人脐带源间充质干细胞静脉移植治疗大鼠肝纤维化

背景：近来国内外研究证明,来自其他组织的干细胞能够归巢到肝脏,并可能参与肝组织的再生,这为发展干细胞治疗肝脏疾病提供了新的希望。 目的：探究人脐带源间充质干细胞的分离、培养方法,观察脐带间充质干细胞移植对大鼠肝纤维化模型的修复作用,为脐带间充质干细胞的临床应用提供可靠的理论依据。 方法：自然贴壁法分离、纯化人脐带间充质干细胞并进行体外培养和扩增,用皮下多点注射CCl₄制备肝纤维化大鼠模型。将22只模型大鼠随机分为模型损伤组(n=11)和细胞移植组(n=11)。细胞移植组在模型制备成功后的第1,2,3周经尾静脉给予1×10⁶脐带间充质干细胞治疗,4周后将大鼠处死,收集各组大鼠血液检测肝功能;摘取肝脏行苏木精-伊红染色,观察病理变化;免疫组化法观察库普弗细胞的数量及分布;免疫组化法观察治疗组脐带间充质干细胞的定位情况。 结果与结论：脐带间充质干细胞经尾静脉移植入肝硬化大鼠后,大鼠的肝功能均明显改善,与对照组比较,差异有显著性意义(P < 0.05);肝组织苏木精-伊红染色提示,肝纤维化程度明显改善;免疫组化法观察库普弗细胞的数量提示,库普弗细胞数量明显减少;免疫组化方法利用抗BrdU抗体在治疗组大鼠肝脏观察到BrdU标记的脐带间充质干细胞。说明脐带间充质干细胞移植可以改善大鼠外周血液的血生化特性和肝的组织学结构。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

人脐带源间充质干细胞移植改善四氯化碳诱导肝硬化大鼠的肝纤维化*☆

背景：干细胞移植作为一种全新的肝硬化治疗方法的可行性和有效性,在国内外文献中报道极少。 目的：观察人脐带源间充质干细胞移植对四氯化碳诱导肝硬化大鼠肝纤维化的影响。 方法：32只Wistar大鼠随机分为2组,对照组经尾静脉内注射生理盐水,肝硬化组采用四氯化碳植物油皮下注射8周制成肝硬化大鼠模型,再随机分为3组,盐水对照组、人脐带源间充质干细胞移植组分别经尾静脉内注射生理盐水和人脐带源间充质干细胞混悬液,8周肝硬化组无其他干预。 结果与结论：对照组血清碱性磷酸酶水平明显低于其他3组(P < 0.05);人脐带源间充质干细胞移植组血清白蛋白和胆固醇水平与对照鼠相近(P > 0.05),与8周肝硬化组和盐水对照组相比明显升高(P < 0.05)。血清碱性磷酸酶水平也出现了明显下降(P < 0.05)。肝硬化8周组大鼠肝组织中有大量胶原纤维增生并形成假小叶;盐水对照组与肝硬化8周组相似;仅在人脐带源间充质干细胞移植组大鼠肝中观察到散在的绿色抗人抗核抗体阳性细胞,肝组织中胶原纤维的量明显小于盐水对照组。提示经尾静脉移植人脐带源间充质干细胞,可明显改善四氯化碳诱导肝硬化大鼠的肝纤维化程度。 关键词：脐带源间充质干细胞;细胞移植;肝硬化;四氯化碳;大鼠 doi:10.3969/j.issn.1673-8225.2012.10.028     

重复测量分析脐带间充质干细胞移植治疗肝硬化的疗效

背景：国内外已有应用间充质干细胞治疗肝硬化的临床研究且取得了可喜的进步,但在实际应用中,经常会误用t 检验分析这类资料。 目的：采用重复测量方法分析脐带间充质干细胞治疗肝硬化患者的有效性和安全性。 方法：失代偿性肝硬化患者27例,均行常规内科治疗,包括护肝、对症治疗等。在入院1周后静脉移植脐带间充质干细胞(第2-4代),细胞存活率≥90%,干细胞数量≥2×10⁷个,共治疗4次,每次间隔5-7 d。使用重复测量方差分析脐带间充质干细胞移植治疗不同时点的肝功能变化。 结果与结论：单变量重复测量方差分析结果显示,在治疗后2,3个月时血清白蛋白升高、总胆红素降低,与治疗前比较差异有显著性意义(P < 0.05);治疗后3个月,谷草转氨酶降低、胆碱酯酶升高,与治疗前比较差异有显著性意义(P < 0.05),所有患者在观察期内无肝脏及其他器官肿瘤发生。结果表明脐带间充质干细胞移植治疗肝硬化安全有效,患者肝功能得到改善。     

脐带间充质干细胞移植治疗终末期肝硬化的疗效及安全性

背景：骨髓间充质干细胞移植已应用于治疗终末期肝病并取得一定疗效。 目的：观察脐带间充质干细胞治疗终末期肝硬化患者的疗效及安全性。 方法：无菌条件下分离培养脐带间充质干细胞,选择60例肝硬化患者,在内科治疗基础上经肝动脉插管进行脐带间充质干细胞移植。 结果与结论：移植2,4,8,12周后,患者血清白蛋白、前白蛋白水平逐渐升高(P < 0.05),总胆红素、凝血酶原时间明显低于治疗前水平(P < 0.05)。移植后患者乏力、腹胀、纳差明显好转,未发生与移植相关的严重并发症。说明脐带间充质干细胞治疗终末期肝硬化安全有效,能改善患者的肝功能、凝血功能及临床症状。     

骨髓间充质干细胞经尾静脉移植治疗肝纤维化

BACKGROUND: Liver fibrosis is the early stage of terminal liver diseases. Effective treatment for liver fibrosis can prevent the occurrence of terminal liver diseases. Bone marrow mesenchymal stem cell transplantation is a promising method to treat liver fibrosis. OBJECTIVE: To study the therapeutic effect of bone marrow mesenchymal stem cells on liver fibrosis in rats. METHODS: Eighteen Sprague-Dawely rats were randomized into three groups: control, model and cell transplantation groups. Animal models of carbon tetrachloride-induced liver fibrosis were made in the latter two groups. After modeling, 1 mL bone marrow mesenchymal stem cells (5×105) or the same volume of normal saline was injected via the tail vein into the rats in the cell transplantation and model groups, respectively. Rats in the control group were given no treatment. Degree of liver fibrosis, liver function, histological changes of the liver were detected and observed in the three groups at 4 weeks after treatment. RESULTS AND CONCLUSION: In the control group, the liver tissues had normal structure with no fibrosis; in the model group, proliferation of fibrous tissues in the portal area of the liver, inflammatory cell infiltration, vacuolar degeneration and irregular arrangement of liver cells, and tissue structure damage were observed; in the transplantation group, liver tissue damage was severer than the control group but milder than the model group. Levels of serum hyaluronidase, type IV collagen and procollagen III were significantly lower in the cell transplantation group than the model group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cell transplantation can alleviate liver fibrosis and improve liver function in rats with carbon tetrachloride-induced liver fibrosis.  中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程      

骨髓间充质干细胞移植治疗肝硬化的研究进展

肝硬化是临床中常见的慢性进行性肝病,目前治疗晚期肝硬化最有效的方法是肝脏移植,但肝源缺乏、费用昂贵、移植排斥反应及长期应用免疫抑制剂引起并发症等成为限制其广泛应用的主要原因.干细胞移植有利于受损肝组织修复,能够代偿部分肝功能,已成为治疗肝病的一种新方法.就骨髓间充质干细胞移植治疗肝硬化的基础、临床研究进展、存在的问题以及发展前景作一综述,旨在为其进一步研究提供理论依据.     

人脐带间充质干细胞移植肝硬化大鼠肝脏miRNA的差异表达

背景：研究表明脐带间充质干细胞可以显著改善肝硬化的程度,进而修复肝损伤,但其治疗肝硬化的分子调控机制,尤其是非编码RNA调控的肝内基因变化,目前并没有得到详细的阐释。 目的：分析人脐带间充质干细胞移植肝硬化大鼠肝细胞中微小RNA基因表达的变化。 方法：采用四氯化碳皮下注射联合乙醇喂服方法建立肝硬化大鼠模型,造模8周后经尾静脉输注人脐带间充质干细胞,每周1次,连续注射4周。最后一次注射治疗1周后收集大鼠肝脏组织进行石蜡切片和提取肝脏RNA进行表达谱基因芯片分析,同时收集血清利用自动生化分析仪测定肝功能指标变化。 结果与结论：人脐带间充质干细胞治疗可以显著降低谷丙转氨酶、谷草转氨酶和转肽酶水平,脂肪病变和肝细胞坏死显著减少。微小RNA表达谱芯片杂交分析和PCR验证结果显示rno-miR-369-5p、rno-miR-3584-5p和rno-miR-153*这3种微小RNA基因在造模过程中先下调表达,并在人脐带间充质干细胞治疗后显著上调;而rno-miR-93、rno-miR-199a-3p、rno-miR-195、rno-let-7a和rno-miR-19a这5种微小RNA基因在造模过程中先上调表达,并在人脐带间充质干细胞治疗后显著下调。以上结果表明人脐带间充质干细胞逆转肝硬化和肝细胞损伤过程中,可能通过上调rno-miR-369-5p、rno-miR-3584-5p和rno-miR-153*等miRNA基因表达,下调rno-miR-93、rno-miR-199a-3p、rno-miR-195、rno-let-7a和rno-miR-19a等相关miRNA基因表达发挥治疗作用。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

人脐带间充质干细胞治疗失代偿期肝硬化的近期效果*

背景：脐带间充质干细胞可分化为肝细胞样细胞。 目的：观察人脐带间充质干细胞移植治疗失代偿期肝硬化的近期临床效果。 方法：选择肝硬化失代偿期患者30例,对照组18例给予内科常规治疗,移植组12例在内科综合治疗基础上经肝动脉移植人脐带间充质干细胞悬液治疗。 结果与结论：移植后,移植组白蛋白有上升趋势,从第2周开始明显高于对照组(P < 0.05),并持续至第12周;谷丙转氨酶和凝血酶原时间有下降趋势,从第8周开始明显低于对照组(P < 0.05),并持续至第12周;总胆红素水平有下降趋势,从第4周开始明显低于对照组(P < 0.05),并持续至第12周。两组临床症状、体征改善情况差异无显著性意义(P > 0.05)。说明经股动脉移植人脐带间充质干细胞是治疗失代偿期肝硬化近期有效。关键词：肝硬化;脐带;间充质干细胞;失代偿期;肝动脉;移植 缩略语注释：UC-MSCs：human umbilical cord mesenchymal stem cells,人脐带间充质干细胞 doi:10.3969/j.issn.1673-8225.2012.14.024     

脐带间充质干细胞移植治疗系统性红斑狼疮

<正>据南京大学医学院附属鼓楼医院2014年5月25日[Arthritis Res Ther,2014,16(2):R79.]报道,南京大学医学院附属鼓楼医院、江苏大学附属医院及苏北人民医院、江苏省人民医院等多家研究中心联合进行了一项用脐带间充干细胞移植治疗严重系统性红斑狼疮(SLE)的多中心临床试验。结果表明,脐带间充质干细胞移植能够对SLE患者产生满意的临床疗效。SLE是一种常见的和潜在致命的自身免疫性疾病。其特征在于机体产生大量抗体,并与体内相应的自身抗原结合形成相应的免疫复合物,对肾脏、心血管、神经、肌肉骨骼和皮肤产生系统损害。     

脐带间充质干细胞移植治疗大鼠糖尿病

背景：脐血间充质干细胞具有很强的增殖能力和分化能力,在趋向分化作用下可以分化成胰岛β细胞,进而起到治疗糖尿病的作用。 目的：观察移植脐带间充质干细胞对大鼠糖尿病的治疗效果。 方法：30只雄性SD大鼠中随机取6只作为对照组,注射生理盐水;其中24只按45 mg/kg的剂量注射链脲霉素建立糖尿病模型后,随机等分为移植组和糖尿病组,移植组大鼠尾静脉注射移植脐带间充质干细胞。 结果与结论：造模后30 d,糖尿病组大鼠空腹血糖维持在较高水平,且高于对照组(P < 0.05)。造模后,与糖尿病组相比,移植组大鼠空腹血糖水平显著下降(P < 0.05),体质量显著增加(P < 0.01),45 d时移植组大鼠空腹血糖水平与体质量接近对照组水平(P > 0.05),而糖尿病组大鼠空腹血糖维持较高水平,且体质量持续下降。提示脐带间充质干细胞移植能有效治疗大鼠糖尿病。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

人端粒酶反转录酶基因修饰脐带间充质干细胞移植治疗急性肾损伤

背景：人端粒酶反转录酶(hTERT)是调控增殖及定向分化的首选生长因子之一,具有多重生物学效应,为建立基因工程的永生化干细胞系奠定了基础。 目的：探讨人端粒酶反转录酶基因修饰脐带间充质干细胞移植对大鼠缺血再灌注诱导的急性肾损伤的治疗作用。 方法：体外培养人脐带间充质干细胞,构建缺血再灌注诱导的大鼠急性肾损伤模型,建模后将大鼠随机分为3组：对照组尾静脉注射1 mL L-DMEM培养液;空载病毒组：尾静脉注射1 mL经空载病毒转染人脐带间充质干细胞悬液;hTERT转染组尾静脉注射1 mL经PLXSN-hTERT转染的人脐带间充质干细胞悬液。 结果与结论：移植后第3,28天苏木精-伊红染色检查示hTERT转染组的肾小管损伤评分<空载病毒组<对照组(P < 0.05)。移植后第28天,CM-Dil 阳性细胞数为hTERT转染组>空载病毒组>对照组(P < 0.05)。移植细胞后第1,3,14,28天血肌酐、尿素氮水平均为hTERT转染组<空载病毒组<对照组(P < 0.05)。结果证实,hTERT基因修饰脐带间充质干细胞移植对大鼠急性肾损伤具有明显的修复作用。  中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

人羊膜间充质干细胞移植修复肝脏缺血-再灌注损伤

BACKGROUND:Studies have shown that human amniotic mesenchymal stem cells can differentiate into hepatocyte-like cells, suggesting that human amniotic mesenchymal stem cell transplantation provides a new potential for the clinical treatment of liver diseases. OBJECTIVE:To observe the effect of human amniotic mesenchymal stem cell transplantation on the repair of liver ischemia-reperfusion injury repair. METHODS:Sixty Sprague-Dawley rats were randomized into stem cell transplantation, model and control groups. Animal models of liver ischemia-reperfusion injury were made in the rats in the stem cell transplantation and model groups. One hour after modeling, rats in the stem cell transplantation were given injection of human amniotic mesenchymal stem cells (0.5 mL, 106 cells) via the tail vein, while rats in the model and control group were given L-DMEM (0.5 mL) or normal saline (0.5 mL), respectively. Liver function and liver morphology were detected at 1, 2, 3 weeks after transplantation. Meanwhile, RT-PCR detection and western blot assay were also conducted. RESULTS AND CONCLUSION:(1) Liver function: Compared with the control group, levels of aspartate aminotransferase, alanine aminotransferase and malondialdehyde were significantly increased in the model group at different time points after transplantation (P < 0.05), while a significant reduction in the levels of these three indicators was found after cell transplantation as compared with the model group (P < 0.05). (2) Liver morphology: 2 weeks after transplantation, rats in the model group exhibited hepatocyte degeneration and necrosis, and severe fibrosis, but these changes were remarkably alleviated in the stem cell transplantation group. (3) PT-PCR and western blot detection: 2 weeks after transplantation, a significantly higher level of hepatocyte growth factor in the liver tissue and a lower level of α-smooth muscle protein were found in the stem cell transplantation group compared with the model group (P < 0.05). All these experimental findings indicate that human amniotic mesenchymal stem cell transplantation can improve impaired liver function in rats, possibly through regulating hepatocyte growth factor and α-smooth muscle protein expression levels in the liver, and thereby promotes the repair of liver ischemia-reperfusion injury.     

脐带间充质干细胞移植对不同证型系统性红斑狼疮患者的效果分析

背景：系统性红斑狼疮按中医辨证分为热毒炽盛等4个症型,治疗以补肾养阴,清化淤毒为主,但仍有许多患者治疗效果不佳。间充质干细胞具有多向分化、造血支持和免疫调节的功能,目前已有多项研究用于治疗难治性、复发性系统性红斑狼疮,取得良好疗效。目的：探讨脐带间充质干细胞移植对不同证型系统性红斑狼疮患者的疗效。方法：系统性红斑狼疮患者21例,经中医辨证分成热毒炽盛证、肝肾阴虚证、脾肾阳虚证及气滞血瘀证4型,分别统计患者脐带间充质干细胞移植前后各型的临床及实验室指标变化。结果与结论：脐带间充质干细胞移植1,3,6个月均可有效减少系统性红斑狼疮患者实验尿蛋白含量,降低系统性红斑狼疮疾病活动指数评分(P < 0.01)。与移植前比较,移植脐带间充质干细胞后1,3,6个月可显著减少肝肾阴虚型患者尿蛋白含量(P < 0.01),移植后1,3个月对热毒炽盛及气滞血瘀型尿蛋白含量为轻度减少(P < 0.05),脾肾阳虚型患者在植后1个月时的尿蛋白含量轻度减少(P < 0.05)。脐带间充质干细胞移植可提高所有中医分型患者的血浆白蛋白含量(P < 0.01),其中对热毒炽盛型效果稍差(P < 0.05)。脐带间充质干细胞移植后各型患者外周血血小板有上升趋势,但与移植前相比差异无显著性意义。结果说明,脐带间充质干细胞移植治疗系统性红斑狼疮有效,对不同症型的系统性红斑狼疮患者的疗效有一定的差异。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

脂肪间充质干细胞移植后肝硬化大鼠血生化指标的变化

BACKGROUND:Stem cell transplantation is a promising treatment of advanced liver disease, and adipose-derived mesenchymal stem cells have become another kind of popular cells following bone marrow mesenchymal stem cells. OBJECTIVE:To investigate the influence of adipose-derived mesenchymal stem cell transplantation on blood biochemical indices of liver cirrhosis rats. METHODS:Sixty rats were equally randomized into normal control, model and cell transplantation groups. Model rats of liver cirrhosis were made in the latter two groups through intragastric administration of carbon tetrachloride. One week after successful modeling, rats were given intraperitoneal injection of adipose-derived mesenchymal stem cell suspension in the cell transplantation group, and given normal saline in the other two groups. RESULTS AND CONCLUSION:Compared with the normal control group, the model group showed a significant increase in the levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, total protein in liver tissues, serum level of malondialdehyde, 15-minute indocyanine green retention rate and degree of hepatic fibrosis, and a significant decrease in serum albumin level, serum albumin/globulin, levels of glutathione peroxidase and cyclic guanosine monophosphate in liver tissues. On the contrary, these indicators were all improved in the cell transplantation group compared with the model group. Moreover, CM-Dil-positive cells were visible in the liver tissue of rats undergoing adipose-derived mesenchymal stem cell transplantation. All these findings indicate that adipose-derived mesenchymal stem cell transplantation can reduce liver cirrhosis in rats by acting on blood biochemistry levels.     

不同预处理策略促进间充质干细胞的归巢

BACKGROUND:Homing is the initial and key procedure of stem cells-based tissue restoration. Current studies have shown that the inability to recruit bone marrow mesenchymal stem cells to target tissue with high efficiency remains a significant barrier to tissue restoration. Preconditioning strategy provides a new insight to promote stem cell homing. OBJECTIVE:To review preconditioning strategies for promoting the homing of stem cells. METHODS:In PubMed database, different combinations of terms from “stem cell, mesenchymal stem cells, preconditioning, homing, migration” served as search terms to retrieve articles referring preconditioning strategies for promoting mesenchymal stem cell homing published from January 2000 to September 2015. According to the inclusion criteria, 72 articles were selected for final review. RESULTS AND CONCLUSION:Pretreating target tissue or mesenchymal stem cells ahead of cell transplantation, known as tissue preconditioning or cell preconditioning, prominently promotes the homing of mesenchymal stem cells, therefore enhancing tissue restoration effect. Tissue preconditioning is designed to up-regulate expression of chemokines by varying the local microenvironment, thereby increasing homing ability of mesechymal stem cells. Mesenchymal stem cell preconditioning strategies, for example, gene modification and cytokine induction, are mainly to up-regulate expression of chemokine receptors on the surface of mesenchymal stem cells as effectors, and thus promote targeted cell homing. Overall, preconditioning strategy will bring great hope to apply stem cell therapy into the clinic.     

在5-杂氮胞苷诱导下人脐带间充质干细胞生物活性特性及向心肌样细胞的分化

BACKGROUND: Human umbilical cord mesenchymal stem cells belong to a special class of stem cells with a limited number, which are difficult to isolate and purify. Importantly, there is a lack of clinical understanding of biological characteristics of human umbilical cord mesenchymal stem cells.     

经肝动脉途径兔自体骨髓干细胞移植治疗肝硬化

BACKGROUND:How to make more transplanted bone marrow stem cells stay and differentiate in the liver is an important issue, which is also crucial for treatment of liver cirrhosis via the hepatic artery. OBJECTIVE:To investigate the therapeutic effect of autologous bone marrow stem cell transplantation via the hepatic artery on liver cirrhosis. METHODS:Thirty New Zealand white rabbits were equivalently randomized into normal control, stem cell transplantation and model groups. Animal models of liver cirrhosis were made in the latter two groups. Then, model rabbits in the stem cell transplantation group were subjected to autologous bone marrow stem cell transplantation via the hepatic artery. Liver function of rabbits was detected in 1, 2, 4, 8, 10 weeks after cell transplantation, and pathological detection of the liver was performed in the 10th week. RESULTS AND CONCLUSION:At 10 weeks after cell transplantation, the liver function of the rabbits was improved significantly compared with the model group, including reduced activities of serum alanine aminotransferase, total bilirubin and aspartate aminotransferase, shortened activated partial thromboplastin time, and increased albumin level (P < 0.05). Pathological examination of the liver showed that the liver cells in the stem cell transplantation group were intact with no obvious edema and still had the structure of the pseudolobule, and compared with the model group, the degree of liver fibrosis was significantly reduced in the stem cell transplantation group. Our experimental results show that the transplantation of autologous bone marrow stem cells via the hepatic artery has a certain therapeutic effect on liver cirrhosis by increasing the body albumin content in a short time and improving the liver function.     

三维培养诱导人脂肪间充质干细胞微球的成软骨分化能力

背景：关节软骨损伤后修复结果不满意,需要新的手段,而脂肪间充质干细胞较适宜做种子细胞诱导软骨,然而怎么能够使诱导的软骨具有功能需要研究。 目的：采用三维培养体系诱导人脂肪间充质干细胞微球向软骨分化。 方法：无菌切取吸脂术后脂肪组织,分离培养人脂肪间充质干细胞,传至第3代进行流式细胞术分析,成骨成脂肪诱导等鉴定,同时也给予合适的培养条件用三维培养的方式向软骨细胞诱导,并行阿利辛蓝染色鉴定糖胺多糖的合成,苏木精-伊红染色进行组织学分析,免疫荧光检测Ⅱ型胶原表达,称质量测量软骨硬度。 结果与结论：分离的人脂肪间充质干细胞CD105,CD44,CD29均高表达,而 CD45,CD34低表达,并且成骨成脂诱导后细胞茜素红染色和油红O染色均为阳性。三维培养法诱导的软骨细胞可表达大量糖胺多糖及Ⅱ型胶原。结果证实,三维培养法诱导人脂肪间充质细胞向软骨分化后,具有软骨细胞的特性。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

调控脂肪间充质干细胞成软骨分化基因Sox-9和低氧诱导因子1α的表达

BACKGROUND:Gene transfection of adipose-derived mesenchymal stem cells (ADSCs) can stably synthesize seed cells that secrete specific cytokines, thereby achieving long-term stable proliferation, differentiation and chondrogenic differentiation of seed cells. OBJECTIVE:To observe the effect of insulin-like growth factor-1 (IGF-1) gene transfection on the expression of Sox-9 and hypoxia-inducible factor-1alpha (HIF-1α) in rabbit ADSCs. METHODS:ADSCs were harvested from the posterior subcutaneous adipose tissue from Adult New Zealand white rabbits, then the cells were identified by immune fluorescent assay. After being transfected with pcDNA3.1-IGF-1, the cells were selected with G418. RT- PCR and western blot were used to analyze the expression level of HIF-1α in normal ADSCs, pcDNA3.1-transfected ADSCs, and pcDNA3.1-IGF-1- transfected ADSCs. The growth curve of cells was measured by MTT, and meanwhile, proliferation efficiency of cells was measured by counting CM-DiL-labeled cells. Immune fluorescent assay was used to analyze the expression of HIF-1α and Sox-9. And the content of glycosaminoglycan in each group was determined by using dimethylmethylene blue dye-binding assay. RESULTS AND CONCLUSION:The stably expressed pcDNA3.1-IGF-1 cell lines were successfully established and showed stable expression of IGF-1 at mRNA and protein levels. MTT and CM-Dil fluorescence results suggested that IGF-1-transfected cells displayed stronger proliferation ability and efficiency, and these cells showed a positive expression of HIF-1α and Sox-9 as well as higher glycosaminoglycan content than the other two groups (P < 0.01). All these findings indicate that IGF-1 gene transfection can effectively promote the proliferation of ADSCs, and promote the positive expression of Sox-9 and HIF-1α. And the secretion of chondral extracellular matrix such as glycosaminoglycan is also significantly improved.     

人脐带间充质干细胞移植治疗急性肺损伤

BACKGROUND:Studies have shown that human umbilical cord mesenchymal stem cells can improve pulmonary ventilation function by reducing inflammations. OBJECTIVE:To observe the therapeutic effect of human umbilical cord mesenchymal stem cell transplantation on acute lung injury. METHODS:Thirty Sprague-Dawley rats were randomized into normal group, model group and experimental group. Rats in the latter two groups were used to establish animal models of acute lung injury by intratracheal instillation of lipopolysaccharide. One hour after modeling, rats in the experimental group were intratracheally administered human umbilical cord mesenchymal stem cell suspension (0.1 mL, 1×10⁶ cells), and those in the other two groups were given normal saline in the same dose intratracheally. Twenty-four hours after treatment, the pathological changes of lung tissue were observed using hematoxylin-eosin staining; the wet and dry weight ratio of the lung tissue and the levels of serum interleukin-1 and interleukin-8 were detected. RESULTS AND CONCLUSION:Compared with the normal group, the wet and dry weight ratio of the lung tissue and the levels of serum interleukin-1 and interleukin-8 were significantly increased in the model group (P < 0.05), while compared with the model group, these levels were significantly decreased in the experimental group (P < 0.05). Hematoxylin-eosin staining results showed clear alveolar space structure with complete alveolar septum in the normal group. In the model group, the alveolar septum was markedly thickened, and there was visible pulmonary capillary hyperemia, edema, as well as a large amount of inflammatory cell infiltrations in the pulmonary capillaries and alveolar space. Edema fluid rich in proteins was observed in a part of the pulmonary alveoli, and an extensive transparent membrane formed in the alveolar space. In the experimental group, the alveolar structure was clear, but the alveolar septum became thickened, and red blood cells and a small amount of infiltrated inflammatory cells were leaked from the pulmonary interstitial tissue. In conclusion, human umbilical cord mesenchymal stem cell transplantation for treatment of acute lung injury can reduce inflammatory factor levels and alleviate lung injury.