脐带间充质干细胞修复皮肤创伤的研究进展

脐带间充质干细胞(UC-MSCs)是一种免疫原性低、易于获取和扩增的成体干细胞。UC-MSCs可以调节伤口炎性反应水平,促进血管形成、细胞增殖,抑制瘢痕形成,诱导皮肤附件形成。UC-MSCs联合其他药物作用于伤口可以促进UC-MSCs的黏附、存活、迁移和增殖等。     

人脐带间充质干细胞的研究进展

脐带间充质干细胞是存在于脐带沃顿胶和血管周围组织中的一种干细胞,具有多向分化和自我更新的潜能。脐带间充质干细胞可分化为骨细胞、软骨细胞、脂肪细胞、肌细胞和神经细胞,并且具有免疫调节性;脐带作为医疗废弃物来源丰富,对供者无不利影响,无伦理问题的限制,这为细胞治疗和组织工程提供了新的种子细胞。     

脐带间充质干细胞与系统性红斑狼疮

系统性红斑狼疮（SLE）是一种以免疫细胞活化和自身抗体产生为特征的自身免疫病。间充质干细胞（MSC）是属于中胚层的一类多能干细胞,不仅具有多向分化潜能,还有多种免疫调节作用。许多研究表明MSC对可以通过调节免疫细胞（上调Foxp3’T细胞水平、下调Th17细胞水平、抑制浆细胞的成熟等）的表达以及细胞因子（IL-10、TGF—β、TNF—α等）的分泌干预抑制SLE,并且免疫原性较低。尤其脐带MSC,因为其独特优势,目前已经成为干细胞干预治疗SLE的研究热点。     

人脐带间充质干细胞体外分泌的伤口愈合相关因子

背景：目前对人脐带间充质干细胞培养上清液中伤口愈合相关因子定量检测研究的报道较少。 目的：检测人脐带间充质干细胞培养上清中伤口愈合相关因子白细胞介素6,白细胞介素8,转化生长因子1,单核细胞趋化蛋白1,血管内皮生长因子,GM-CSF和TIMP-1的分泌水平。 方法：以胎牛血清替代物CCS1107为培养基添加物培养人脐带间充质干细胞。将培养获得的P2代细胞以1.1×104/cm2的接种密度接种于直径100 mm无菌培养皿中,培养96 h后,收集上清液并对其中所含伤口愈合相关因子进行ELISA定量检测。 结果与结论：10份不同个体间的因子分泌量差异无显著性意义,男女间表达量差异无显著性意义。高表达白细胞介素6, 白细胞介素8,单核细胞趋化蛋白1和组织基质金属蛋白酶抑制剂1,其中单核细胞趋化蛋白1和组织基质金属蛋白酶抑制剂1的表达量最高。结果证实,人脐带间充质干细胞培养上清液中含有大量伤口愈合相关因子。     

脐带间充质干细胞与系统性红斑狼疮

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the activation of immune cells and production of autoantibodies by plasma cells and release of cytokines. Mesenchymal stem cells (MSC) are widely studied as an alternative cell source for their ability to differentiate into multiple mesenchymal lineages. An important function for MSC for autoimmune diseases is their immunomodulatory effect on various activated lymphoid cells, such as T cells B cells, NK cells, and dendritic cells; and on cytokines , such as IL-10, TGF-β, TNF-α, et al. Umbilical cord MSCs (UC-MSCs) had a higher proliferation capacity and lower immunogenicity, indicating that it may be a novel alternative source of human MSC for clinical application. In this review, we introduce the immunomodulatory effect and clinical application of UC-MSC in SLE based on recent findings in human and animal models.     

胶原-壳聚糖支架材料与间充质干细胞的组织相容性

背景：近年来一些研究发现胶原蛋白-壳聚糖复合支架材料可作为神经组织工程的支架材料,但相关细胞相容性研究较少。 目的：观察兔骨髓间充质干细胞在胶原蛋白-壳聚糖复合支架材料表面生长及分化情况。 方法：分离培养兔骨髓间充质干细胞,无血清培养液培养,流式细胞仪检查细胞表型;然后,将其接种到凝胶支架材料表面(实验组)及多聚赖氨酸包被的盖玻片表面(对照组),神经诱导培养基内培养,倒置相差显微镜观察干细胞的生长及分化情况。 结果与结论：细胞表型为CD29⁺、CD44⁺、CD166⁺。倒置相差显微镜观察：实验组中,接种的骨髓间充质干细胞生长良好,7 d后可见有突起神经细胞,细胞生长情况与对照组未见有明显差别。证实胶原蛋白-壳聚糖复合支架材料对骨髓间充质干细胞有良好细胞相容性。     

脐带间充质干细胞治疗脊髓损伤临床分析

目的观察脐带间充质干细胞（UC-MSCs）鞘内注射治疗脊髓损伤（SCI）的临床效果及安全性。方法对2008年1月至2010年10月收治的22例SCI患者,给予UC—MSCs鞘内注射治疗,细胞数1×10^6个／（kg·次）,1次／周,4次为1个疗程,其中4例接受2个疗程,1例接受3个疗程,余均接受1个疗程。采用美国脊髓损伤协会制定的脊髓损伤神经功能评分标准（ASIA标准）对患者治疗前后神经功能进行评定,采用国际神经修复学会脊髓损伤功能评价量表（IANR—SCIRFS）对患者治疗前后日常生活活动能力进行评定。结果22例患者中13例有效,9例无效。不完全性SCI患者有效率达81．25％,完全性SCI的6例患者均无效。有5例有效的患者接受了2～3个疗程治疗,疗效均有进一步的提高。有效患者多表现为运动和／或感觉功能改善,大小便控制能力增强。22例患者治疗后1个月与治疗前比较,痛觉、触觉、运动、日常生活活动能力评分均有明显升高（P〈0．01）。治疗后常见的不良反应有头痛（1例）、腰痛（1例）,均在1～3d内消失。随访3个月至3年,无治疗相关不良事件发生。结论UC—MSCs鞘内注射治疗是安全的,可以改善大部分不完全性SCI患者的神经功能,提高这些患者的生活质量。     

人脐带间充质干细胞的生物学性质研究进展

人脐带中富含间充质干细胞(MSCs)。这些细胞能表达多种间充质干细胞标志物及多种干细胞相关基因,能分化为3个胚层衍生的多种成熟细胞,合成多种营养因子和细胞因子,支持造血干细胞等细胞的增殖和功能,并具有低免疫原性。     

人脐带间充质干细胞肌肉注射健康大鼠心肌组织微血管及胶原纤维的表达

BACKGROUND:To date, it is still unclear whether the intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells (UC-MSC) can cause cardiac ectopic pathological angiogenesis as well as increase collagen synthesis to promote myocardial fibrosis. OBJECTIVE:To explore the effects of intramuscular injection of human UC-MSCs on myocardial micrangium and collagen expression in normal Wistar rats. METHODS:After 2 weeks of feeding, 60 male SPF Wistar rats were randomly assigned to receive intramuscular injection of PBS (normal group), DMEM (culture medium group), human UC-MSCs supernatant (supernatant group), 0.25×10⁵, 1.0×10⁵, 4.0×10⁵ human UC-MSCs (low-, moderate- and high-dose groups), respectively (n=10 per group). All the rats were subjected to second injection (same dose) at 4 weeks after first intramuscular injection. Then, the rats were killed under anesthesia at 4 weeks after second injection, to take heart tissues from the left ventricle for pathological observation, immunohistochemical examination and Masson staining. RESULTS AND CONCLUSION: No alteration of the response, activity, victualage, faeces, weight growth, and fur was found, and there was no death in rats during the experiment. All the rats had no symptoms of molt, inflammation, skin ulcer, scleroma. Strong positive expression of CD34 for the micrangium in the myocardial tissue was observed, and positive expression of the collagen in the myocardial tissue observed by Masson staining. There were no significant differences in the microvessel density and collagen expression in the myocardium among the groups (F=0.110 and 0.585, P > 0.05). To conclude, hUC-MSCs or its supernatant via intramuscular injection has no effect on the micrangium and collagen expression in normal rats.     

脐带间充质干细胞培养中的染色标记及示踪技术

背景：脐带间充质干细胞的培养是进行脐带间充质干细胞研究时极其重要的部分,细胞培养技术的优化对推动间充质干细胞的临床应用特别是细胞治疗至关重要。同时,脐带间充质干细胞标记及示踪技术是干细胞移植治疗的研究热点之一。目的：对国内外脐带间充质干细胞染色标记技术的研究与进展作一综述。方法：以“干细胞,间充质干细胞,脐带间充质干细胞,细胞培养,染色标记”为中文捡索词,以“Umbilical cord-derived mesenchymalstem cells,Cell culture,Labeling methods”为英文检索词,检索维普和中国知网(CNKI)期刊全文数据库、Medline,highwire和外文生物医学期刊全文数据库(Foreign Journals Integration System)2001年1月至2013年10月有关脐带间充质干细胞的培养及标记染色文献。最终纳入35篇文献进行综述。结果与结论：脐带间充质干细胞尚未得到广泛应用,最关键的原因是脐带间充质干细胞的分离培养及染色技术不是很成熟,这些方面都值得在研究中进一步优化。虽然近几年脐带间充质干细胞标记及示踪技术有了较快的进展,但仍存在许多问题有待进一步解决。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：     

银屑病患者与正常人皮肤间充质干细胞的生物学特性

BACKGROUND:Skin-derived mesenchymal stem cells may reflect the onset of psoriasis. OBJECTIVE:To analyze the biological characteristics of skin-derived mesenchymal stem cells in psoriasis patients. METHODS:Skin-derived mesenchymal stem cells from 30 patients with psoriasis and 20 healthy controls were isolated and cultured by trypsin. Flow cytometry was used to detect the cellular immune phenotypes CD34, CD44, CD29, CD45, CD90, CD105, CD73 and HLA-DR. The mesenchymal stem cells were induced by the corresponding cartilage, osteogenic and osteogenic inducing agents, to identify the multi-directional differentiation ability. The cell proliferation curve was plotted at passage 3, and the levels of transforming growth factor-β1 and epidermal growth factor in culture supernatant were detected by ELISA assay. RESULTS AND CONCLUSION:Under an inverted phase contrast microscope, primary skin-derived mesenchymal stem cells isolated from patients with psoriasis and normal controls both exhibited heterogeneity. In the two groups, CD29, CD90, CD44, CD73 and CD105 were highly expressed, and CD45, CD34, and HLA-DR were lowly expressed. Under certain conditions, skin-derived mesenchymal stem cells were induced to differentiate into adipocytes, osteoblasts or chondrocytes. Proliferation of skin-derived mesenchymal stem cells in the psoriasis group was significantly faster than that in control group, but the final number of cells in the two groups tended to be consistent. The levels of transforming growth factor-β1 and epidermal growth factor in the psoriatic skin had no correlation with the severity of the disease (P > 0.05). Compared with the control group, the epidermal growth factor level in the cell supernatant was significantly higher in the psoriasis group (P < 0.01), while the level of transforming growth factor-β1 was significantly lower (P < 0.01). These results showed that there is heterogeneity in the morphology of skin-derived mesenchymal stem cells from psoriasis patients, and the biological activity of mesenchymal stem cells is abnormal.     

毛囊干细胞在皮肤伤口修复中的促进作用

背景：毛囊干细胞来源于皮肤、毛发,数量极其可观,取材后无严重的并发症和免疫原性,可供自体移植,是最容易获取的干细胞来源之一。目的：综述毛囊干细胞促进皮肤伤口修复的作用因素,以期为该领域研究提供有效的实验依据。方法：应用计算机以“毛囊干细胞、皮肤修复、再生医学、组织工程”或“hair follicle stem cell,skin repairing,regenerative medicine,tissue engineering”为检索词检索CNKI 和 PubMed 数据库1999年1月至2014年12月发表的关于毛囊干细胞促进皮肤伤口修复的文章,最终选择45篇文章进行综述。结果与结论：毛囊干细胞作为一种容易获得、数量可观、使用较为安全以及具有分化潜能的成体干细胞,可以通过皮肤早期血管化、表皮及附属器官的再生、信号通路作用及转录因子调节等方面的影响,促进皮肤伤口的修复,能为再生医学及组织工程研究提供良好的种子细胞。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：     

循环纤维细胞与慢性创面愈合

背景：循环纤维细胞是近些年来在外周血液发现的具有成纤维细胞特性的一种白细胞亚群,由于具有合成多种细胞外基质蛋白、细胞因子以及递呈抗原、收缩创面、促进新生血管形成的能力,因此被认为可以促进创伤的修复。但其促进慢性创面修复的潜在作用研究尚少。 目的：通过文献检索,对循环纤维细胞的生物学特性及其在慢性创面修复中的潜在作用进行文献综述。 方法：分别以“循环纤维细胞、慢性创面、糖尿病足、创面愈合、细胞治疗”和“circulating fibrocytes、An-healing wounds、diabetic foot ulcer、wound healing、cell therapy”为关键词进行检索,CNKI数据库的检索时限为2000至2014年,PubMed数据库的检索时限为1994至2015年,西文生物医学期刊文献数据的检索时限为2000至2015年,检索内容为循环纤维细胞、慢性创面的难愈机制以及细胞治疗在慢性创面愈合中的应用。保留符合纳入标准的54篇文献进行总结分析。 结果与结论：循环纤维细胞因其安全、有效并能较好的发挥促进创面愈合的作用,细胞治疗已开始应用于创面修复。循环纤维细胞是在外周血发现的具有成纤维细胞特性的一个新型白细胞亚群,具有合成多种细胞外基质蛋白、细胞因子以及递呈抗原、收缩创面、促进新生血管形成的能力并在伤后早期进入损伤部位,在创伤修复过程中发挥着积极作用。动物研究证实,应用循环纤维细胞可改善慢性创面尤其是糖尿病慢性创面的修复。   中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

不同培养方法对人脐带间充质干细胞的影响

BACKGROUND:Umbilical cord stem cells mainly derive from full-term infants, and common culture methods include tissue-attached method and trypsin-digestion mehod. However, effects of different culture methods on the separation of umbilical cord mesenchymal stem cells remain many disputes. OBJECTIVE:To observe the effects of different culture methods on umbilical cord mesenchymal stem cells. METHODS:Umbilical cords of 30 healthy full-term and caesarean delivery infants were selected, and cultured using tissue-attached method or trypsin-digestion method to isloate and culture human umbilical cord mesenchymal stem cells. Meanwhile, cell growth was measured by flow cytometry. RESULTS AND CONCLUSION:The fusiform-shaped cells began to separate from the umbilical cord tissue that was primary cultured using tissue-attached method, and 10 days later, the cell fusion reached 80%; after the umbilical cord was cultured using collagenase-trypsin digestion for 5 days, a small amount of adherent cells with different shapes appeared, and the fiber-like cells reached 80% of confluence until 2-week culture. There was no significant difference in the growth of umbilical cord mesenchymal stem cells cultured by different culture methods (P > 0.05). Moreover, cells cultured by two methods were all positive for CD13, CD29, CD44 and CD105. These results demonstrate that human umbilical cord mesenchymal stem cells exhibit a high success rate in primary culture using tissue-attached method, which is superior to the trypsin-digestion method.     

损伤皮肤修复和伤口愈合过程中的干细胞：问题与前景

BACKGROUND:Present treatments for chronic skin wounds have certain limitations, and adult stem cells play a potential part in cutaneous repair and regeneration. OBJECTIVE:To review effects of stem cells in skin regeneration and wound healing. METHODS:The first author retrieved CNKI and Medline databases by computer for relevant articles published from 2000 to 2010. The keywords were “epidermal stem cells, hair follicle stem cells, stem cells, transplantation, dermal stem cells” in Chinese and in English, respectively. Then totally 489 papers were obtained after initial survey, and according to the inclusion criteria, 30 articles were selected for review. RESULTS AND CONCLUSION:Epidermal stem cells and other adult stem cells have been applied to treat wounds and other skin diseases. Epidermal stem cells are the crucial cell source of skin development, repair and remodeling. Epidermal stem cells are always in a resting state in vivo. Unless, skin injure or culture in vitro, cell division and proliferation will be significantly fastened. The stability of the epidermis mainly depends on the asymmetric division of a subpopulation, in which two daughter cells are produced, including one with characteristics of stem cells, and the other differentiated into transient amplifying cells that will be differentiated into post mitotic cells after a series of cell divisions (3-5 times). Afterwards, those post mitotic cells are developed into terminal differentiation cells on the basal layer, finally detach from the epidermis as dander. In addition, it is unclear whether epidermal factors are related to apoptosis, migration and differentiation in the process of wound repair and even under physiological conditions. Therefore, application of stem cells in wound healing requires a further discussion.     

皮肤成纤维细胞在创面愈合中的研究进展

皮肤成纤维细胞是参与创面愈合的主要修复细胞,近年来其异质性及其与周围细胞间的通讯正逐渐引起重视.真皮成纤维细胞亚群主要包括乳头状成纤维细胞和网状成纤维细胞,对创面愈合发挥不同的作用.成纤维细胞通过自分泌和旁分泌信号分子与周围细胞之间相互作用构成创面微环境,影响创面愈合.慢性创面中的成纤维细胞表现出多种功能障碍.本文就成...     

富血小板纤维蛋白与胶原膜修复牙龈缺损创面的能力

BACKGROUND: Previous studies have found that platelet-rich fibrin has a good ability to induce gingival soft tissue repair and regeneration. OBJECTIVE: To observe the effects of platelet-rich fibrin versus collagen membrane on gingival soft tissue healing, and to evaluate the ability of platelet-rich fibrin to repair gingival defects. METHODS: Twenty-two patients (2 premolar teeth and 20 molars) scheduled for premolar or molar removal or ridge preservation due to various reasons were selected and randomized into two groups. Bio-Oss was implanted into the extraction socket followed by covering with platelet-rich fibrin or collagen membrane. Healing time and healing rate of gingival defects were detected to evaluate the ability of platelet-rich fibrin to promote gingival tissue healing at 1-2 weeks after Bio-Oss implantation. RESULTS AND CONCLUSION: The healing time was (12.17±2.25) days in the platelet-rich fibrin group and (17.30±2.58) days in the collagen group. The healing rate of the platelet-rich fibrin group was notably higher than that in the collagen membrane group at 1 and 2 weeks after Bio-Oss implantation. These findings indicate that platelet-rich fibrin is better than collagen membrane to improve the healing of gingival soft tissues with a shorter healing time. 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

骨髓间充质干细胞对创伤性骨折愈合的促进作用

BACKGROUND:In recent years, the development of stem cell culture and isolation technologies provides new therapeutic choices for fracture healing. OBJECTIVE:To investigate the effect of exogenous bone marrow mesenchymal stem cells on bone fracture healing in traumatic fracture rats and on the migration ability of endogenous bone marrow mesenchymal stem cells. METHODS:Femoral fracture models were made in 48 Wistar rats and then randomized into experimental group and control group (n=24/group). Bone marrow mesenchymal stem cells from another healthy rats were isolated using adherent method and then injected into the rats via the tail vein in the experimental group. Rats in the control group were given the same volume of normal saline. At 2, 3, 4, 8, 12 weeks after injection, we extracted bone marrow mesenchymal stem cells from the femur of rats in the two groups. RT-qPCR was used to detect expression levels of type I collagen and CD44. Transwell method was used to detect cell migration ability. Immunohistochemitry method was employed to detect expression of nerve growth factors in the callus. RESULTS AND CONCLUSION:mRNA levels of type I collagen and CD44 in rat bone marrow mesenchymal stem cells were significantly higher in the experimental group than the control group at 2, 3 and 4 weeks after injection (P < 0.05). Compared with the control group, the higher migration ability of bone marrow mesenchymal stem cells was found in the experimental group at 2 and 3 weeks after injection (P < 0.05) as well as the higher expression of nerve growth factor in the callus in the experimental group at 3, 4, 8, 12 weeks after injection. All these findings suggest that exogenous bone marrow mesenchymal stem cells can improve the migration ability of endogenous bone marrow mesenchymal stem cells and the expression of nerve growth factor in the callus in rats with femoral fracture, thereby promoting fracture healing in rats.     

脐血间充质干细胞移植在卵巢癌化疗损伤中的作用

背景:研究证实,脐血间充质干细胞对多种损伤细胞有修复功能。目的:探讨脐血间充质干细胞移植在卵巢癌化疗性损伤中的作用。方法:健康级成年雌性Wistar大鼠60只,随机分为正常对照组,损伤组及治疗组,每组20只。对照组不做任何处理,损伤组及治疗组建立大鼠卵巢癌化疗性损伤模型。模型成功后对照组尾静脉注射生理盐水,损伤组注射紫杉醇,治疗组在损伤组基础上给予脐血间充质干细胞移植。移植后2周,应用RT-PCR检测卵巢癌组织XAF1、Survivin mR NA的表达,Western blot检测卵巢癌组织中XAF1、Survivin蛋白的表达,TUNEL法检测卵巢癌细胞凋亡情况。结果与结论:(1)与损伤组比较,治疗组肿瘤组织XAF1 mR NA和蛋白表达显著上调,凋亡抑制蛋白Survivin mR NA和蛋白表达显著下调(P<0.05);(2)损伤组卵巢组织结构较对照组明显破坏,有大面积的出血坏死区域,治疗组损伤程度较损伤组明显减轻;(3)与损伤组比较,治疗组卵巢癌细胞凋亡率明显升高(P<0.05);(4)结果表明,脐血间充质干细胞移植可以促进卵巢癌化疗性损伤大鼠卵巢组织的修复,XAF1、Survivin在肿瘤新生血管形成及卵巢癌细胞凋亡过程中发挥了作用。     

盐霉素联合脐血间充质干细胞干预对乳腺癌MCF-7细胞的影响及作用机制

BACKGROUND:Mesenchymal stem cells-released factors can form a microenvironment inhibiting malignant tumor cell proliferation, and moreover, salinomycin also inhibits the growth of tumor cells. OBJECTIVE:To explore the effect of salinomycin with umbilical cord blood mesenchymal stem cells in breast cancer MCF-7 cells and its action mechanism, in order to seek a new target and therapeutic strategy to treat breast cancer. METHODS:Logarithmically growing MCF-7 cells were randomly divided into control, umbilical cord blood mesenchymal stem cell group and combination group. Cells in the control group were given no treatment, while those in the other two groups were given umbilical cord blood mesenchymal stem cell suspension or umbilical cord blood mesenchymal stem cell suspension combined with salinomycin, respectively. RESULTS AND CONCLUSION:Forty-eight hours after the intervention, the proliferation and invasion of MCF-7 cells and expression of POSTN protein in cells were significantly reduced in the combination group compared with the other two groups. These findings indicate that the combination of umbilical cord blood mesenchymal stem cell suspension and salinomycin effectively reduces the proliferation and migration of MCF-7 cells, and this combined use provides a new insight into the treatment of breast cancer.