诱导胎鼠脑组织神经干细胞向神经元样细胞分化的研究

目的 观察在重组人表皮生长因子和重组人碱性纤维母细胞生长因子诱导下胎鼠脑组织神经干细胞向神经元样细胞分化的超微结构及细胞标志物表型特征.方法 采用孕16 d大鼠胚胎脑组织进行神经下细胞体外分离、培养传代及鉴定,分别于倒置相差显微镜及电子显微镜下观察经体外诱导培养后神经干细胞的组织形态及超微结构变化;免疫细胞化学染色检测神经微丝、微管相关蛋白-2、去甲肾上腺素、乙酰胆碱、γ-氨基丁酸以及多巴胺等蛋白质标志物的表达.结果 倒置相差显微镜观察显示,经体外培养的神经下细胞随着培养时间的延长逐渐形成神经干细胞球体,经体外诱导培养至第7天时即开始出现明显分化趋向,第14天时部分细胞分化发育完全;且蛋白质标志物神经微丝、微管相关蛋白-2、去甲肾上腺索、乙酰胆碱、γ-氨基丁酸及多巴胺均表达阳性.电子显微镜观察经体外诱导培养至第14天时,细胞出现明显的成熟分化趋向,细胞质中含有大量神经微丝和神经内分泌颗粒,呈典型的神经元分化特征.结论 神经千细胞在体外诱导培养条件下短期即可向神经元方向成熟分化.     

体外大鼠骨髓间充质干细胞多向分化潜能的研究

目的研究体外大鼠骨髓间充质干细胞向神经细胞等不同组织细胞多向分化的潜能。方法从SD大鼠股骨骨髓中获得间充质干细胞,原代培养后1∶2传代,传至第5代后分为普通传代培养组、神经细胞诱导组、成骨细胞诱导组和脂肪细胞诱导组。倒置相差显微镜下观察各组细胞生长情况、形态变化以及矿化结节和脂肪细胞的形成;流式细胞检测第5代大鼠骨髓间充质干细胞表面抗原CD29、CD44、CD90、CD31、CD34、CD45;免疫组织化学检测普通传代培养组和神经细胞诱导组细胞巢蛋白、微管相关蛋白-2、胶质纤维酸性蛋白和神经元特异性烯醇化酶等神经细胞相关蛋白的表达情况。结果大鼠骨髓间充质干细胞呈贴壁生长,细胞扩增至第5代时形态趋于一致,呈梭形。大鼠骨髓间充质干细胞表面抗原CD29(99.83%)、CD44(99.77%)、CD90(99.86%)均呈阳性表达,CD31(0.83%)、CD34(1.78%)、CD45(2.90%)无表达。在体外,普通传代培养细胞仅巢蛋白呈阳性表达;由神经细胞诱导的细胞巢蛋白、微管相关蛋白-2、胶质纤维酸性蛋白和神经元特异性烯醇化酶均呈阳性表达,且形态类似神经细胞;由成骨细胞诱导的细胞质内可见矿化结节形成;由脂肪细胞诱导的细胞质内出现多个猩红色呈簇状的脂肪滴。结论大鼠骨髓间充质干细胞易于提取、纯化和扩增,可于体外自发表达神经干细胞标志蛋白,并可通过诱导向神经细胞、成骨细胞及脂肪细胞分化。提示骨髓间充质干细胞不仅具有多向分化潜能,而且可能具有自发向神经干细胞分化的特性。     

骨髓间充质干细胞向甲状腺细胞的分化☆

背景：特定环境下体外可定向诱导大鼠骨髓间充质干细胞向甲状腺细胞分化。 目的：建立大鼠骨髓间充质干细胞体外诱导为甲状腺细胞的实验方法。 方法：采用密度梯度法分离培养大鼠骨髓间充质干细胞,应用促甲状腺素和胰岛素诱导,以未诱导组为平行对照。利用倒置光学显微镜、透射电镜观察细胞分化过程的形态学变化,免疫荧光等检测方法研究成人骨髓间充质干细胞的分化情况。 结果与结论：诱导培养第7天可见分化细胞中有甲状腺细胞的特有基因如TSHr的表达,第9天检测到分化细胞中甲状腺细胞标记物TTF-1的表达;对照组未见变化。形态学与生物学证实了骨髓间充质干细胞可诱导培养为甲状腺细胞。     

体外诱导人骨髓间充质干细胞向肝样细胞分化：肝细胞生长因子和表皮细胞生长因子的作用*

背景：研究证实人骨髓间充质干细胞能够诱导分化为肝样细胞,但其具体生物学特性及分化机制尚不清楚,且诱导分化培养体系尚不成熟。 目的：探讨肝细胞生长因子和表皮细胞生长因子体外诱导人骨髓间充质干细胞向肝样细胞分化的可行性。 方法：取食管癌手术患者肋骨,采用密度梯度离心联合贴壁筛选法获取人骨髓间充质干细胞,流式细胞仪检测细胞表面分子的表达。取第3代人骨髓间充质干细胞,分为4组,肝细胞生长因子组加入20 μg/L肝细胞生长因子,表皮细胞生长因子组加入20 μg/L表皮细胞生长因子,联合组同时加入上述两种生长因子,空白对照组不加任何生长因子。倒置显微镜下观察细胞形态的变化,诱导7,14 d RT-PCR检测甲胎蛋白与白蛋白mRNA的表达。 结果与结论：人骨髓间质干细胞不表达造血细胞标志CD34,CD45,强表达β1-整合素CD29和基质受体CD44。肝细胞生长因子组、表皮细胞生长因子组、联合组诱导后,细胞形态由长梭形变为类圆形或多角形,诱导第7,14天甲胎蛋白、白蛋白 mRNA均呈阳性表达;空白对照组未见多角形细胞,甲胎蛋白、白蛋白 mRNA均呈阴性表达。证明肝细胞生长因子、表皮细胞生长因子以及二者联合均具有诱导人骨髓间充质干细胞向肝样细胞分化的能力,至于二者联合是否能增强其分化能力尚待进一步免疫细胞化学染色定量分析予以验证。     

Immunocytochemical localization of three vesicular glutamate transporters in the cat retina

Vesicular transporters play an essential role in the packaging of glutamate for synaptic release and so are of particular importance in the retina, where glutamate serves as the neurotransmitter for photoreceptors, bipolar cells, and ganglion cells. In the present study, we have examined the distribution of the three known isoforms of vesicular glutamate transporter (VGLUT) in the cat retina. VGLUT1 was localized to all photoreceptor and bipolar cells, whereas VGLUT2 was found in ganglion cells. This basic pattern of complementary distribution for the two transporters among known populations of glutamatergic cells is similar to previous findings in the brain and spinal cord. However, the axon terminals of S-cone photoreceptors were found to express both VGLUT1 and VGLUT2 and some ganglion cells labeled for both VGLUT2 and VGLUT3. Such colocalizations suggest the existence of dual modes of regulation of vesicular glutamate transport in these neurons. Staining for VGLUT2 was also present in a small number of varicose processes, which were seen to ramify throughout the inner plexiform layer. These fibers may represent axon collaterals of ganglion cells. The most prominent site of VGLUT3 immunoreactivity was in a population of amacrine cells; the axon terminals of B-type horizontal cells were also labeled at their contacts with rod spherules. The presence of the VGLUT3 transporter at sites not otherwise implicated in glutamate release may indicate novel modes of glutamate signaling or additional roles for the transporter molecule.     

Distribution of monoamine-synthesizing neurons in the human medulla oblongata

We have employed immunohistochemical and morphometric procedures to study the distribution of monoamine-synthesizing neurons in the medulla oblongata of the adult human, utilizing antibodies to tyrosine hydroxylase (TH), phenylethanolamine N-methyltransferase (PNMT), and phenylalanine hydroxylase (PH8). In the human brain, the antigen with which PH8 reacts occurs within neurons that presumably synthesize serotonin (Haan et al., '87). Neurons containing these antigens were mapped and counted in successive coronal sections with the aid of a computer-assisted procedure. The results indicate that monoamine-synthesizing neurons are distributed in the human brain in patterns broadly similar to those described for other species. TH-immunoreactive cells extended caudorostrally for approximately 32 mm commencing at the spinomedullary junction and ending 8 mm caudal to the pontomedullary junction. In coronal sections these TH-immunoreactive neurons were seen in the lateral medulla dorsal to the inferior olive extending in a continuous band to the dorsomedial medulla. Above the obex the majority of these cells apparently synthesize adrenaline since many PNMT-immunoreactive cells were also found in this region. There were few or no PNMT-immunoreactive cells caudal to the obex, indicating that the TH-immunoreactive cells in this region synthesize either noradrenaline or dopamine. Approximately 65% of these TH-immunoreactive neurons contained melanin pigment, whereas few or no PNMT-immunoreactive cells contained melanin pigment. PH8-immunoreactive cells extended throughout the rostrocaudal extent of the medulla oblongata (approximately 40 mm). In coronal sections the majority were found in the medullary raphe nuclei. However, many cells throughout the rostrocaudal extent of the medulla were found laterally intermingled with catecholamine-synthesizing neurons. Occasional neurons in the lateral medulla appeared to contain both PH8- and TH-immunoreactivity.     

血管组织工程种子细胞在再生医学中的应用*☆◇

背景：小口径人工血管替代人体小动脉和静脉一直未获得满意的效果，因此研制出一种拥有较高远期通畅率的小口径人工血管成为了一个重要的研究课题。 目的：综述种子细胞在血管组织工程的研究进展。 方法：以 “Vascular tissue engineering, Seeding cells”为检索词，应用计算机检索Pubmed 数据库1960/2009有关文章。纳入有关血管组织工程种子细胞的文献。排除原始文献设计方法简单、结果可靠性差、非英文文献及结果重复的文献，保留35篇文献做进一步分析。 结果与结论：内皮细胞和平滑肌细胞是目前常用的种子细胞。内皮细胞和平滑肌细胞共同培养的体系，模拟体内环境，保持内皮细胞和平滑肌细胞具有正常的分泌功能和表型。骨髓间充质细胞可被有效的分离和扩增，在特定培养条件下可以诱导分化为多种血管细胞。在再生医学和生物组织工程方面有强大的潜力。     

Cellular and subcellular distribution of NMDA receptor subunit NR2B in the retina

Immunocytochemical studies showed the presence of staining for the N-methyl-D-aspartate (NMDA)-R2B glutamate receptor subunit at multiple sites in the cat retina. Reaction product in photoreceptor cells was localized at the inner/outer segment junction and in the axon terminals. Staining within the inner retina was limited to ganglion cells and their dendrites ramifying throughout the inner plexiform layer. These cells were seen to receive synaptic input from cone bipolar cells in both sublaminae. As with other glutamate receptor subunits, this immunoreactivity was typically confined to a single postsynaptic element at a cone bipolar dyad complex. Immunocytochemical localization of the NMDA-R1 subunit, considered to be an essential component of functional receptors, showed a widespread distribution across the retina including all the sites where NMDA-R2B staining was seen. Immunoprecipitation and Western blot analysis were used to confirm the presence of the NR2B receptor protein and its association with the NR1 subunit in both proximal and distal retinal layers. The findings suggest that NMDA-R2B subunits are positioned for multiple functions within the retina.     

周围神经组织工程中种子细胞的最新研究进展

典型的组织工程化人工神经主要包括种子细胞、支架材料以及有助于细胞生长、分化的细胞外基质，其中数量足够、不引起免疫排斥且有再生活力的种子细胞是其前提和基础。目前常用的种子细胞有雪旺氏细胞（SCs）、嗅球成鞘细胞（OECs）、骨髓基质细胞（BMSCs）和神经干细胞（NSCs）等。SCs一直是周围神经损伤修复研究领域的热点，OECs、BMSCs和NSCs修复周同神经损伤也在起步中。本从组织工程方面就上述四种细胞的最新研究进展进行综述，同时就存在的问题进行讨论。[第一段]     

Differential regulation of ciliary neurotrophic factor receptor-α expression in all major neuronal cell classes during development of the chick retina

Ciliary neurotrophic factor (CNTF) exerts a multiplicity of effects on a broad spectrum of target cells, including retinal neurons. To investigate how this functional complexity relates to the regulation of CNTF receptor α (CNTFRα) expression, we have studied the developmental expression of the receptor protein in chick retina by using immunocytochemistry. During the course of development, the receptor is expressed in all retinal layers, but three levels of specificity can be observed. First, the expression is regulated temporally with immunoreactivity observed in ganglion cells (embryonic day 8 [E8] to adult), photoreceptor precursors (E8–E12), amacrine cells (E10 to adult), bipolar cells (E12–E18), differentiated rods (E18 to adult), and horizontal cells (adult). Second, expression is restricted to distinct subpopulations of principal retinal neurons: preferentially, large ganglion cells; subpopulations of amacrine cells, including a particular type of cholinergic neuron; a distinctly located type of bipolar cell; and rod photoreceptors. Third, expression exhibits subcellular restriction: it is confined largely to dendrites in mature amacrine cells and is restricted entirely to outer segments in mature rods. These data correlate with CNTF effects on the survival of ganglion cells and mature photoreceptors, the in vitro differentiation of photoreceptor precursors and cholinergic amacrine cells, and the number of bipolar cells in culture described here or in previous studies. Thus, our results demonstrate an exceptional degree of complexity with respect to the regulation of neuronal CNTFRα expression in a defined model system. This suggests that the same signaling pathway is used to mediate a variety of regulatory influences, depending on the developmental stage and cell type. J. Comp. Neurol. 400:244–254, 1998. © 1998 Wiley-Liss, Inc.     

Distribution of neuropeptidelike immunoreactivities in the guinea pig olfactory bulb

The distribution of neuropeptidelike immunoreactivities in the adult guinea pig olfactory bulb was studied immunohistochemically with antisera raised against neurotensin (NT), substance P (SP), methionine-enkephalin-Arg6-Gly7-Leu8 (ENK), somatostatin (SOM), neuropeptide Y (NPY), and cholecystokinin-8 (CCK). In the main olfactory bulb, NT-like immunoreactive (NT-IR) neurons were found among periglomerular cells. In addition, a few periglomerular cells showed ENK-like immunoreactivity. Granule cells displaying SP- or ENK-like immunoreactivities and short axon cells with SOM- or NPY-like immunoreactivities were observed in the deeper half of the granule cell layer. SOM-IR short axon cells were also seen in the external plexiform layer. Dense NT- or NPY-IR fibers were distributed in superficial lamina of the granule cell layer, and sparse SP- or CCK-IR fibers were found in the glomerular layer. In the accessory olfactory bulb, some mitral, periglomerular, and granule cells showed NT-like immunoreactivity. SP- or ENK-IR granule cells were also observed. These results are discussed in relation to laminar organization of the olfactory bulb. The most characteristic features of peptide distribution in guinea pigs, as compared with that of rats in previous studies, were the relative abundance of NT-IR structures and the lack of SP- and CCK-IR juxtaglomerular and tufted cells.     

诱导性多潜能干细胞的应用前景

背景：通过基因转染的方式将已分化的体细胞重编程为诱导性多潜能干细胞,是近年来干细胞领域一项令人瞩目的新技术。由于诱导性多潜能干细胞摆脱了材料来源和伦理学的限制,因此其出现为特异的细胞治疗,特别是再生医学带来新的曙光。 目的：从诱导性多潜能干细胞的制备流程、产生的限制条件与机制、患者诱导性多潜能干细胞的获得及应用前景等方面做一评述。 方法：由第一作者检索PubMed数据库(www.ncbi.nlm.nih.gov/PubMed)2006-01/2010-03有关诱导性多潜能干细胞制备、产生机制的文章,检索词“induced pluripotent stem cells”,限定语言种类为English;同时手工检索部分文章。排除重复性研究,最终纳入34篇符合标准的文献。 结果与结论：诱导性多潜能干细胞系的建立主要包括以下几个步骤：①重组因子的选择。②目的细胞的选择。③重组因子的导入。④重组因子在目的细胞内的表达。⑤诱导性多潜能干细胞的产生。⑥重组细胞的鉴定。DNA甲基化、组蛋白的修饰作用和甲基化以及p53基因的表达在体细胞重编程为多潜能干细胞的过程中具有重要作用。虽然诱导性多潜能干细胞技术的研究仍然处于初级阶段,但是毫无疑问其具有广阔的应用前景。特别是患者特异性和疾病特异性诱导性多潜能干细胞的获得对于更好地理解疾病的发病机制及药物安全性评价提供了巨大的细胞种子资源。