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。     

胎盘来源间充质干细胞研究进展

成人骨髓是临床应用时最常见的间充质干细胞(MSC)的来源。由于MSC在成人骨髓的数量以及分化能力随年龄的增长而减低,不同的胎儿组织被用来研究是否存在MSC。胎盘组织中MSC主要分布在羊膜和绒毛膜等来源于胎儿的组织,其抗原表达、分化潜能与骨髓间充质干细胞特征相似。     

人脐带血间充质干细胞的分离培养研究与展望

间充质干细胞(mesenchymal stemcells,MSCs)是一种多潜能干细胞。目前,MSCs主要取自骨髓,但由于骨髓源性MSCs存在高度病毒污染或其他污染的可能,且随着年龄的不断增长其细胞数量和扩增、分化能力出现明显下降趋势,因此,寻找一种能替代骨髓MSCs,并可弥补其缺陷的间充质干细胞来源,已越来越受到各国学者的关注。人脐带血(HUCB)是胎儿出生时脐带内及胎盘近胎儿一侧血管内的血液,含有丰富的干细胞和祖细胞,主要包含造血干细胞和MSCs。HUCB优势[1]在于:(1)脐血从分娩后的胎盘、脐带残端收集,其过程比从骨髓或胚胎获取干细胞简单。对于…     

大鼠脂肪和骨髓来源间充质干细胞的生物学特性比较

背景:除骨髓外,人们已从胎盘组织、脐带血肌肉组织、脂肪组织中分离到了间充质干细胞。目的:比较大鼠脂肪和骨髓间充质干细胞生物学特性和免疫调节功能的差异。方法:脂肪间充质干细胞和骨髓间充质干细胞分别来自BN大鼠的脂肪和骨髓。体外分离、纯化脂肪和骨髓来源间充质干细胞,进行细胞形态、表面标志、生长动力学、分化潜能鉴定;混合淋巴细胞反应比较两种细胞的免疫调节特性。结果与结论:脂肪间充质干细胞与骨髓间充质干细胞光镜和透射电镜下形态相似,第3代的脂肪间充质干细胞和骨髓间充质干细胞均高表达CD29,CD90,低表达CD34,CD45,CD11b;第3,4,5代脂肪间充质干细胞增殖速度明显快于骨髓间充质干细胞;两者都具有低免疫原性,可以抑制异基因抗原引起的T淋巴细胞增殖,且这种抑制作用与细胞数目成正相关,等量脂肪间充质干细胞和骨髓间充质干细胞抑制作用差异无显著性意义。结果证实脂肪间充质干细胞具有和骨髓间充质干细胞同样的低免疫原性和免疫调节功能。     

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     

间充质干细胞衰老的研究进展

<正>间充质干细胞(mesenchymal stem cells,MSCs)是一种具有高度自我更新能力、多向分化潜能和低免疫原性的异质细胞群,其来源广泛,可从骨髓、脐带、胎盘、脂肪、牙髓等多种组织中获得^[1]。MSCs可以在不同诱导条件下分化为多种组织和细胞,并分泌多种生物活性物质,其良好的免疫调节与组织修复功能为再生医学提供了新的策略及思路。     

胎盘间充质干细胞的体外诱导分化

背景：与骨髓、外周血、脂肪及胚胎等来源的间充质干细胞相比,脐带和胎盘组织来源的间充质干细胞具有来源广泛、易于获得、不引起供者不适以及不存在道德伦理争议等优势。 目的：探讨胎盘间充质干细胞体外分离、培养、传代的方法及其分化潜能,为胎盘间充质干细胞的临床应用提供基础研究依据。 方法：应用计算机检索2000年1月至2011年12月PubMed数据库、中国期刊全文数据库、万方数据库与胎盘间充质干细胞有关的文章,中文检索词为“胎盘间充质干细胞,体外,诱导分化”,英文检索词为“Placenta-derived mesenchymal stem cells,Induction and differentiation in vitro”。共检索到文献98篇,最终纳入符合标准的33篇文献。 结果与结论：目前国内胎盘间充质干细胞还处于基础研究阶段,国外胎盘间充质干细胞在实验研究方面已经取得了一定的进展,而且也在临床应用研究方面有了一定的突破。胎盘间充质干细胞具有来源广泛,取材方便,不引起供者不适以及不涉及伦理道德问题,与捐献骨髓或采集动员外周血相比,供者无痛苦,感染机会少, 扩增能力强等,在临床治疗方面有待于进一步深入研究。     

人骨髓间充质干细胞基因表达谱的芯片分析

目的：探讨骨髓间充质干细胞（MSCs）作为组织工程种子细胞的分子遗传学基础和临床应用价值。方法：通过密度梯度离心法和贴壁筛选法的联合应用，分离人骨髓间充质干细胞，流式细胞仪分析细胞同源性，UNIQ-10柱式Trizol总RNA抽提法获取足量的RNA，用于MSCs基因表达谱的分析研究。结果：流式细胞仪检测表明了所分离hMSCs的高度同源性，基因表达谱芯片分析首次揭示了MSCs表达多种间充质细胞和非间充质细胞的特异性mRNA，如脂肪细胞、成软骨细胞、成骨细胞、成肌细胞、造血支持基质、神经元、神经胶质细胞和上皮／内皮细胞等。结论：hMSCs具有高度可塑性和广泛分化潜能，是以干细胞工程为代表的现代组织工程的重要细胞来源。     

人胎盘间充质干细胞的超微结构及其吞噬功能

背景：胎盘间充质干细胞组织来源丰富,与骨髓间充质干细胞有着类似的形态、表面标记物、潜在分化能力,是人体理想的间充质干细胞来源之一。目前对胎盘间充质干细胞超微结构和吞噬功能的研究较少,对其在胎盘组织中的生理功能也鲜有探讨。目的：了解胎盘间充质干细胞的超微结构和吞噬功能。方法：分离培养5个正常娩出胎盘来源的胎盘间充质干细胞,经体外培养后在透射电子显微镜下观察细胞的超微结构;在培养上清中添加荧光微球,孵育3 h后用流式细胞仪检测细胞的吞噬能力。结果与结论：胎盘间充质干细胞在透射电镜下表现为细胞核比较大,核仁明显;核膜折叠、内陷显著,常染色质多,核浆比例正常。内质网系统发达,可见大量狭小或扩张的粗面内质网。胞浆内高尔基器、溶酶体常见;细胞表面微绒毛发达,偶可见细胞间连接。5个标本的胎盘间充质干细胞中均有部分细胞能吞噬荧光微球,最多有49.6%,最少有18.4%。上述胎盘间充质干细胞的超微结构特点提示细胞功能活跃,可合成和分泌大量蛋白质,并具有吞噬功能;而吞噬实验进一步验证胎盘间充质干细胞吞噬异物的能力,提示胎盘间充质干细胞在胎盘中可能起着维持微环境稳态、清除异物的功能。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：     

胎盘源间充质干细胞分离提取的新方法

背景：胎盘是获取间充质干细胞的理想来源,在干细胞治疗和再生医学中具有潜在广泛的用途,建立高效分离提取方法有助于其存储和临床应用。 目的：分析组织绞碎加酶液消化法培养人胎盘间充质干细胞的生物学特性。 方法：采用组织绞碎加酶液消化法分离、富集胎盘组织中间充质干细胞,应用流式细胞术检测细胞表面标记,MTT法分析增殖能力,并采用诱导剂定向诱导胎盘间充质干细胞成脂、成骨和成软骨分化。 结果与结论：胎盘组织来源间充质干细胞在体外可稳定长期传代培养,应用流式细胞仪检测结果显示 CD73、CD90、CD105表达阳性,CD11b、CD19、CD34、CD45、HLA-DR表达阴性。胎盘间充质干细胞增殖活跃,第3-5天进入对数生长期,第6天进入平台期。经诱导后,胎盘间充质干细胞具有向脂肪细胞、骨细胞、软骨细胞分化的潜能。结果表明联合利用组织绞碎和酶液消化法可高效获取胎盘间充质干细胞,在体外培养过程中生长稳定,增殖能力强,可长期传代。 1库：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：     

Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta

Recently we reported that second-trimester amniotic fluid (AF) is an abundant source of fetal mesenchymal stem cells (MSCs). In this study, we analyze the origin of these MSCs and the presence of MSCs in human-term AF. In addition, different parts of the human placenta were studied for the presence of either fetal or maternal MSCs. We compared the phenotype and growth characteristics of MSCs derived from AF and placenta. Cells from human second-trimester (mean gestational age, 19(+2) [standard deviation, +/- 1(+3)] weeks, n = 10) and term third-trimester (mean gestational age, 38(+4) [standard deviation, +/- 1] weeks, n = 10) AF, amnion, decidua basalis, and decidua parietalis were cultured in M199 medium supplemented with 10% fetal calf serum and endothelial cell growth factor. Cultured cells were immunophenotypically characterized, the adipogenic and osteogenic differentiation capacity was tested, and the growth kinetics were analyzed. The origin of fetal and maternal cells was determined by molecular human leukocyte antigen typing.We successfully isolated MSCs from second-trimester AF, amnion, and decidua basalis as well as term amnion, decidua parietalis, and decidua basalis. In contrast, MSCs were cultured from only 2 out of 10 term AF samples. The phenotype of MSCs cultured from different fetal and maternal parts of the placenta was comparable. Maternal MSCs from second-trimester and term decidua basalis and parietalis showed a significantly higher expansion capacity than that of MSCs from adult bone marrow (p < .05). Our results indicate that both fetal and maternal MSCs can be isolated from the human placenta. Amnion is a novel source of fetal MSCs, likely contributing to the presence of MSCs in AF. Decidua basalis and decidua parietalis are sources for maternal MSCs. The expansion potency from both fetal and maternal placenta-derived MSCs was higher compared with adult bone marrow-derived MSCs.     

Human Mesenchymal Stem Cells Derived from Bone Marrow Display a Better Chondrogenic Differentiation Compared with Other Sources

Mesenchymal stem cells (MSCs) are multipotent cells capable of differentiation into several mesodermal lineages. These cells have been isolated from various tissues, such as adult bone marrow, placenta, and fetal tissues. The comparative potential of these cells originating from different tissues to differentiate into the chondrogenic lineage is still not fully defined. The aim of our study was to investigate the chondrogenic potential of MSCs isolated from different sources. MSCs from fetal and adult tissues were phenotypically characterized and examined for their differentiation capacity, based on morphological criteria and expression of extracellular matrix components. Our results show that both fetal and adult MSCs have chondrogenic potential under appropriate conditions. The capacity of bone marrow-derived MSCs to differentiate into chondrocytes was reduced on passaging of cells. MSCs of bone marrow origin, either fetal or adult, exhibit a better chondrogenesis than fetal lung- and placenta-derived MSCs, as demonstrated by the appearance of typical morphological features of cartilage, the intensity of toluidine blue staining, and the expression of collagen type II, IX, and X after culture under chondrogenic conditions. As MSCs represent an attractive tool for cartilage tissue repair strategies, our data suggest that bone marrow should be considered the preferred MSC source for these therapeutic approaches.     

Human mesenchymal stem cells derived from bone marrow display a better chondrogenic differentiation compared with other sources

Mesenchymal stem cells (MSCs) are multipotent cells capable of differentiation into several mesodermal lineages. These cells have been isolated from various tissues, such as adult bone marrow, placenta, and fetal tissues. The comparative potential of these cells originating from different tissues to differentiate into the chondrogenic lineage is still not fully defined. The aim of our study was to investigate the chondrogenic potential of MSCs isolated from different sources. MSCs from fetal and adult tissues were phenotypically characterized and examined for their differentiation capacity, based on morphological criteria and expression of extracellular matrix components. Our results show that both fetal and adult MSCs have chondrogenic potential under appropriate conditions. The capacity of bone marrow-derived MSCs to differentiate into chondrocytes was reduced on passaging of cells. MSCs of bone marrow origin, either fetal or adult, exhibit a better chondrogenesis than fetal lung- and placenta-derived MSCs, as demonstrated by the appearance of typical morphological features of cartilage, the intensity of toluidine blue staining, and the expression of collagen type II, IX, and X after culture under chondrogenic conditions. As MSCs represent an attractive tool for cartilage tissue repair strategies, our data suggest that bone marrow should be considered the preferred MSC source for these therapeutic approaches.     

人脐静脉来源的间质干细胞体外分离扩增及多向分化的研究

目的： 探讨人脐静脉来源的间质干细胞(MSCs) 的体外分离、纯化、扩增和多向分化条件。 方法： 无菌条件下取正常人脐静脉,1%胶原酶Ⅱ消化脐静脉细胞，以IMDM作为培养基进行培养和纯化细胞，瑞氏染色和电镜观察形态；FACS检测其免疫表型和细胞周期；体外诱导成骨细胞、脂肪细胞分化，von Kossa染色、油红O染色和RT-PCR检测骨钙蛋白、脂蛋白脂酶mRNA的表达以检测细胞向成骨、成脂肪细胞分化情况。 结果： 脐静脉来源的细胞呈纤维样贴壁生长，瑞氏染色和电镜观察具有MSCs特征；FACS检测结果显示, 表达MSCs相关的抗原CD29、CD44、CD105，而CD31、CD13、CD34、CD45、HLA-DR为阴性；体外诱导成骨细胞、脂肪细胞分化成功。 结论： 人脐静脉来源的MSCs的细胞形态、生长特性、免疫表型、多向分化能力与骨髓来源的MSCs相似，可作为满足实验和临床需要的MSCs来源。     

人胎盘绒毛膜来源间充质干细胞的生物学特性

背景：骨髓是间充质干细胞的主要来源,但骨髓中间充质干细胞数量和质量会随着年龄的增长而逐渐降低。因此,寻找一种新的干细胞来源具有重要意义。 目的：分离研究了一种新的间充质干细胞来源——人绒毛膜来源间充质干细胞,并研究其生物学特性。 方法：将胎盘冲洗干净后,分离出脐带和羊膜组织,将剩余的胎盘组织进行原代和传代培养。通过短串联重复序列分析检测细胞是否来源于绒毛膜,用MTT法检测细胞生长方式,流式细胞仪分析细胞表型及干细胞标记,使用不同的诱导分化培养基检测其多向分化的能力。将该细胞与植物血凝素刺激的外周血单个核细胞共培养,用酶联免疫吸附试验检测上清γ-干扰素的水平。 结果与结论：短串联重复序列分析证明所得到的细胞来源于绒毛膜,这种绒毛膜来源的细胞生长呈典型的成纤维细胞形态。细胞表达常见的间充质干细胞表面标记CD90、CD73、CD105、CD44,不表达CD45,CD11b和CD34。同时,细胞也表达Nestin和Sox-2。在不同的条件培养基培养状态下,细胞可向成骨、成脂方向分化。这些结果证明所得到的细胞为人绒毛膜间充质干细胞。这些绒毛膜间充质干细胞能抑制植物血凝素刺激的人外周血单个核细胞分泌γ-干扰素。可见绒毛膜来源的间充质干细胞具有和传统的骨髓来源间充质干细胞具有相似的生物学特性。     

Searching for alternative sources of postnatal human mesenchymal stem cells: candidate MSC-like cells from umbilical cord

Mesenchymal stem cells (MSCs) have the capability for renewal and differentiation into various lineages of mesenchymal tissues. These features of MSCs attract a lot of attention from investigators in the context of cell-based therapies of several human diseases. Despite the fact that bone marrow represents the main available source of MSCs, the use of bone marrow-derived cells is not always acceptable due to the high degree of viral infection and the significant drop in cell number and proliferative/differentiation capacity with age. Thus, the search for possible alternative MSC sources remains to be validated. Umbilical cord blood is a rich source of hematopoietic stem/progenitor cells and does not contain mesenchymal progenitors. However, MSCs circulate in the blood of preterm fetuses and may be successfully isolated and expanded. Where these cells home at the end of gestation is not clear. In this investigation, we have made an attempt to isolate MSCs from the subendothelial layer of umbilical cord vein using two standard methodological approaches: the routine isolation of human umbilical vein endothelial cell protocol and culture of isolated cells under conditions appropriate for bone-marrow-derived MSCs. Our results suggest that cord vasculature contains a high number of MSC-like elements forming colonies of fibroblastoid cells that may be successfully expanded in culture. These MSC-like cells contain no endothelium- or leukocyte-specific antigens but express alpha-smooth muscle actin and several mesenchymal cell markers. Therefore, umbilical cord/placenta stroma could be regarded as an alternative source of MSCs for experimental and clinical needs.     

足月小样儿及正常胎儿脐血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脐血间充质干细胞(MSCs)的分离培养方法,观察其生物学特性,为脐血MSCs在临床的广泛应用奠定基础。方法无菌条件下采集足月小样儿和足月正常胎儿分娩脐血,密度梯度离心法分离脐血单个核细胞,采用MesencultTM培养基培养,观察脐血MSCs的生物学特性,免疫荧光方法检测其表面标记物的表达情况。结果采用MesencultTM培养基,正常儿脐血MSCs培养成功率为66.67%,相同培养条件下,足月小样儿脐血培养成功率低于正常体重胎儿的脐血。人脐血MSCs强表达CD29、CD44和CD90,不表达造血干细胞表面标志CD34。结论我们建立了最佳的脐血MSCs的分离培养方法。