间充质干细胞在肝脏疾病应用的研究现况

间充质干细胞（MSC）属于成体干细胞的一种,是一类具有自我更新和多向分化能力的多能干细胞。其来源丰富,免疫原性低,目前体内外实验均发现MSC可促进损伤肝脏修复,改善症状,提高存活率。通过调节肝脏局部和全身炎症反应和免疫紊乱发挥治疗作用。本文就MSC治疗肝脏疾病的研究现况进行综述。     

间充质干细胞在造血干细胞移植中的应用

间充质干细胞(MSCs)是一种具有自我更新和多向分化潜能的成体干细胞,存在于骨髓、脂肪组织、脐血及多种胎儿组织.它可分泌多种细胞因子及生长因子,促进造血干细胞(HSC)的增殖与分化.MSCs还具有免疫调节、抗炎和组织修复作用,可减轻移植物抗宿主病(GVHD)及其他移植相关并发症.     

骨髓间充质干细胞移植治疗终末期肝病:现状与前景

应用骨髓间充质干细胞及其分化来源的肝细胞移植来改善肝功能、桥接肝脏器官移植的治疗方法具有许多优势。目前已有实验研究和小规模临床实践运用这一方法,但是其临床疗效尚需评估,同时治疗机制也需要进一步探讨。该文就目前骨髓间充质干细胞分化为肝细胞及应用的临床和基础研究现状进行综述,对存在的问题及发展前景进行探讨。     

骨髓间充质干细胞抗肝纤维化的研究进展

肝纤维化及其终末病变肝硬化已严重危害全球人类健康,虽然慢性肝病的治疗手段和抗肝纤维化药物的研究已取得了很大进展,肝移植依然是最有效的治疗方案,但器官的紧缺却是一个现实问题。目前寻找有效的干预手段进行抗肝纤维化治疗已越来越受到大家的关注。近些年,大量基础及临床研究均证实在一定条件下利用骨髓间充质干细胞(MSCs)可以抑制肝星状细胞活化诱导其凋亡,实现肝纤维化逆转。随着干细胞技术的快速发展,基于骨髓间充质干细胞(MSCs)的细胞疗法在肝纤维化治疗领域的研究与应用已成为一个充满生命力的新方向。本文将对肝纤维化及基于MSCs的治疗机制进展及其应用进行综述。     

内源性骨髓间充质干细胞在肺纤维化中的研究进展

肺纤维化是多种病因引起的累及肺间质、肺泡、细支气管的肺部慢性、弥漫性、间质性肺疾病,尚无有效的治疗药物.目前,外源性骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BM-MSCs)移植作为一种新的干细胞疗法在治疗肺纤维化中的作用备受关注.目前对肺纤维化过程中内源性...     

骨髓间充质干细胞的研究进展

骨髓间充质干细胞是存在于骨髓中的具有高度自我更新能力和多向分化潜能的干细胞群体 ,具有支持造血、多向分化潜能以及在细胞和基因工程中具有潜在应用前景等特点 ,将在医学上具有重要的临床应用价值。     

骨髓间充质干细胞可塑性研究

骨髓间充质干细胞（bone marrow mesenchymal stem cells,MSCs）具有跨系统、甚至跨胚层分化的特性,称为MSCs的可塑性（plasticity）,成为细胞工程、再生医学中的主要选择细胞。但随着对成体干细胞可塑性质疑的出现,使MSCs是否具有转分化能力,存在着极大的分歧。对MSCs可塑性是否真正存在的进一步明确,越加显得急切和重要,也对干细胞基础理论及其临床应用具有指导性意义。     

骨髓间充质干细胞及其临床应用研究进展

骨髓间充质干细胞因具有容易获得、容易体外培养增殖、长期培养的过程中始终保持多向分化的潜能、抗原性小、组织修复能力强等特征,使之成为干细胞研究领域的热点和前沿,并被认为是最有前途的组织工程种子细胞之一。以干细胞工程为代表的现代组织工程学为组织器官的修复与替代提供了一个崭新的领域,并将此领域扩展到细胞替代治疗、支持造血、基因治疗等更多方面。     

骨髓间充质干细胞在大鼠实验中的移植途径及比较

骨髓间充质干细胞是具有自我更新能力和分化潜能的一类成体干细胞,经过局部微环境的诱导,可在体内外进行扩展,到晚期可分化成为多种细胞系。当组织受损伤时,可迅速到达损伤部位,分化为特异的组织细胞,参与组织修复。骨髓间充质干细胞这种惊人的分化及组织修复能力,为治疗退行性疾病和器官损伤性疾病提供广阔前景,故成为科研热点。国内外相关实验研究多以大鼠为动物模型,而骨髓间充质干细胞如何进入大鼠体内并定植,是实验成功的重要前提。因此如何找到最合适、最安全的移植途径将骨髓间充质干细胞有效地移植进入大鼠疾病模型体内的受损区域,是研究者关心的重点。本文就目前骨髓间充质干细胞在大鼠实验中不同移植途径进行综述,并比较各种途径的优缺点,希望能对临床科研工作提供参考,并期待能有更成熟的移植手段来推动骨髓间充质干细胞实验研究的进展。     

骨髓间充质干细胞在组织损伤局部微环境中的调节作用

骨髓间充质干细胞(mesenchymal stem cells,MSCs)是一种具有自我增殖和多向分化潜能的细胞,植入体内后对损伤组织具有一定的修复作用,研究发现MSCs在体内的分化效率极低(不足10%),故仅用其分化能力不能完全解释它良好的修复效能。新近研究表明,MSCs可通过旁分泌途径调节损伤局部的微环境,从而促进受损组织的修复,提示这种微环境的调节较其自身分化更具有临床意义。该文对MSCs在组织损伤局部微环境中的调节作用做一简要概述,为MSCs更广阔地应用于医学领域提供理论基础。     

移植途径对大鼠骨髓间充质干细胞归巢及肝功能恢复的影响

目的 探讨移植途径对骨髓间充质干细胞（MSCs）归巢及促进肝切除大鼠肝再生的影响.方法 建立肝切除大鼠模型,随机分为3 组,即肝切除对照组、尾静脉移植组和门静脉移植组.移植组分别经尾静脉和门静脉注射DAPI 标记的MSCs 约1.5×106/ 只,分别于第3 天和第9 天后采血清检测肝功能,第9 天处死大鼠取肝脏标本,并通过荧光显微镜观察两种移植途径对MSCs 向肝脏迁移的影响.结果 门静脉移植组（18.1 ± 3.4）个细胞/100 倍视野到肝脏归巢及定植的 MSCs 多于尾静脉移植组（7.6 ± 2.0）个细胞/100 倍视野,差异有统计学意义（P 〈 0.01）.术后第9 天各组大鼠肝功能均有好转,丙氨酸氨基转移酶（ALT）及天冬氨酸氨基转移酶（AST）3 组之间对比差异无统计学意义（F = 2.822,1.046,P = 0.057,0.365,P 〉 0.05）;但两移植组与单纯肝切除组比较血浆白蛋白（ALB）均有明显升高,差异具有统计学意义（F = 6.259,P = 0.006）;尾静脉移植组与门静脉移植组两移植组之间相比,差异无统计学意义（P 〉 0.05）.结论 移植途径对 MSCs 归巢、定植到肝脏有一定影响,门静脉途径优于外周静脉,MSCs 移植对肝大部切除大鼠肝功能恢复具有促进作用.     

Timing of transplantation of autologous bone marrow derived mesenchymal stem cells for treating myocardial infarction

It is still unclear whether the timing of intracoronary stem cell therapy affects the therapeutic response in patients with myocardial infarction.The natural course of healing the infarction and the presence of putative homing signals within the damaged myocardium appear to favor cell engraftment during the transendothelial passage in the early days after reperfusion.However,the adverse inflammatory environment,with its high oxidative stress,might be deleterious if cells are administered too early after reperfusion.Here we highlight several aspects of the timing of intracoronary stem cell therapy.Our results showed that transplantation of bone marrow mesenchymal stem cells at 2 4 weeks after myocardial infarction is more favorable for reduction of the scar area,inhibition of left ventricular remodeling,and recovery of heart function.Coronary injection of autologous bone marrow mesenchymal stem cells at 2 4 weeks after acute myocardial infarction is safe and does not increase the incidence of complications.     

骨髓间充质干细胞抑制小胶质细胞介导的炎症反应

目的研究骨髓间充质干细胞(Bone Marrow Mesenchymal Stem Cells,BMMSCs)对小胶质细胞介导的炎症反应的抑制作用。方法实验分为四组:组一:小胶质细胞(BV2)生长于DMEM(High Glucose)培养液中;组二:BV2细胞生长于加入脂多糖(LPS)的上述培养液中;组三:BV2细胞、BMMSCs共培养于加入LPS的上述培养液中;组四:骨髓间充质干细胞(BMMSCs)生长于加入LPS的上述培养液中。观察BV2细胞的生长状态、电镜超微结构变化及其分泌的炎症因子TNF-α表达量的变化。结果光镜下BV2细胞密度依次为:组一组三组二,组四中BMMSCs生长状态良好;电镜下可见组二BV2细胞内出现大量肿胀及空泡化的线粒体、内质网等细胞器,少见生长活跃多核仁细胞,同时可见大量崩解细胞,组三细胞状态明显好于组二;BV2细胞分泌的炎症因子TNF-α表达量依次为组二组三组一组四。结论 BM-MSCs抑制小胶质细胞介导的炎症反应,进而发挥神经保护作用。     

骨髓间充质干细胞向神经细胞分化的研究

无论是在体外实验、还是在体内实验,MSCs都可以向中枢神经系统(CNS)神经细胞分化,但争议颇多。因为功能性神经元不仅要具有典型神经元的形态、特异性标记,还要求具有可兴奋性、能和其他神经元形成突触联系、产生突触电位等,所以对于骨髓间充质干细胞是否能诱导出真正具有功能的神经元存在很大分歧。在此对MSCs向神经细胞诱导分化研究的现况、存在的问题及发展前景给以综述。     

供体骨髓干细胞输注联合肝移植治疗终末期肝病的近期效果观察

目的评价供体骨髓干细胞输注联合肝移植治疗终末期肝病的近期效果。方法2008年3月至2009年1月本中心30例肝移植受者分为两组,实验组8例,行同期供体骨髓干细胞联合肝脏移植;对照组22例,仅行肝脏移植,不行骨髓干细胞输注。观察两组受者术后免疫抑制剂（甲基强的松龙、他克莫司）用量、肝功能（谷氨酸转氨酶、天冬氨酸转氨酶等）变化、急性排斥反应和感染并发症发生情况,以及术后住院时间和费用。采用,检验、方差分析及确切概率法进行统计学分析。结果实验组术后甲基强的松龙总用量和出院时每日他克莫司用量显著低于对照组[甲基强的松龙分别为（1314±105）mg,（1884±256）mg,t=6．060,P=0．000;他克莫司分别为（3．73±0．35）mg／d,（4．93±0．62）mg／d,,=5．147,P=0．000]。术后第1天实验组血清谷氨酸转氨酶、天冬氨酸转氨酶均显著低于对照组[谷氨酸转氨酶分别为（875．2±325．5）IU／L,（1350．4±482．7）IU／L,t=2．543,P=0．016,天冬氨酸转氨酶分别为（646．2±184．9）IU／L,（1021．8±325．4）IU／L,t=3．067,P=0．005]。术后第3天也低于对照组[谷氨酸转氨酶分别为（252．9±35．8）IU／L,（343．5±47．8）IU／L,f=4．866,P=0．000,天冬氨酸转氨酶分别为（227．8±38．0）IU／L,（310．8±61．7）IU／L,t=3．545,P=0．001]。结论同期供体骨髓干细胞联合肝脏移植可以降低术后免疫抑制剂用量,减轻术后早期肝脏损伤,不增加治疗风险,是一种安全有效的治疗方法。     

人骨髓间充质干细胞脑内移植对食蟹猴脑出血的治疗作用

目的评价人骨髓间充质干细胞脑内移植对食蟹猴脑出血模型的治疗作用。方法符合普通级标准的成年食蟹猴12只,用自体股动脉抗凝血脑内注射方法建模后1周,用脑立体定位法在血肿周围植入人骨髓间充质干细胞,细胞数分别为高剂量5×106、低剂量1×106、对照组等体积生理盐水。利用MRI、PET、神经功能缺损评分和组织病理学对干细胞移植效果进行评价。结果神经功能评分显示干细胞移植1周后动物神经功能明显改善。PET结果显示干细胞移植后2周高剂量组血肿周围皮层、基底节核团的SUV%值与对照组间存在显著性差异(P=0.02)。移植后3周高、低剂量组血肿周围皮层、基底节核团的SUV%值与对照组间差异存在显著性(P值分别为0.03和0.04)。MRI显示剂量组血肿吸收速度大于对照组。病理检查可见剂量组坏死灶面积小于对照组,出血灶周围有大量新生血管生成,剂量组与对照组间差异存在显著性(P<0.01)。结论在损伤脑组织周围移植hBMSC可促进食蟹猴损伤神经组织的恢复,为hBMSC治疗脑出血的临床应用提供了重要实验依据。     

Isolation and characterisation of mesenchymal stem cells from adult mouse bone marrow

The future use of adult mesenchymal stem cells (MSCs) for human therapies depends on the establishment of preclinical studies with other mammals such as mouse. Surprisingly, purification and characterisation of murine MSCs were only poorly documented. The aim of this study was to purify mouse MSCs from adult bone marrow and to functionally characterise their abilities to differentiate along diverse lineages. Adherent cells from adult C57Bl/6J mouse bone marrow were depleted of granulo-monocytic cells and subsequently allowed to grow on fibronectin-coated dishes in presence of fetal bovine serum and growth factors. The growing fibroblastoid cell population primarily consisted of spindle- and star-shaped cells with significant renewal capacity as they were cultured until 30 passages (about 60 doubling population). We fully demonstrated the MSC phenotype of these cells by inducing them to differentiate along osteoblastic, adipocytic, and chondrocytic pathways. Mouse MSCs (mMSCs) sharing the same morphological and functional characteristics as human MSCs can be successfully isolated from adult bone marrow without previous mouse or bone marrow treatment. Therefore, mMSCs will be an important tool to study the in vivo behaviour and fate of this cell type after grafting in mouse pathology models.     

人骨髓间充质干细胞在精原细胞培养条件下生物学特征的观察

目的体外分离培养流产4h、八月龄男胎的骨髓间充质干细胞(BMSCs),并观察BMSC在精原细胞培养条件下的生物学特征,探讨人骨髓间充质干细胞诱导分化为精原细胞的可行性,为BMSCs作为"种子细胞"开辟新的途径。方法分离培养、鉴定人BMSCs,研究其在精原细胞培养条件下诱导培养BMSCs并观察其生物学特征。结果 BMSCs具有间充质细胞的特征;实验组细胞形态发生变化,免疫组化显示Oct-4和C-kit染色阳性;对照组细胞则无。结论人胎儿BMSCs有较强的增殖能力;人胎儿BMSCs在精原细胞培养条件下诱导培养,能表现出精原细胞的一些生物学特征。     

Evaluation of isolation methods and culture conditions for rat bone marrow mesenchymal stem cells

Bone marrow mesenchymal stem cells (bMSCs) are multipotent and preferred for cell therapy. However, the content of bMSCs is very low. To propagate a large number of primary bMSCs rapidly has become a prerequisite for bMSC study and application. Different methods of isolating and culturing bMSC were used and compared among groups: bMSCs of group A are isolated using direct adherence method and cultured by conventional medium changing; of group B are isolated using direct adherence method and cultured by low volume medium changing; of group C are isolated using density gradient centrifugation and cultured by conventional medium changing; of group D are isolated using density gradient centrifugation and cultured by low volume medium changing. The average population doubling time (PDT), average generation time and the cumulative cell doubling level were calculated for every group. bMSCs cultured with complete medium containing 10, 11 and 15 % FBS were allocated into group a, b and c separatedly. Cell numbers were counted everyday under a microscope, the population doubling level curve was plotted and PDT was calculated. The growth curve of bMSC in group a, b and c was made. Both density gradient centrifugation and direct adherence methods obtained relatively pure bMSCs. A larger quantity of primary bMSCs were obtained by direct adherence. bMSC proliferation was faster when cultured via the low volume medium changing method at a serum concentration of 11 % than the other methods. Isolating bMSC by direct adherence and culturing by low volume medium changing at a serum concentration of 11 % is preferential for bMSC propagation.     

Vitreous cryopreservation of tissue engineered bone composed of bone marrow mesenchymal stem cells and partially demineralized bone matrix

Cryopreservation of tissue engineered products by maintaining their structure and function is a prerequisite for large-scale clinical applications. In this study, we examined the feasibility of cryopreservation of tissue engineered bone (TEB) composed of osteo-induced canine bone marrow mesenchymal stem cells (cBMSCs) and partially demineralized bone matrix (pDBM) scaffold by vitrification. A novel vitreous solution named as VS442 containing 40% dimethyl-sulfoxide (DMSO), 40% EuroCollins (EC) solution and 20% basic culture medium (BCM) was developed. After being cultured in vitro for 8 days, cell/scaffold complex in VS442 was subjected to vitreous preservation for 7 days and 3 months, respectively. Cell viability, proliferation and osteogenic differentiation of cBMSCs in TEB after vitreous cryopreservation were examined with parallel comparisons being made with those cryopreserved in VS55 vitreous solution. Compared with that cryopreserved in VS55, cell viability and subsequent proliferative ability of TEB in VS442 after being rewarmed were significantly higher as detected by live/dead staining and DNA assay. The level of alkaline phosphatase (ALP) expression and osteocalcin (OCN) deposition in VS442 preserved TEB was also higher than those in the VS55 group since 3 days post-rewarm. Both cell viability and osteogenic capability of the VS55 group were found to be declined to a negligible level within 15 days post-rewarm. Furthermore, it was observed that extending the preservation of TEB in VS442 to 3 months did not render any significant effect on its survival and osteogenic potential. Thus, the newly developed VS442 vitreous solution was demonstrated to be more efficient in maintaining cellular viability and osteogenic function for vitreous cryopreservation of TEB over VS55.