胎盘源间充质干细胞对T淋巴细胞体外增殖的影响

目的从人胎盘组织中分离培养获得间充质干细胞（mesenchymal stem cells,MSCs）,并进一步研究其对T淋巴细胞体外增殖的影响。方法首先从胎盘组织中分离培养胎盘间充质干细胞（PMSCs）,在体外观测其形态,并通过细胞表面抗原表达、分化潜能等特征进行鉴定;然后将体外分离培养扩增的PMSCs,按照不同比例加入双向混合淋巴细胞培养体系（mixed lymphocyte reaction,MLR）中,共同培养6d后,测定T淋巴细胞的增殖。结果从人胎盘组织中分离培养获得问充质干细胞,具有与骨髓间充质于细胞（BMSCs）极为相似的细胞形态及细胞表面标志,表达CD29、CD44和CD105,不表达CD34、CD45、CD106和HLA—DR,并且还具有与BMSCs相似的跨胚层分化能力,能在一定条件下被诱导分化出神经元样细胞。PMSCs与同种异体混合淋巴细胞共同培养能抑制T淋巴细胞的体外增殖。结论胎盘组织是MSCs的有效来源,PMSCs具有与BMSCs相似的生物学特性及免疫调节机制。它为问充质干细胞的研究提供了又一重要的细胞来源。     

人骨髓间充质干细胞的分离、培养与鉴定

目的:探讨密度梯度离心结合贴壁筛选法分离培养人骨髓间充质干细胞(MSCs)的条件,为组织工程选择合适的种子细胞奠定基础.方法:采用密度梯度离心法将人MSCs自少量骨髓中分离并培养,观察其增殖和生长特性,绘制生长曲线,测定贴壁率,免疫组化与流式细胞仪细胞表型鉴定.结果:人MSCs生物性状稳定.不同代次生长曲线相似,第5～6d细胞数日最多;传代后10 h贴壁率高达90%.MSCs表达CD44、CD90,但不表达CD34、CD45.结论:密度梯度离心法是分离人MSCs的理想方法.MSCs能为组织工程提供足量种子细胞.     

人骨髓源和胎盘源间充质干细胞对T细胞增殖抑制作用的比较

目的:比较研究人骨髓源和胎盘源间充质干细胞介导的T细胞增殖抑制作用机制。方法:应用流式细胞术(FCM)分别检测B7H4和PDL1在人骨髓源间充质干细胞(HBMSCs)和胎盘源间充质干细胞(HPMSCs)上的表达;应用抗体阻断试验分析B7H4、PDL1分别在HBMSCs和HPMSCs对T细胞增殖及周期影响中的作用。结果:HBMSCs上高表达免疫负性调控分子B7H4,而HPMSCs上高表达免疫负性调控分子PDL1。分别应用B7H4mAb和PDL1mAb阻断,可使HBMSCs和HPMSCs对PHA激发的T细胞增殖抑制作用明显减弱;下调T细胞周期中G0/G1期细胞数量,上调S期细胞数量,明显减弱HBMSCs和HPMSCs对T细胞周期的影响。结论:HBMSCs和HPMSCs可通过表达不同的免疫负性调控分子介导T细胞的增殖抑制作用。     

人骨髓间充质干细胞的生物学特性

目的建立人胚骨髓间充质干细胞(MSCs)分离及培养的方法,探讨体外培养MSCs的生物学特性.方法通过密度梯度离心、贴壁法分离培养人胚MSCs,在倒置显微镜下连续观察细胞的形态变化.应用流式细胞术测定细胞周期和CD44、CD34、CD45表达,并研究其增殖及生长特征.结果原代及传代培养显示,14代以前的MSCs具有活跃的增殖能力,细胞周期分析显示有82%的MSCs处于G0/G1期.MSCs阳性表达CD44,阴性表达CD34、CD45.结论体外培养14代以前的的MSCs生长稳定,增殖较快,可作为组织工程的种子细胞.     

C57小鼠脂肪及骨髓间充质干细胞生物学特性的比较

背景：研究发现,脂肪源干细胞具有和骨髓间充质干细胞一样的贴壁和形成成纤维样克隆特性,并具有向骨、脂肪、软骨等多系分化的能力。 目的：比较C57小鼠脂肪源间充质干细胞和骨髓间充质干细胞的生物学特点。 方法：在无菌的条件下分别从C57小鼠的脂肪和骨髓中获取脂肪间充质干细胞和骨髓间充质干细胞。体外分离、培养并将脂肪间充质干细胞和骨髓间充质干细胞传至第3代,进行细胞形态、表面标记、生长动力学分化潜能测定和Notch信号相关基因的检测。 结果与结论：脂肪间充质干细胞和骨髓间充质干细胞形态学相似,第3代的脂肪间充质干细胞和骨髓间充质干细胞均表达CD29、CD105、Sca-1,不表达CD34、CD133,但骨髓间充质干细胞还表达CD45;生长曲线和细胞克隆分析显示脂肪间充质干细胞的增殖速度明显比骨髓间充质干细胞快;脂肪间充质干细胞和骨髓间充质干细胞均可向成骨、成脂、成软骨诱导分化,脂肪间充质干细胞更易向成骨诱导;Notch相关基因检测显示脂肪源干细胞的Jagged-1表达水平明显比骨髓间充质干细胞低,而Hes-1的表达水平脂肪源干细胞明显高于骨髓间充质干细胞的表达水平。提示脂肪间充质干细胞比骨髓间充质干细胞扩增能力更强,更易向成骨分化,可能与Hes-1表达水平有关。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：     

人骨髓间充质干细胞的生物学特性

目的 建立人胚骨髓间充质干细胞(MSCs)分离及培养的方法，探讨体外培养MSCs的生物学特性．方法 通过密度梯度离心、贴壁法分离培养人胚MSCs，在倒置显微镜下连续观察细胞的形态变化．应用流式细胞术测定细胞周期和CD44、CD34、CD45表达，并研究其增殖及生长特征．结果 原代及传代培养显示，14代以前的MSCs具有活跃的增殖能力，细胞周期分析显示有82％的MSCs处于Gp／Gl期．MSCs阳性表达CD44，阴性表达CD34、CD45．结论 体外培养14代以前的的MSCs生长稳定，增殖较快，可作为组织工程的种子细胞．     

小鼠骨髓间充质干细胞的分离培养及形态学观察

目的建立一种简单实用的小鼠骨髓间充质干细胞(MSCs)的分离培养方法,通过了解其细胞生物学特性,为MSCs的应用提供实验依据。方法采用差速贴壁法体外分离、扩增MSCs,倒置显微镜观察原代及传代细胞的形态及生长过程。结果在小鼠骨髓间充质干细胞的培养过程中,血液系细胞在换液过程被去除,成纤维细胞污染经差速贴壁法也可去除。获得的骨髓间充质细胞形态较均一,生长状态良好。结论采用差速贴壁培养法可获得一定纯度的MSCs,此法简单、实用,并且获得的细胞生长状态良好,增殖能力强.     

hCMSC体外培养的优化及生物学特性的初步分析

目的:探索从人胎盘绒毛组织体外分离培养获得间充质干细胞(hCMSC)和hCMSC体外有效扩增的方法,并对hCMSC生物学特性作初步分析.方法:免疫组织化学染色确定胎盘绒毛组织中间充质干细胞的组织分布,选取相应的绒毛组织和采用Ⅳ型胶原酶消化法,获得细胞悬液,进行原代传代培养;流式细胞术检测细胞表面标志,采用细胞计数和MTT法分析细胞的增殖;叔丁对甲氧酚(Butylated hydroxyanisole,BHA)和DMSO联合诱导hCMSC向神经元样细胞分化;RT-PCR分析flt3基因表达.结果:免疫组织化学染色结果显示,胎盘绒毛中轴血管周围富有CD105、CD90阳性的间充质干细胞,采用机械分离和Ⅳ型酶消化法获取绒毛层富含hCMSC的组织细胞;bFGF是良好的hCMSC体外增殖因子,而FL对人胎盘绒毛层间充质干细胞具有更明显的促增殖作用;体外扩增的hCMSC高表达CD73、CD90、CD105,低表达CD14、CD34、CD45、HLA-DR,以及免疫负性调控分子PDL-1呈现表达,并且具有向神经元样细胞分化的潜能.结论:胎盘绒毛组织中存在低免疫原性的间充质干细胞,bFGF和FL(flt3 ligand)能有效促进胎盘绒毛层间充质干细胞体外增殖且保持其分化潜能.     

人脐血源性MSCs的免疫调节作用

目的 从人脐血中分离、培养间充质干细胞(MSCs)并探讨其对淋巴细胞的免疫调节作用.方法 淋巴细胞分离液分离人脐血单个核细胞,利用贴壁筛选法通过多次传代得到MSCs,流式细胞仪测定细胞表型;将获得的MSCs分别以不同数量加入到外周血混合淋巴细胞培养体系和植物血凝素(PHA)刺激的外周血淋巴细胞转化体系中,用H3-TdR标记β液体闪烁计数仪检测细胞增殖,观察脐血来源的MSCs对混合淋巴细胞反应和淋巴细胞转化的影响.结果 从人脐血中分离获得的贴壁细胞,呈成纤维样的细胞形态,CD29、CD105和CD166表达阳性,CD14、CD34和CD45表达阴性;人脐血源性MSCs在体外对混合淋巴细胞反应和PHA诱导的淋巴细胞转化均具有明显的抑制作用,抑制作用与细胞数量呈正相关.结论 从人脐血中可以成功地分离出MSCs,其对同种异体淋巴细胞具有免疫调节作用,为其作为骨组织工程异基因种子细胞来源打下基础.     

人脐带间充质干细胞的分离培养及生物学特性

目的建立一种稳定、高效的分离培养人脐带间充质干细胞(MSCs)的方法,并对其生物学特性进行鉴定。方法分别采用组织块贴壁法和双酶消化法分离培养人脐带MSCs,对比其培养成功率;流式细胞术检测脐带MSCs表面抗原及细胞周期;采用丹参联合生长因子的方法诱导其向神经细胞分化,观察脐带MSCs向神经元分化潜能。结果组织块贴壁法获得脐带MSCs成功率高;细胞表达CD29、CD44和CD90,不表达造血干细胞表型CD34。细胞倍增时间为30h,G0-GI期和S+G2+M期所占比例分别为78.47%和21.53%。脐带MSCs在体外经诱导分化出现神经元样细胞改变,且表达神经元特异性烯醇化酶(NSE)。结论成功建立了高效稳定的脐带MSCs分离培养方法,脐带MSCs具备较强的分化潜能和增殖能力,为临床移植治疗提供了理想的细胞来源。     

骨髓间充质干细胞研究进展

骨髓间充质干细胞是干细胞领域的研究热点之一.虽然近几年来有关间充质干细胞的研究已取得了很大进展,但仍有很多问题有待进一步解决.主要对间充质干细胞的异质性、分化与融合的问题、修复组织的机制、免疫性及研究方法等问题进行了综述.     

Aging of mesenchymal stem cells

The role of adult mesenchymal stem cells (MSC) in tissue maintenance and regeneration has received significant attention of late. Questions arise to what extent these cells are either subject to, or causes of aging; whether age-related changes in these cells are due to intrinsic factors or induced by the somatic environment. This review collates and examines recent data in support of these different theories. By means of introduction, a brief overview is given of current MSC definitions and their basic role in tissue regeneration followed by a comparative analysis of gerontological studies involving MSC. Evidence for extrinsic aging and various aging markers relating to morphology, proliferation, signalling, senescence markers, telomeres and telomerase, and other indicators is discussed. We observe that while the literature might often appear to conflict, many apparent discrepancies are attributable to inconsistent methods of extracting and isolating MSC which in fact contains various subsets of adult stem cells, varying not only in their differentiation potential but also in their vulnerability to senescence--ranging from quasi-somatic lifespan to perennial vigour. Thus, mesenchymal stem cells emerge as both subject to and key mediators of organismal aging.     

骨髓间充质干细胞研究进展

骨髓间充质干细胞是干细胞领域的研究热点之一。虽然近几年来有关间充质干细胞的研究已取得了很大进展,但仍有很多问题有待进一步解决。主要对间充质干细胞的异质性、分化与融合的问题、修复组织的机制、免疫性及研究方法等问题进行了综述。     

High-potential human mesenchymal stem cells

Bone marrow-derived stromal mesenchymal stem cells (MSCs) have been characterized in vitro by their growth characteristics, the expression of a panel of surface antigens, and their potential to differentiate into mesenchymal lineages. They can be separated by physical methods as well as by immunological or chemical separation or cultivation. Different protocols are used in different laboratories, making the comparison of various reported MSC populations difficult. Here we describe a population of bone marrow-derived adult stem cells that has been separated on a Percoll gradient with low density. It is characterized by an extraordinary high proliferative potential and a conserved phenotype characteristic of MSCs that retain their plutipotentiality in culture, as evidenced by their ability to differentiate into osteo-, chondro-, and adipogenic lineages. Separation of these cells provide an effective and convenient method for rapid expansion of pluripotential human MSCs for clinical use where large amounts of stem cells are needed.     

间充质干细胞的免疫调节作用

间充质干细胞(MSC)已成功从动物体尤其骨髓中分离获得.干细胞,具有向多种组织和细胞如骨、脂肪、成软骨细胞等分化的潜能,这种潜能越来越引起人们关注,它可向受损的组织定向迁移以修复受损组织.体外资料研究表明,不管协同刺激信号存在与否,都具有低免疫原性.在体外MSC还具有免疫抑制活性,可抑制同种异体抗原或丝裂霉素刺激的T细胞扩增,阻止细胞毒性T细胞增殖,同时还具有调节炎性细胞的免疫活性使其向抗炎细胞分化的作用.因此,系统了解.MSC的生物学、免疫学特性将为人们深入阐明其免疫调节机制,并将其用于治疗自身免疫性病、组织修复及移植提供依据.     

骨髓基质干细胞的研究进展

骨髓基质干细胞具有取材简单、增殖速度快、抗原性小、培养过程中始终保持多向分化的潜能等特点,已经成为干细胞研究领域的热点,是最好的组织工程种子细胞之一。骨髓基质干细胞研究为组织器官的修复和替代提供了一个新的选择,具有广阔的临床应用前景。     

Proteomics of primary mesenchymal stem cells

Tissue and functional regeneration takes place in the body at various stages throughout life. However, bone, cartilage, tendons, blood vessels and cardiac muscle have a limited capacity for self repair and, after injury or disease, the regenerative ability of these adult tissues is often insufficient and leads to nonfunctional scar tissue. In this context, mesenchymal stem cells, which are adult multipotential progenitors of mesoderm cells (osteoblasts, chondrocytes, adipocytes and stroma cells), represent a major hope for tissue-engineered replacement and regenerative medicine. Furthermore, the autologous use of these cells prevents immunological responses against new tissues and the risks of disease transmission from donors, which are both common problems of organ transplantation. While the existence of mesenchymal stem cells is undisputed, many questions remain regarding their self-renewal and capacity to differentiate, their homogenous nature as a cell population throughout the body and their true potential in regenerative medicine. In this article, the proteomics studies carried out to characterize mesenchymal stem cells and to help understand their physiology are reviewed.     

胎盘间充质干细胞的应用研究

背景：胎盘间充质干细胞因其具有来源广泛、免疫原性低、不涉及伦理问题等优点成为种子细胞的新来源。 目的：阐述胎盘间充质干细胞的来源、生物学特性及应用最新研究进展。 方法：检索 PubMed、ScienceDirect、OvidSP、CNKI 数据库相关文章,检索时限为2003至 2015年,英文检索词为“Placenta,Mesenchymal stem cells,The placenta mesenchymal stem cells, Cell transplantation , Application mechanism”,中文检索词为“胎盘,间充质干细胞,胎盘间充质干细胞,细胞移植,应用机制”,从中筛选出与主题相关且论据可靠的部分文献,最终纳入57篇文章进行归纳综述。 结果与结论：目前已成功分离培养出胎盘间充质干细胞,并对其生物学特性进行研究,证明其具有干细胞源性及多向分化潜能。目前有较多关于胎盘间充质干细胞应用于实验动物及临床的研究,在骨组织工程、血管再生及神经组织等修复过程中均显示出了巨大的潜力,但胎盘间充质干细胞的具体应用机制目前尚不清楚,尚处于探索阶段,在胎盘间充质干细胞广泛应用于临床之前,仍有许多问题有待进一步研究明确,以确保其安全性和有效性。  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     

Immunoregulatory function of mesenchymal stem cells

Mesenchymal stem cells (MSC) are a rare subset of stem cells residing in the bone marrow where they closely interact with hematopoietic stem cells and support their growth and differentiation. MSC can differentiate into multiple mesenchymal and non-mesenchymal lineages, providing a promising tool for tissue repair. In addition, MSC suppress many T cell, B cell and NK cell functions and may affect also dendritic cell activities. Due to their limited immunogenicity, MSC are poorly recognized by HLA-incompatible hosts. Based on these unique properties, MSC are currently under investigation for their possible use to treat immuno-mediated diseases. However, both their condition of immunoprivilege and their immunosuppressive function have recently been challenged when analyzed under particular experimental conditions. Thus, it is likely that MSC effects on the immune system may be deeply influenced not only by cell-to-cell interactions, but also by environmental factors shaping their phenotype and functions.     

The concept of mesenchymal stem cells

In this chapter we examine whether criteria usually defining adult tissue stem cells apply to mesenchymal stem cells (MSCs) that give rise to cells of the skeletal connective tissues. MSCs appear to constitute a heterogeneous population of undifferentiated and committed, lineage-primed cells, capable of: homing upon engraftment to a number of growth microenvironments, extensive proliferation, producing large numbers of differentiated progeny, and functional tissue repair after injury. In addition, MSCs are extensively distributed throughout tissues, and bone marrow MSCs provide the stromal component of the niche of hematopoietic stem cells. The capacity of apparently differentiated mesenchymal cells to shift their differentiation pathway with changing microenvironmental conditions (known as differentiation plasticity) may be due to de-differentiation and reprogramming in MSCs. Because they present several features setting them apart from other stem cells, MSCs may constitute another paradigm for stem cell systems, where self-renewal and hierarchy are no longer essential, but where plasticity is the major characteristic.