胰腺导管上皮细胞转分化为胰岛样细胞(英文)

背景:胰岛移植是治疗1型糖尿病包括部分2型糖尿病的有效方法。然而,供体来源的匮乏及移植免疫排斥反应极大地阻碍了该方法的推广和应用。最有希望的来源是从干细胞诱导分化出大量可供移植的胰岛细胞,但目前胰腺干细胞转分化方面的研究尚处于起步阶段。目的:寻找胰岛移植治疗糖尿病合适的细胞来源。方法:分离培养昆明小鼠胰腺导管上皮细胞,以添加有角化细胞生长因子、肝细胞生长因子和烟酰胺的DMEM/F12培养基培养,不同时间取样本于光镜和电镜下观察,检测第1,16天时CK-19、PDX-1免疫化学染色变化并以半定量RT-PCR检测第1,16天时胰岛素和胰高血糖素基因表达情况,21d时行双硫腙染色实验及葡萄糖刺激的胰岛素释放实验以检验胰岛样细胞的生理功能。结果与结论:分离第1天,大部分细胞CK-19染色阳性,偶可见PDX-1阳性细胞,16d后,CK-19阳性细胞快速增殖形成细胞团,大部分细胞PDX-1染色阳性;RT-PCR显示培养细胞胰岛素和胰高血糖素表达明显增强,分别增加了5.4倍和6.1倍(P0.01);21d时胰岛样细胞团更加成熟,双硫腙着色阳性,且对高糖(15mmol/L)刺激的胰岛素释放较低糖(5.6mmol/L)时增加了1.6倍(P0.05)。提示小鼠胰腺导管上皮细胞在体外培养条件下可增殖,并具有干细胞潜能,可转分化胰岛素分泌细胞。     

体外诱导胎儿骨髓间充质干细胞向胰岛β样分泌细胞的分化

背景：体外诱导人骨髓间充质干细胞定向分化为胰岛样细胞,目前尚无成熟的鉴定方案。 目的：探讨胎儿骨髓间充质干细胞在适当的条件下体外分化为胰岛样分泌细胞的可能性。   方法：将胎儿骨髓间充质干细胞与胎儿胰岛素细胞分别接种于Transwell 双层细胞培养板上下层共培养。对照组中的骨髓间充质干细胞仅用胰岛细胞培养基培养。倒置相差显微镜观察胎儿骨髓间充质干细胞、胎儿胰岛细胞的形态,放射免疫分析法检测共培养刺激下胰岛素的分泌情况。 结果与结论：未经诱导的骨髓间充质干细胞呈长梭形贴壁生长,与胰岛细胞共培养的骨髓间充质干细胞逐渐变圆,并聚集成团,共培养9 d后骨髓间充质干细胞经RAI检测分泌大量胰岛素,DTZ染色为阳性,细胞免疫化学检测阳性。而对照组无胰岛素释放。提示在适当的培养条件下,胎儿骨髓间充质干细胞具有向胰岛素分泌细胞分化的能力。     

兔脂肪干细胞体外诱导分化为胰岛β样细胞的可行性

背景：胚胎干细胞、胰腺干细胞、肝卵圆细胞、小肠上皮干细胞和骨髓间充质干细胞均可以在体外诱导分化为胰岛β样细胞,那么脂肪干细胞是否也可以呢？ 目的：探讨兔脂肪干细胞向胰岛β样细胞分化的可行性。 方法：以常规方法从兔脂肪抽吸物中分离、培养脂肪干细胞;传2代后,用高浓度葡萄糖(25 mmol/L)培养液DMEM以及碱性成纤维生长因子和尼克酰胺诱导兔脂肪干细胞向胰岛β样细胞分化。 结果与结论：第2代兔脂肪干细胞经过高糖诱导后,细胞形成细胞团;并且形成了双硫腙染色阳性的细胞团;免疫细胞化学表明诱导后细胞内的细胞胰岛素染色阳性。说明兔脂肪干细胞在体外可以定向诱导分化为胰岛β样细胞,这种胰岛β样细胞具有表达、储存胰岛素的功能。     

体外诱导胰腺干细胞向胰岛样细胞团的分化***☆

背景：目前由于胰岛来源匮乏,使得胰岛细胞移植治疗糖尿病无法满足临床需求,故体外将胰腺干细胞诱导分化为胰岛成为研究焦点。 目的：于体外将小鼠胰腺干细胞诱导成胰岛样细胞团并对其进行相关检测,探寻一种胰腺干细胞体外诱导分化成胰岛及鉴定的技术和方法。 方法：体外获得纯化的小鼠胰腺干细胞,采用联合诱导剂对其进行成胰岛方向的诱导分化,并对诱导形成的胰岛样细胞团进行形态学观察、双硫腙染色、RT-PCR和Western blot检测。 结果与结论：实验通过细胞形态学和细胞生长特性的观察以及免疫细胞化学染色证实体外成功获得了小鼠胰腺干细胞,采用联合诱导剂将其诱导成胰岛样结构,呈球形,以较细长的蒂部与瓶底连接,双硫腙染色将其染成铁红色。RT-PCR和Western blot 法可分别检测到胰岛样细胞团的胰岛素mRNA和胰岛素蛋白。结果证实小鼠胰腺干细胞可体外诱导分化成含β细胞的胰岛样细胞团。 关键词：胰腺干细胞;诱导;胰岛样细胞团;干细胞培养;小鼠 doi:10.3969/j.issn.1673-8225.2012.10.020     

人骨髓源干细胞向胰岛素分泌细胞分化(英文)

目的：探讨人骨髓源干细胞向具有功能的胰岛素分泌细胞分化的可能性。方法：从人骨髓分离间充质干细胞。采用表皮生长因子、β-巯基乙醇和高糖培养基诱导其向胰岛素分泌细胞分化。经诱导后,用RT-PCR检测胰岛β细胞相关基因的表达,并采用免疫细胞化学染色检测胞浆胰岛素的表达。此外,诱导后细胞分泌的胰岛素定量及胰岛素释放实验将采用化学发光法进行检测。将经诱导后的细胞移植到糖尿病小鼠的右侧肾被膜下。在移植后16 d持续检测小鼠的血糖水平,最后对右侧肾脏进行免疫组化检测。结果：经诱导后,细胞能表达胰岛β细胞相关基因;免疫细胞化学染色也能检测到胞浆有胰岛素的表达;而且这些细胞能对糖刺激有所反应。细胞被移植到糖尿病小鼠的肾被膜下,能起降血糖作用。其肾脏的免疫组化显示：肾被膜下有胰岛素阳性细胞。结论：人骨髓源干细胞具有向胰岛素分泌细胞分化的潜能,这将为糖尿病细胞治疗提供丰富的细胞来源。     

体外定向诱导人脐带间充质干细胞分化为胰岛样细胞

背景：人脐带间充质干细胞具有获取容易、免疫原性低等优点,目前尚缺乏体外诱导其分化为胰岛样细胞的成熟方案。 目的：探讨人脐带间充质干细胞在体外一定条件下分化为胰岛样细胞的可能性。 方法：无菌条件下分离培养人脐带间充质干细胞,细胞传3代后,采用两步法诱导其向胰岛样细胞分化：第1步,加入含100 μg/L β-神经生长因子,4 nmol/L ActivinA,10 mmol/L 尼克酰胺,25 μg/L表皮生长因子,体积分数为10%胎牛血清的DMEM/F12培养基中培养7 d;第2步,将诱导液换为含10 mmol/L尼克酰胺,10 μg/L碱性成纤维细胞生长因子,1%胰岛素-转铁蛋白-硒的DMEM/F12培养基,诱导时间为14 d。对照组单纯加入DMEM/F12培养基进行培养。 结果与结论：诱导2周后脐带间充质干细胞开始聚集成团,诱导3周后,葡萄糖刺激试验阳性、PDX-1和Insulin基因表达。而对照组细胞无胰岛素分泌,胰岛细胞相关基因表达阴性。实验成功地将脐带间充质干细胞诱导成了胰岛样细胞。     

β细胞素对胰腺干细胞转分化为胰岛样细胞团的影响

目的:探讨大鼠β细胞素在胰腺干细胞的增殖和转分化为胰岛细胞中的生物学作用.方法:分离并培养大鼠胰腺干细胞于CMRL1066和无血清DMEM/F12培养液中,不同浓度大鼠β细胞素处理后,双硫腙染色,放免法检测胰岛素分泌量.结果:在大鼠β细胞素浓度为20～30 mg/L时,大鼠胰腺干细胞转分化为胰岛样细胞团的数量明显高于对照组;其胰岛素分泌量明显高于对照组(P<0.01).结论:大鼠β细胞素具有促进胰腺干细胞转分化为胰岛β细胞并增强其胰岛素分泌的作用.     

人外周血单个核细胞向肝样细胞转化的体外诱导培养方法*

背景：外周血单个核细胞在体外条件下可向肝样细胞转化。 目的：探索体外诱导外周血单个核细胞向肝样细胞分化的培养方法。 方法：采用密度梯度离心法联合贴壁法纯化肝硬化患者外周血单个核细胞,以含巨噬细胞集落刺激因子、白细胞介素3及β-巯基乙醇培养基培养6 d后,诱导组用含肝细胞生长因子,成纤维细胞生长因子4的培养基培养14 d;以未诱导培养的单个核细胞及L02肝脏细胞系为对照。 结果与结论：外周血单个核细胞呈透明圆形、类圆形,用巨噬细胞集落刺激因子、白细胞介素3及 β-巯基乙醇培养基培养6 d后,部分细胞向成纤维样细胞生长,诱导后部分细胞呈多边形,多角形,14 d后细胞形态逐渐接近肝细胞,并表达白蛋白、甲胎蛋白、角蛋白18。说明在一定的诱导条件下,外周血单个核细胞在体外可以向肝样细胞分化。 关键词：肝样细胞;外周血;单个核细胞;诱导;分化;干细胞 doi:10.3969/j.issn.1673-8225.2012.10.018     

人脐静脉间充质干细胞向胰岛β样细胞诱导分化的实验研究

目的:探讨人脐静脉内皮及内皮下间充质干细胞向胰岛β样细胞分化的可行性。方法:采用胶原酶消化分离人脐静脉内皮及内皮下细胞进行贴壁培养;传代培养后,用高浓度葡萄糖(25mmol/l)培养液DMEM(含10?S)以及bFGF(basic Fibroblast Growth Factor)和尼克酰胺诱导脐静脉间充质干细胞向胰岛β样细胞分化。倒置显微镜下观察间充质干细胞诱导后形态变化,观察是否形成细胞团;用胰岛β细胞特异双硫腙染色鉴定诱导后细胞内是否含有高浓度游离锌离子;免疫细胞化学(Envision法)鉴定诱导后细胞是否分泌胰岛素。结果:第2代脐静脉间充质干细胞经过高糖诱导大约10天后,形成细胞团;呈双硫腙染色阳性;免疫细胞化学表明诱导后细胞内的细胞胰岛素染色阳性。结论:人脐静脉内皮及内皮下分离出的间充质干细胞在体外可以定向诱导分化为胰岛β样细胞,这种胰岛β样细胞具有胰岛素的分泌功能。     

不同细胞接种浓度影响人脐带间充质干细胞生长及多向分化

背景：在干细胞培养过程中寻找合理的细胞接种浓度,从而得到能稳定、高效生长的干细胞非常重要。 目的：观察不同接种浓度对人脐带间充质干细胞原代培养及分化为神经元样、胰岛样、脂肪样细胞的影响。 方法：分离人脐静脉内皮及内皮下层细胞,按不同接种浓度分组进行原代培养,记录原代培养时间并进行传代。取第1代人脐带间充质干细胞,体外诱导其向神经元样、胰岛样、脂肪样细胞分化。 结果与结论：原代培养发现以5×105~1×106/cm2细胞浓度接种的人脐带间充质干细胞生长状态最佳。经β-巯基乙醇诱导6 h,细胞即表达Nestin、神经丝蛋白及胶质纤维酸性蛋白;经尼克酰胺,活化素A,胰高血糖素样肽1诱导21 d可见明显的胰岛样细胞团,双硫腙染色呈砖红色;经成脂诱导培养基诱导后细胞油红O染色阳性,有明显脂滴出现。可见选取合适的细胞浓度可以快速获得大量的人脐带间充质干细胞,且该细胞经体外诱导可向神经元样、胰岛样及脂肪样细胞分化。关键词：细胞浓度;脐带间充质干细胞;原代培养;分化;神经元样细胞;胰岛样细胞;脂肪样细胞 缩略语注释：MSCs：mesenchymal stem cells,间充质干细胞 doi:10.3969/j.issn.1673-8225.2012.14.016     

大鼠胰腺导管干细胞分化形成类胰岛的改良诱导方法

目的 对大鼠胰腺导管干细胞（rPDSCs）分化形成类胰岛的诱导方法进行改良。 方法 在基础培养液DMEM/F12+10% FBS +1%青霉素/1%链霉素中,分别添加2、4、6和8 μmol/L全反式维A酸（ATRA）,体外诱导rPDSCs分化形成类胰岛,筛选ATRA最适诱导浓度。以ATRA最适诱导浓度为基础,再分别采用基质胶（matrigel）培养,悬浮培养或悬滴培养方式体外诱导rPDSCs分化形成类胰岛,筛选最适诱导培养方式。采用细胞形态学,双硫腙（DTZ）染色,细胞免疫荧光染色,Real-time PCR和ELISA方法对诱导类胰岛进行检测。 结果 与对照组相比,在基础培养液中,添加6 μmol/L ATRA及采用基质胶培养方式诱导效果最好。诱导28 d,细胞富集分化形成胰岛样球形细胞团;DTZ染色呈阳性;在基因和蛋白水平上分别表达胰岛素（insulin）和胰腺十二指肠同源框1（Pdx1）;葡萄糖刺激,释放胰岛素和C肽,且具有葡萄糖浓度依赖性。 结论 在基质胶培养方式下,采用6 μmol/L ATRA+DMEM/F12+10% FBS+1%青霉素/1%链霉素可以成功诱导rPDSCs体外分化形成类胰岛。     

骨髓间充质干细胞体外诱导分化为胰岛样细胞的降血糖作用

背景：移植胰岛及胰岛细胞治疗糖尿病已初见成效,但由于胰岛来源匮乏和免疫排斥反应而研究受阻。 目的：移植将大鼠骨髓间充质干细胞在体外诱导分化为胰岛样细胞,观察其对糖尿病大鼠的治疗作用。 方法：将大鼠骨髓间充质干细胞用碱性成纤维细胞生长因子、肝细胞生长因子等诱导,免疫细胞染色等检测诱导情况。SD大鼠腹腔注射链脲佐菌素建立糖尿病模型,建模成功后,随机分为对照组移植诱导胰岛样细胞的实验组,实验组经肾包囊移植诱导后的胰岛样细胞,对照组移植相同体积生理盐水,观察移植后糖尿病大鼠血糖和体质量变化。 结果与结论：大鼠骨髓间充质干细胞体外经肝细胞生长因子、碱性成纤维细胞生长因子等因子诱导后可以向胰岛样细胞转化。细胞移植后,对照组大鼠血糖无明显变化(P > 0.05),实验组大鼠血糖与对照组和移植前相比较,明显降低(P < 0.05)。大鼠骨髓间充质干细胞经含肝细胞生长因子、碱性成纤维细胞生长因子等的诱导体系可诱导成胰岛样细胞,经诱导的细胞有一定胰岛素分泌能力,将诱导后细胞通过肾包囊途径移植入糖尿病大鼠体内,可降低大鼠血糖水平。     

In vitro dedifferentiation of fetal porcine pancreatic tissue prior to transplantation as islet-like cell clusters

The fetal porcine pancreas under experimental conditions can be transplanted in the form of explants or islet-like cell clusters (ICCs) to normalize blood glucose levels in diabetic recipients. ICCs are released from the collagenase-digested pancreas and require a 4- to 5-day culture period for their complete formation. In order to maximize insulin producing beta cell differentiation following transplantation, an understanding of ICC development is essential to utilize this alternative treatment for type 1 diabetes. In this study a role is proposed for exocrine cells in the generation of the multipotent pancreatic precursor cells during the culture period. Acinar cells undergo dedifferentiation during the initial stages of the culture period into multipotent pancreatic precursor cells, previously called protodifferentiated cells. The progressive loss of exocrine differentiation appears to involve rapid degranulation of zymogen granules by exocytosis and loss of the prominent secretory apparatus. These processes occur in parallel with a significant reduction in the expression of lipase in the period from day 0 to day 5 and simultaneously there is an increase in the epithelioid/ductal cell marker, cytokeratin 20. Using proliferating cell nuclear antigen, cell proliferation during the culture period does not appear to account for the increase in epithelioid/ductal cells. Further the rates of apoptosis and necrosis which were identified using the TUNEL technique and propidium iodide, respectively, do not appear to account for the reduction in exocrine cell numbers. Exocrine cell dedifferentiation appears to increase the pool of protodifferentiated cells which have the potential to develop into the insulin-producing beta-cell population following transplantation into the diabetic recipient     

Conophylline与尼克酰胺联合诱导脂肪源干细胞为功能性类胰岛细胞

背景：1型糖尿病的胰岛移植治疗一直面临供体来源不足与免疫排斥两大关键问题,寻找一种自体来源的种子细胞通过组织工程方法制备类胰岛组织可以提供充足新型供体、降低异基因供体移植的不良反应。 目的：分析成人脂肪干细胞体外分化为对葡萄糖敏感、可分泌胰岛素的功能性胰岛样细胞团的能力,探索体外制备类胰岛组织的技术路线。 方法：首先分离纯化人体脂肪干细胞,采用新型植物诱导剂Conophylline与其他诱导因子的不同组合将脂肪干细胞诱导分化为胰岛样细胞团,观察不同组合的诱导分化效率,并利用特异性染色、RT-PCR,免疫细胞化学等方法对诱导分化后的细胞团在基因水平与蛋白水平上进行鉴定,最后用ELISA法检测细胞团在不同浓度葡萄糖刺激下胰岛素的分泌情况。 结果与结论：脂肪干细胞具有多能干细胞特性,可诱导分化为具有胰岛素分泌和葡萄糖浓度反应性类胰岛细胞团;Conophylline与尼克酰胺联合诱导可大幅度提高诱导分化效率。     

Chromatin-remodeling factors allow differentiation of bone marrow cells into insulin-producing cells

Type 1 diabetes is caused by the destruction of pancreatic beta-cells by T cells of the immune system. Islet transplantation is a promising therapy for diabetes mellitus. Bone marrow stem cells (BMSC) have the capacity to differentiate into various cell lineages including endocrine cells of the pancreas. To investigate the conditions that allow BMSC to differentiate into insulin-producing cells, a novel in vitro method was developed by using the histone deacetylase inhibitor, trichostatin A (TSA). BMSC, cultured in presence of TSA, differentiated into islet-like clusters under appropriate culture conditions. These islet-like clusters were similar to the cells of the islets of the pancreas. The islet-like clusters showed endocrine gene expression typical for pancreatic beta-cell development and function, such as insulin (I and II), glucagon, somatostatin, GLUT-2, pancreatic duodenal homeobox-1 (PDX-1), and Pax 4. Immunocytochemistry confirmed islet-like clusters contained pancreatic hormones. The colocalization of insulin and C-peptide was also observed. Enzyme-linked immunosorbent assay analysis demonstrated that insulin secretion was regulated by glucose. Western blot analysis demonstrated the presence of stored insulin. Electron microscopy of the islet-like cells revealed an ultrastructure similar to that of pancreatic beta-cells, which contain insulin granules within secretory vesicles. These findings suggest that histone-deacetylating agents could allow the differentiation of BMSC into insulin-producing beta-cells.     

Induced differentiation of human umbilical cord mesenchymal stem modified by cells Pdx1gene into islet beta-like cells in vitro

This study was to explore the induced differentiation of human mesenchymal stem cells (MSCs) modified by pancreatic and duodenal homeobox factor 1 (Pdx1) gene into insulin-producing cells in vitro. After recombined adenovirus vector with Pdx1 gene infected MSCs for 7 d, cells were induced by induction factors. The genes' expressions related to islet beta cells such as Pdx1, insulin, glucose transporter-2 (Glut2), were detected with RT-PCR, immunocytochemistry and Western blot. The levels of insulin and C peptide secretion were examined with chemiluminescence immunoassay. Insulin(+) cell rate was detected by flow cytometry. After infected by recombined adenovirus with Pdx1 and combined with induction factors, MSCs were aggregated and islet-like cell clusters formed. Dithizone staining of these cells was positive. The genes' expression related to islet beta cells, such as Pdx1, insulin, Glut2, could be detected. After induction, the islet-like cell clusters secreted insulin and C peptide. The levels of insulin and C peptide secretion increased with glucose stimulation. Insulin(+) cell rate was (11.61 +/- 4.83)%. It could be concluded that Pdx1 gene modified MSCs from human umbilical cord could be induced to differentiate into islet beta-like cells.     

The reversal of hyperglycaemia in diabetic mice using PLGA scaffolds seeded with islet-like cells derived from human embryonic stem cells

Islet-like cells derived from embryonic stem (ES) cells may be a promising therapeutic option for future diabetes treatment. Here, we demonstrated a five-stage protocol with adding exendin-4 instead of nicotinamide finally could generate islet-like cells from human embryonic stem (ES) cells. Immunofluorescence analysis revealed a high percentage of c-peptide positive cells in the derivation. However, in addition to insulin/c-peptide, most cells also coexpressed PDX-1 (pancreas duodenum homeobox-1), glucagon, somatostatin or pancreatic polypeptide. Insulin and other pancreatic beta-cell-specific genes were all present in the differentiated cells. Insulin secretion could be detected and increased significantly by adding KCL in high glucose concentration in vitro. Furthermore, subcutaneous transplantation of scaffolds seeded with the islet-like cells or cell transplantation under kidney capsules for further differentiation in vivo could improve 6 h fasted blood glucose levels and diabetic phenotypes in streptozotocin-induced diabetic SCID mice. More interestingly, blood vessels of host origin, characterized by mouse CD31 immunostaining, invaded the cell–scaffold complexes. This work reveals a five-stage protocol with adding exendin-4 may be an effective protocol on the differentiation of human ES cells into islet-like cells, and suggests scaffolds can serve as vehicles for islet-like cell transplantation.     

人脐带间充质干细胞分化胰岛样细胞过程中胰岛素和巢蛋白的表达

BACKGROUND:Domestic and international studies have confirmed that human umbilical cord mesenchymal stem cells could be induced to differentiate into islet-like cells, but little is reported about the changes of insulin and nestin expressions during the differentiation phase. OBJECTIVE:To observe the changes of insulin and nestin expressions during the differentiation of human umbilical cord mesenchymal stem cells into islet-like cells. METHODS:Human umbilical cord mesenchymal stem cells were cultured using UltraCULTURE medium in vitro. Stem cells were cultured for three generations to observe cell morphological changes under an inverted microscope, to test immunophenotype by flow cytometry, and to identify the capacity of osteogenesis and adipogenic differentiation. Induction protocol was divided into two stages. In stage 1, stem cells were induced for 14 days in the UltraCULTURE medium with 4 nmol/L activin A, 25 μg/L epidermal growth factor, 100 μg/L β-nerve growth factor, 10 mmol/L nicotinamide. In stage 2, the cells were cultured in the UltraCULTURE medium with 1% insulin-transferin-selenium, 10 mmol/L nicotinamide, 10 μg/L basic fibroblast growth factor for an additional 14 days. The expressions of nestin and insulin in those differentiated cells were tested by flow cytometry, and zinc ion expression in the islet-like cell clusters was identified by dithizone staining. RESULTS AND CONCLUSION:During the differentiation process, the insulin level was increased gradually in the induction group and reached a higher level on day 28, but the insulin expression showed negative in the control group. In addition, on day 14 of induced differentiation, the nestin expression reached the peak and then gradually reduced along with the prolonged inductive time. On day 28 of induction, islet-like cell clusters formed and were positive for dithizone staining. In this experiment, the umbilical cord mesenchymal stem cells were successfully induced and differentiated into islet-like cells, accompanied with the variation of insulin and nestin expression.     

胚胎干细胞来源的巢蛋白阳性细胞向胰岛素分泌细胞分化的实验研究

目的探讨胚胎干细胞向胰岛素分泌细胞分化的诱导，为糖尿病患者实施细胞移植治疗建立基础。方法将胚胎干细胞（ESC）在成纤维细胞（MEF）饲养层上扩增后脱离饲养层，让ESC自发分化成拟胚体（EB），再将EB诱导为巢蛋白（nestin）阳性的神经前体细胞（NPC），最后将NPC诱导成胰岛素分泌细胞（IPC）。结果ESC在MEF饲养层上扩增4d后，在悬浮培养状态下自发分化为EB。将EB在NPC选择性培养基中做贴壁培养后，ESC先分化为上皮样细胞，再分化为神经细胞样细胞。经过nestin免疫组化检测，在经过NPC培养基诱导4d的细胞，nestin阳性细胞占86．5％。nestin阳性细胞在IPC选择性培养基中诱导5～6d后，可分化成胰岛素分泌细胞。结论ESC来源的nestin阳性细胞既可以分化为NPC，也可以分化为IPC。