A Novel Population of Mesenchymal Progenitors with Hematopoietic Potential Originated from CD14- Peripheral Blood Mononuclear Cells

Hemopoietic system derived progenitor cells with mesenchymal features have been identified including CD14⁺ monocyte-derived progenitors. However, it is unclear whether there are mesenchyme derived progenitors with hematopoietic potential. Herein, we identified a novel CD14^- cell-derived population with both mesenchymal and hematopoietic features in rat peripheral blood, and this cell population is different from the CD14⁺ monocyte-derived progenitors but designated peripheral blood multipotential mesenchymal progenitors (PBMMPs). Phenotype analysis demonstrated expression of mesenchymal markers in PBMMPs including BMPRs, Endoglin/CD105, Fibronectin (Fn), Vimentin (Vim), Collagen (Col) I/II/III along with hematopoietic marker CD34. CD14⁺ cell-derived population shared the same characteristics with CFs. In mixed culture of CD14⁺ and CD14^- cells, PBMMPs were a predominant component and expressed CD29^high, CD73^high, CD34^high, CD45^low and CD90. Except for the value of mixed T lymphocytes and CD14⁺ cell-derived population, hematopoietic characters of cultured PBMMPs were indicated by CD14^-/CD34⁺/CD45^-/CD90⁺. The mesenchymal origin was further confirmed by comparing PBMMPs with bone marrow stromal cells. Finally, we transplanted PBMMPs into a skin wound model, and results showed the specific potential of PBMMPs in not only extracellular matrix secretion but epidermal regeneration. This study provides evidence that peripheral blood contains common hematopoietic-mesenchymal progenitors from both hematopoietic and mesenchymal lineages, and CD34⁺ mesenchymal progenitors are a possible alternative source of epidermal cells in wound healing.     

3D co-culture of hematopoietic stem and progenitor cells and mesenchymal stem cells in collagen scaffolds as a model of the hematopoietic niche

Here, we propose a collagen-based three-dimensional (3D) environment for hematopoietic stem and progenitor cells (HPC) with mesenchymal stem cells (MSC) derived either from bone marrow (BM) or umbilical cord (UC), to recapitulate the main components of the BM niche. Mechanisms described for HPC homeostasis were systematically analyzed in comparison to the conventional liquid HPC culture. The 3D-cultivation allows dissecting two sub-populations of HPC: (I) HPC in suspension above the collagen gel and (II) migratory HPC in the collagen fibres of the collagen gel. The different sites represent distinct microenvironments with significant impact on HPC fate. HPC in niche I (suspension) are proliferative and a dynamic culture containing HPC (CD34⁺/CD38^-), maturing myeloid cells (CD38⁺, CD13⁺, CAE⁺) and natural killer (NK) cells (CD56⁺). In contrast, HPC in niche II showed clonal growth with significant high levels of the primitive CD34⁺/CD38^- phenotype with starting myeloid (CD13⁺, CAE⁺) differentiation, resembling the endosteal part of the BM niche. In contrast, UC-MSC are not adequate for HSC expansion as they significantly enhance HPC proliferation and lineage commitment. In conclusion, the 3D-culture system using collagen and BM-MSC enables HPC expansion and provides a potential platform to dissect regulatory mechanisms in hematopoiesis.     

<Emphasis Type="Italic">In vitro</Emphasis> comparison of immunological properties of cultured human mesenchymal cells from various sources

Cell-mediated cytotoxicity was studied in vitro during the interaction of bone marrow mesenchymal stem cells, fibroblast-like cells from newborn umbilical cord, and skin fibroblasts of an adult donor with peripheral blood mononuclear cells. Independently on the origin, mesenchymal cells were not lysed with allogeneic natural killer cells and cytotoxic lymphocytes. Mixed cultures of mesenchymal cells had no cytotoxic effect on peripheral blood mononuclear cells and did not activate proliferation of T and B lymphocytes, natural killer cells, and CD14⁺ lymphocytes. In vitro experiments showed that mesenchymal cells of different origin and allogeneic immunocompetent cells are tolerant to each other. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 2, pp. 188–191, February, 2008     

3种组织来源的间充质干细胞体外成骨能力的比较

目的比较成人骨髓间充质干细胞(BMSCs)、人脐带间充质干细胞(UC-MSCs)和人胎盘间充质干细胞(P-MSCs)的成骨能力。方法用含10%胎牛血清的DMEM/Ham's F-12培养液培养3种MSCs,CCK8法检测增殖能力,流式细胞仪鉴定3种细胞。碱性磷酸酶(ALP)和茜素红染色观察细胞经成骨诱导后成骨分化蛋白-ALP的分泌和矿化钙结节的沉积。实时荧光定量PCR(RT-q PCR)法检测MSCs骨再生相关基因的表达。Western blot方法检测MSCs成骨再生相关基因的蛋白表达。结果 MSCs在第3天进入对数增殖期。3种细胞的表面标志物阳性率:CD44、CD90和CD105均高于98%。3种MSCs成骨诱导9 d时,3种MSCs的实验组均表达大量成骨分化蛋白-ALP,成骨诱导18 d时3种MSCs均呈现较好的矿化能力;3种MSCs成骨诱导9 d时,实验组RUNX2和ALP基因显著性高表达(P0.05),成骨诱导18 d时,实验组RUNX2和骨钙素(OCN)亦显著性高表达(P0.05);3种MSCs成骨诱导9 d时,实验组均检测到RUNX2和ALP的蛋白表达;成骨诱导18 d时,实验组细胞亦检测到RUNX2和OCN的蛋白表达。结论 UC-MSCs和P-MSCs具有良好的成骨分化能力,有望作为骨组织工程的种子细胞用于治疗骨缺损。     

γ干扰素联合脂多糖诱导人脐带间充质干细胞向MSC2极化

文题释义： γ干扰素联合脂多糖模拟炎症微环境：间充质干细胞的免疫抑制能力不是内在的,而是由促炎细胞因子诱导的。间充质干细胞暴露于炎症信号可显著增强其对T细胞、单核细胞/巨噬细胞和树突状细胞的免疫抑制作用。γ干扰素由活化T细胞和NK细胞产生,激活抗原提呈细胞,上调转录因子T-bet 而促进Th1细胞分化;脂多糖是革兰阴性细菌细胞壁外壁的组成成分,是免疫反应的强烈刺激剂,具有通过信号转导途径促进各种炎性细胞因子分泌的作用。 间充质干细胞可极化为MSC1和MSC2两种类型：间充质干细胞既可以抑制免疫应答,也可以促进免疫应答,不同的炎症递质会诱导间充质干细胞极化分型并表现出截然相反的免疫调节作用,这一特性称为间充质干细胞免疫调节的可塑性。间充质干细胞可以通过下游Toll样受体信号极化成2种类型：MSC1和MSC2。文献报道Toll样受体4引发的MSC1主要发挥促炎功能,而Toll样受体3引发的MSC2主要发挥免疫抑制功能。 背景：间充质干细胞的免疫调节特性已被临床广泛应用于自身免疫性疾病和移植物抗宿主病,但是其免疫调节的可塑性导致间充质干细胞临床治疗效果出现异质性和不稳定性。 目的：探索γ干扰素联合脂多糖模拟炎症微环境诱导人脐带间充质干细胞向MSC2极化的作用。 方法：体外分离培养人脐带间充质干细胞,进行形态学、表面标志物、成脂及成骨诱导分化能力鉴定。分别用γ干扰素(10 μg/L)、脂多糖(100 μg/L)及二者联合刺激人脐带间充质干细胞24 h,与植物凝集素刺激的外周血单个核细胞直接共培养或Transwell间接共培养5 d。流式细胞术检测共培养不同时间点调节性T细胞和Th1细胞比例,荧光定量PCR检测人脐带间充质干细胞的Toll样受体2,3,4 mRNA表达水平。 结果与结论：①人脐带间充质干细胞呈梭形或成纤维形,高表达CD73、CD90、CD105,低表达CD34、CD45、HLA-DR;②在直接与间接共培养条件下,γ干扰素联合脂多糖刺激人脐带间充质干细胞均可增加调节性T细胞的比例,优于γ干扰素或脂多糖单独刺激组、未刺激组及外周血单个核细胞对照组(P < 0.05);③Th1细胞比例随共培养时间的延长呈逐渐下降的趋势;④在间接共培养的条件下,γ干扰素联合脂多糖刺激人脐带间充质干细胞更早向免疫抑制型MSC2极化,Toll样受体3表达显著增高(P < 0.05);⑤以上结果表明间接共培养体系下γ干扰素(10 μg/L)联合脂多糖(100 μg/L)是诱导人脐带间充质干细胞向MSC2极化的高效组合,并且MSC2的免疫抑制作用不依赖于细胞间的直接接触,为MSC2来源外泌体将来应用于临床研究奠定实验基础。 ORCID: 0000-0001-7335-2558(黄恬) 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     

Characteristics of bone marrow cells under conditions of impaired innervation in patients with spinal trauma

We studied quantitative and functional parameters of bone marrow stem cells and mature lymphocyte population under conditions of impaired innervation in patients with injuries to the cervical and thoracic portions of the spinal cord. Our findings indicated the absence of deficiency of quantitative and proliferative potentials of stem cells and demonstrated intact subpopulation structure of mature lymphocytes and T-cell proliferative activity similar to that in donors. The content of CD34⁺ cells in patients did not differ from that in donors. The percentage of CD34⁺CD38⁻ hemopoietic stem cells was elevated in patients, presumably due to increased proliferative activity of hemopoietic stem cells. The possibility of derivation and in vitro culturing of fibroblast-like cells with mesenchymal stem cell phenotype was demonstrated. __________ Translated from Kletochnye Tekhnologii v Biologii i Meditsine, No. 1, pp. 8–11, January, 2006     

采用组织贴壁法从整根脐带分离培养间充质干细胞及其生物学特性的研究

 为了建立从整根脐带中分离培养间充质干细胞(UC-MSCs)的技术,并对其生物学特性进行研究。采用组织贴壁法分离UC-MSCs, 并通过传代进行纯化和扩增培养, 绘制生长曲线：用流式细胞仪检测UC-MSCs表面抗原及细胞周期; 在特定诱导体系中,检测UC-MSCs向脂肪、成骨及软骨分化的能力;采用RT-PCR检测多能干细胞标志多能干细胞标志Oct4, Sox-2, Nanog mRNA水平。结果表明,　成功建立了UC-MSCs分离培养的方法;流式细胞仪检测结果显示, 贴壁细胞均表达CD73 、CD90 、CD105, 不表达造血细胞表型CD34、CD45 和HLA-DR ;细胞倍增时间为(24.04±0.49)h , 细胞周期分析表明,G0～G1 期和S+ G2 + M 期所占比例分别为81.56 %和18.44%;UC-MSCs能够向脂肪、成骨和软骨分化;表达Oct4, Sox-2, Nanog基因。结论：组织贴壁法是一种较好的分离培养UC-MSCs的方法,培养的细胞为具有增殖能力强和更原始间充质干细胞,为建立间充质干细胞库和临床应用提供了理论依据。     

Notch信号介导共培养脐带间充质干细胞和造血干细胞的相互作用

目的:探讨脐带间充质干细胞(UC-MSC)体外与造血干细胞共培养后Notch信号分子的改变。方法:通过胶原酶消化方法分离UC-MSC,通过流式细胞仪检测以及成脂、成骨和成软骨诱导鉴定UC-MSC具备间充质干细胞的特性。进而,将UC-MSC与脐血CD34+造血干细胞(HSC)体外培养,实时PCR方法检测MSC及CD34+细胞表面Notch配体及受体表达以及表达是否存在变化;在共培养体系中加入Notch信号阻滞剂DAPT(γ-secretase抑制剂),比较Hes-1基因活化状态的改变。结果:体外实验显示:UC-MSC在形态学、细胞表面表型和诱导分化能力上均具备间充质干细胞的特性。UC-MSC及CD34+细胞表面存在Notch信号配体及受体的表达,共培养后Jagged 1、Notch1基因表达明显增加;共培养后CD34+细胞中的Hes-1基因表达明显增加而加入DAPT后Hes-1基因表达未检出明显改变。结论:UC-MSC支持造血中,Notch信号可能发挥重要作用。     

过表达IGF-1对脐带间充质干细胞氧化损伤的抑制作用

 目的: 分析过表达胰岛素样生长因子1(IGF-1)对脐带间充质干细胞(UC-MSCs)氧化损伤的抑制作用,开发UC-MSCs新的应用模式。方法: 通过酶消化法,从脐带组织中分离UC-MSCs,并采用流式细胞术对细胞的表面特异标志物进行鉴定,通过逆转录病毒感染体系,构建过表达IGF-1的UC-MSCs(UC-MSCs-IGF-1),并进一步通过实时荧光定量PCR和流式细胞术对UC-MSCs-IGF-1的IGF-1表达水平进行评价,同时分析UC-MSCs-IGF-1的表面标志物及成骨成脂分化能力;用不同浓度的H₂O₂(0 μmol/L、10 μmol/L、50 μmol/L和100 μmol/L)处理细胞,分析UC-MSCs-IGF-1在增殖活力维持、抗氧化损伤和抗凋亡方面的优势。结果: UC-MSCs表面标志物CD29、CD90和CD105的表达均呈阳性,而CD34的表达为阴性,符合正常间充质干细胞的表型特征;实时荧光定量PCR和流式细胞术结果表明,应用逆转录病毒感染体系构建的UC-MSCs-IGF-1在IGF-1表达方面远高于正常UC-MSCs,同时干细胞表型与UC-MSCs一致,且具备正常的成脂成骨分化能力;在H₂O₂的氧化损伤作用下,与UC-MSCs相比,UC-MSCs-IGF-1具有较强的增殖活力和抗氧化、抗凋亡功能,同时细胞中的SOD活性略高于UC-MSCs。结论: 过表达IGF-1对UC-MSCs的氧化损伤及氧化引起的凋亡均具有一定的防护作用,与正常UC-MSCs相比,UC-MSCs-IGF-1可能在临床应用方面具有一定的优势。     

脐带Wharton’s Jelly中间充质干细胞的生物学特性

背景：脐带Wharton’s Jelly中的间充质干细胞免疫原性低,增殖能力优于骨髓间充质干细胞,受到广泛关注。 目的：分离、培养人脐带Wharton’s Jelly中的间充质干细胞,并观测其生物学特性。 方法：利用组织块培养法分离培养脐带Wharton’s Jelly中的间充质干细胞,倒置显微镜下观察细胞形态,MTT法测定细胞倍增时间,流式细胞仪检测细胞免疫表型。 结果与结论：组织块培养第2天时,开始有细胞从组织块爬出;第7天细胞形态开始发生变化,部分细胞呈梭形。传至第3代的细胞形态均一,呈梭形或成纤维形。传至第7代、第10代的细胞形态无明显变化,仍呈梭形。MTT结果示细胞的倍增时间为三四天,传至10代后细胞倍增时间无明显差别。分离培养的脐带Wharton’s Jelly中间充质干细胞表达CD44、CD29、CD105,不表达CD34、CD45、CD14。     

人骨骼肌源性血管内皮细胞对造血干/祖细胞的体外支持

背景:人骨骼肌源性血管内皮细胞位于血管壁,共表达肌肉干细胞和血管内皮细胞的标记(CD56+CD34+CD144+CD45-).研究显示,人肌血管内皮细胞与间充质干细胞存在相似性,表达间充质干细胞表面标记物,具有多向分化潜能.目的:建立人肌血管内皮细胞作为滋养层与人脐血CD34+细胞体外培养体系,以培养前后CD34+细胞...     

人脐带间充质干细胞体外诱导分化为成纤维细胞

背景：脐带间充质干细胞具有多向分化能力,但其向成纤维细胞分化研究较少。 目的：验证人脐带间充质干细胞向成纤维细胞的分化能力。 方法：采用贴壁法分离脐带间充质干细胞,流式细胞仪分析其表面抗原。取第3代脐带间充质干细胞进行成脂成骨诱导分化,以碱性成纤维细胞生长因子诱导脐带间充质干细胞向成纤维细胞分化。 结果与结论：贴壁法能稳定从脐带中分离出干细胞,脐带间充质干细胞极低表达 CD31、CD45 、CD40、HLA-DR,强表达 CD29、CD90、CD44、CD105。脐带间充质干细胞成脂诱导后油红O染色显示胞浆中充满红色的油滴;成骨诱导后茜素红染色可在细胞密集区见红色的钙结节。碱性成纤维细胞生因子诱导后细胞表达Ⅰ型胶原明显高于对照组。提示贴壁法分离脐带间充质干细胞可靠、纯度高,碱性成纤维细胞生长因子可诱导脐带间充质干细胞向成纤维细胞分化。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：     

Umbilical cord blood mesenchymal stem cells

We studied umbilical cord blood mesenchymal stem cells and compared mesenchymal stem cells derived from umbilical cord blood, adipose tissue, and skin. Umbilical cord blood mesenchymal stem cells were characterized morphologically, cytofluorometrically, and by their differentiation potential. Umbilical cord blood mesenchymal stem cells did not differ from cells isolated from adipose tissue and skin by the main parameters (by morphology, expression of surface markers, and differentiation potential). A specific feature of umbilical cord blood mesenchymal stem cells is their low count per volume of the initial material and very low proliferative activity. __________ Translated from Kletochnye Tehnologii v Biologii i Medicine, No. 1, pp. 16–20, January, 2007     

Comparatine analysis of cytophenotypes of cells of mesenchymal lineage isolated from human tissues

The expression of cytoplasmic and surface proteins in cultured human skin fibroblasts, human umbilical cord cells obtained after normal delivery on gestation week 38–40, and mesenchymal bone marrow stem cells was compared by the methods of immunocytochemistry and flow cytofluorometry. Bone marrow mesenchymal stem cells expressed a great variety of marker proteins typical of stem and progenitor cells and did not express proteins typical of differentiated cells. Fibroblast-like umbilical cord cells expressed markers of both stem cells and differentiated cells. Fibroblasts of dermal origin were characterized by intensive expression of proteins typical of differentiated cells. __________ Translated from Kletochnye Tehnologii v Biologii i Medicine, No. 1, pp. 38–45, January, 2007     

不同培养基培养人脐血间充质干细胞的差异

背景：脐血间充质干细胞为干细胞领域的研究热点,但目前传代及扩增此类细胞尚无简单、有效、完美培养方法。 目的：采用不同的培养基分离培养融合状态的间充质干细胞,以筛选一种较好的体外培养人脐血间充质干细胞的方法。 方法：无菌条件下取正常足月剖宫产新生儿的脐血,随机分为5组：低糖DMEM(Dulbecco改良的Eagle培养基)组、高糖DMEM组、α-DMEM组、低糖DMEM+干细胞因子组、低糖DMEM +骨髓间充质干细胞培养上清组。用淋巴细胞分离液分离脐血的单个核细胞。将脐血单个核细胞接种于含体积分数为10%胎牛血清的上述培养基中,放置于37 ℃、体积分数为5%的CO2培养箱内培养,倒置显微镜观察细胞数量和形态的变化并用流式细胞技术分析细胞的表面抗原。 结果与结论：①各组间充质干细胞培养48 h后贴壁细胞数和细胞存活率的比较：低糖DMEM+干细胞因子组、低糖DMEM +骨髓间充质干细胞培养上清组的贴壁细胞数明显多于低糖DMEM组、高糖DMEM组、α-DMEM组(P < 0.05),细胞存活率亦明显高于低糖DMEM组、高糖DMEM组、α-DMEM组(P < 0.05)。②各组间充质干细胞在不同培养时间下生长状态的比较：培养第3,6,9,12,15,18,21天低糖DMEM+干细胞因子组、低糖DMEM+骨髓间充质干细胞培养上清组细胞增殖的速度均快于低糖DMEM组、高糖DMEM组、α-DMEM组(P < 0.05)。低糖DMEM+干细胞因子组与低糖DMEM +骨髓间充质干细胞培养上清组比较差异无显著性意义。结果可见人脐血间充质干细胞与骨髓间充质干细胞培养上清或干细胞因子共孵育,对脐血间充质干细胞体外分离培养及扩增有支持作用。     

Induction of acute graft vs. host disease in lymphopenic mice

Allogeneic hematopoietic stem cell transplantation (HSCT) is a potentially life-saving treatment for refractory/relapsing hematological malignancies, blood disorders or autoimmune diseases. However, approximately 40–50% of patients undergoing allogeneic HSCT will develop a multi-organ, inflammatory disorder called acute graft vs. host disease (aGVHD). Experimental and clinical studies suggest that intestinal injury due to toxic, pre-transplant conditioning protocols (e.g. lethal irradiation and/or chemotherapy) may play a major role in the development of aGVHD. However, recent studies from our laboratory suggest that this may not be the case. The objective of this study was to quantify and compare the onset and severity of aGVHD induced by the adoptive transfer of allogeneic T cells into untreated lymphopenic mice. Four million allogeneic or syngeneic CD4⁺CD62L⁺CD25⁻ T cells were transferred (i.p.) into NK cell-depleted RAG1^-/- mice or RAG2^-/-IL2rγ^-/- double knock-out (DKO) mice and assessed daily for signs of aGVHD. We found that adoptive transfer of allogeneic but not syngeneic T cells into NK cell-depleted RAG1^-/- or DKO mice induced many of the clinical and histological features of aGVHD including weight loss, inflammatory cytokine production and tissue inflammation. In addition, adoptive transfer of allogeneic T cells into each recipient induced severe anemia as well as dramatic reductions in bone marrow and spleen cellularity. Taken together, we conclude that allogeneic CD4⁺ T cells are both necessary and sufficient to induce aGVHD in lymphopenic recipients in the absence of toxic, pre-transplant conditioning.     

人脐带间充质干细胞与肝细胞共培养可分化为肝样细胞

背景：文献报道,从骨髓与脐带中分离获得的间充质干细胞可在体外连续传代培养,仍保持干细胞的特性,并在多种细胞因子的“鸡尾酒式”诱导下分化为肝细胞样细胞。 目的：进一步验证人脐带间充质干细胞在体外正常人肝细胞共培养体系下是否可分化为肝细胞并探讨其分化方法。 方法：采用贴壁法,从脐带中分离培养间充质干细胞,流式细胞仪检测脐带间充质干细胞表面标志。人肝细胞LO2细胞与人脐带间充质干细胞建立共培养体系,不添加外源诱导因子,分别于第7,14,21天,通过RT-PCR 法检测肝细胞特异标志物甲胎蛋白、白蛋白、人细胞角蛋白19 mRNA的表达,糖原染色进行功能鉴定。 结果与结论：从人脐带中可分离得到贴壁生长的间充质干细胞,其中CD29⁺细胞比例为96.02%,CD105⁺细胞比例为96.6%,CD34^-细胞比例为99.65%,CD105⁺CD29⁺双阳性细胞比例为94.84%。与LO2细胞共培养后第7天仅有甲胎蛋白阳性表达;第14天表达白蛋白、人细胞角蛋白19,第21天时,LO2与人脐带间充质干细胞共培养组未出现甲胎蛋白表达;人细胞角蛋白19和白蛋白的表达比第14天略有增强。共培养21 d后,糖原染色呈阳性。结果证实,无需额外添加外源诱导因子,脐带间充质干细胞可在人正常肝细胞共培养的微环境中,向正常肝细胞分化。     

人脐带间充质干细胞的分离鉴定及多向诱导分化

背景：脐带间充质干细胞取材方便、无创,不受伦理学限制,比一般干细胞原始,免疫原性小,其应用前景广阔,是一种理想的种子细胞。 目的：分离鉴定脐带间充质干细胞,并诱导其向成骨细胞和成脂细胞分化。 方法：组织块贴壁法分离纯化脐带间充质干细胞,取对数生长期的第3代细胞,观察细胞形态、生长方式;流式细胞仪检测干细胞表型CD90、CD105、CD34和CD45的表达情况,并在体外检测能否将其诱导分化为成脂细胞及成骨细胞。 结果与结论：用组织块法成功分离培养出脐带间充质干细胞,流式细胞学鉴定显示细胞强表达CD90和CD105,不表达CD34和CD45;能在体外将其成功诱导为脂肪细胞和成骨细胞。结果显示组织块贴壁法能够从人脐带中分离出间充质干细胞,该细胞可向成脂细胞及成骨细胞分化。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：     

脐血来源间充质干细胞可抑制异体T细胞的反应

目的研究脐血间充质干细胞(HUCB-MSCs)对异体T细胞的抑制作用。方法体外培养HUCB-MSCs,流式细胞术测表面标记;取正常人外周血,免疫磁珠分离CD3+T细胞,将分离的CD3+T与HUCB-MSCs 1∶1混合培养5 d,PHA刺激或不刺激,采用3H-TdR掺入法观察T细胞增殖,ELISA方法检测细胞因子,流式细胞术观察细胞凋亡。结果 HUCB-MSCs呈纺锤样的细胞形态,不表达CD14、CD34、CD45、HLA-DR,而表达CD29、CD44、HLA-ABC。HUCB-MSCs抑制PHA引起的T细胞增殖(5 230±550 vs 10 500±800 counts/min,P<0.001);HUCB-MSCs还能抑制异体T细胞分泌IFN-γ(510±60 vs 1 580±100 pg/mL,P<0.001)和TNF-α(590±20 vs 1 180±30 pg/mL,P<0.001),上调IL-4(16.3±8.2 vs 4.1±1.8 pg/mL,P<0.001)和IL-10(105±5 vs 17±2 pg/mL,P<0.001)分泌;HUCB-MSCs不诱导T细胞的凋亡。结论 HUCB-...     

冻存脐带间充质干细胞向成骨细胞的分化

背景：在骨组织工程中,脐带间充质干细胞是的一种新兴的种子细胞。目前认为低温冻存是长期保存细胞的有效方法。 目的：探究冻存的脐带间充质干细胞能否被诱导分化成成骨细胞。 方法：采用组织块贴壁法从脐带的华尔通氏胶组织中分离出间充质干细胞。然后,用倒置显微镜观察原代细胞的细胞形态。脐带间充质干细胞的免疫表型和细胞周期用流式细胞仪检测。在冻存6个月后,复苏第2代脐带间充质干细胞进行冻存复苏,并传代培养至12代。对第12代的脐带间充质干细胞进行成骨诱导,它的成骨能力分别通过碱性磷酸酶活性检测,骨钙素和骨涎蛋白的免疫荧光检测以及茜素红染色检测来 确定。 结果与结论：原代脐带间充质干细胞呈现典型的成纤维细胞样形态。流式细胞仪显示培养的细胞高表达间充质干细胞的表面标志CD73、CD105和CD90,但是不表达造血细胞的表面标志CD34和CD45。复苏后细胞的存活率是90%。细胞周期显示P8的细胞有75%处于G₀/G₁期,25%处于S+G₂M期。经成骨诱导液处理的第12代细胞显示出比对照组更高的碱性磷酸酶活性(P < 0.01)。此外,在成骨诱导液中诱导的细胞对骨钙素和骨涎蛋白的染色呈阳性,并形成矿化了结节。冻存后的脐带间充质干细胞仍保持了它们的生物学特性,并且在成骨诱导液中能被诱导分化成成骨细胞。