脂肪间充质干细胞在系统性硬化症中的研究现状与展望

间充质干细胞(mesenchymal stromal/stem cells,MSCs)是一种非造血多能干细胞。最近,研究发现MSCs具有免疫调节、血管生成和抗纤维化的能力,从而使以MSCs为基础的干细胞应用逐渐从再生医学领域转向对自身免疫性疾病的治疗研究。脂肪间充质干细胞(adipose mesenchymal stem cells,ASCs)是来源于脂肪组织的MSCs。该文回顾当前MSCs在系统性硬化症(systemic sclerosis,SSc)中的研究现状,并展望临床应用ASCs治疗SSc的可行性。     

间充质干细胞治疗特应性皮炎的研究进展

特应性皮炎(AD)是一种复杂的慢性及复发性炎症性疾病,确切发病机制仍然不清楚,但其发展是遗传、免疫细胞及神经内分泌等多种系统相互作用的结果。许多细胞被证明参与AD的发病机制,其中干细胞,特别是间充质干细胞(MSCs)具有多项分化和免疫调节作用,其中包括调控T细胞、B细胞以及其他炎性细胞和炎症因子等,对AD有潜在的治疗作用。该文对MSCs在AD治疗中的作用机制作一综述。     

红斑、丘疹及鳞屑性皮肤病

20131750银屑病患者皮损间充质干细胞细胞因子的分泌水平/闫鑫(山西医科大学),刘瑞风,侯瑞霞…//中国皮肤性病学杂志.-2013,27(6).-553~556分离患者皮损与健康人皮肤皮损间充质干细胞(MSCs),流式细胞术及多向分化法进行细胞鉴定;ELISA法检测第3代皮肤MSCs培养上清液白细胞介素-8和粒细胞-巨噬细胞集落刺激因子的分泌水平。结果:两组MSCs的细胞形态、表面标志和多向分化能力相似,细胞标记CD29,D90及CD105均呈高表达,而     

间充质干细胞移植治疗系统性红斑狼疮的研究进展

间充质干细胞是来源于发育早期中胚层的成体干细胞,具有高度增殖、自我更新能力、多向分化潜能、损伤组织修复功能和免疫抑制能力。该文综述间充质干细胞,主要是脐带间充质干细胞和骨髓间充质干细胞用于治疗系统性红斑狼疮的研究进展,并提示间充质干细胞移植在未来的细胞治疗中是非常有前景的一种方法。     

人表皮干细胞的体外培养与MSCs对其诱导的初步研究

目的：探讨人表皮干细胞体外分离培养的理想方法，初步研究骨髓间充质干细胞（MSCs）上清对其生长的影响。方法：采用中性蛋白酶-胰蛋白酶两步法收获表皮细胞，Ⅳ型胶原快速粘附分选表皮干细胞，用无血清培养基进行体外培养，采用SABC—FITC进行培养细胞鉴定。结果：胎儿皮肤和人包皮皮肤均可成功分离到表皮干细胞集落，且增殖能力强，细胞表达Bl整合素和角蛋白CK19。MSCs上清形成的条件培养基的诱导下，收集的表皮细胞形成花环样克隆，并形成成纤维样细胞。结论：用酶消化法、胶原快速粘附和无血清培养法是较理想的体外分离培养表皮干细胞的方法，MSCs可以诱导表皮细胞的定向分化。     

HaCaT细胞诱导兔骨髓间充质干细胞表达角蛋白的初步研究

目的:探讨在体外培养的条件下HaCaT细胞诱导兔骨髓间充质干细胞(MSCs)表达角蛋白的可能性。方法:体外分离培养MSCs,流式细胞术检测CD34、CD45和CD44蛋白的表达以鉴定MSCs细胞。传代后与HaCaT共培养。分别在共培养3、7天后,密度梯度离心法分离MSCs和HaCaT,免疫细胞化学方法检测角蛋白Pan-cytokeratin(Pan-CK)、CK19的表达。结果:流式细胞术检测体外培养的MSCs CD34、CD45阴性,CD44阳性,表明MSCs培养成功。MSCs与HaCaT共培养3天、7天后,免疫细胞化学检测结果显示Pan-CK和CK19均为阳性。共培养7天的MSCs Pan-CK阳性率明显高于共培养3天的MSCs[分别为(29.1±8.3)%、(13.5±3.7)%,t=13.35,P0.05],共培养7天的MSCs CK19阳性率与共培养3天的MSCs相比无统计学差异[分别为(3.3±0.8)%、(3.1±0.5)%,t=1.37,P0.05]。结论:与HaCaT细胞共培养可诱导兔骨髓间充质干细胞向表皮细胞及表皮干细胞分化。     

HaCaT细胞诱导兔骨髓间充质干细胞表达角蛋白的初步研究

目的:探讨在体外培养的条件下HaCaT细胞诱导兔骨髓间充质干细胞(MSCs)表达角蛋白的可能性。方法:体外分离培养MSCs,流式细胞术检测CD34、CD45和CD44蛋白的表达以鉴定MSCs细胞。传代后与HaCaT共培养。分别在共培养3、7天后,密度梯度离心法分离MSCs和HaCaT,免疫细胞化学方法检测角蛋白Pan-cytokeratin(Pan-CK)、CK19的表达。结果:流式细胞术检测体外培养的MSCs CD34、CD45阴性,CD44阳性,表明MSCs培养成功。MSCs与HaCaT共培养3天、7天后,免疫细胞化学检测结果显示Pan-CK和CK19均为阳性。共培养7天的MSCs Pan-CK阳性率明显高于共培养3天的MSCs[分别为(29.1±8.3)%、(13.5±3.7)%,t=13.35,P<0.05],共培养7天的MSCs CK19阳性率与共培养3天的MSCs相比无统计学差异[分别为(3.3±0.8)%、(3.1±0.5)%,t=1.37,P>0.05]。结论:与HaCaT细胞共培养可诱导兔骨髓间充质干细胞向表皮细胞及表皮干细胞分化。     

皮肤非结核分枝杆菌感染2例报告

背景：非结核分枝杆菌的培养比较困难，国内关于确诊非结核分枝杆菌感染的报告很少，目的：探讨非结核分枝杆菌感染的实验室诊断方法及非结核分枝杆菌感染的治疗措施。方法：应用聚合酶链反应和ELISA技术进行实验室诊断，并以联合疗法分别治疗之。结果：2例非结核分枝杆菌感染患者经联合疗法治疗有显著疗效。结论：所用的聚合酶链反应和ELISA有助于诊断非结核分枝杆菌感染，氟嗪酸、利福平、氨苯砜联合疗法治疗有显著疗效。     

皮肤间充质干细胞在促进皮肤愈合中的作用

间充质干细胞(mesenchymal stem cells,MSCs)是当前干细胞研究的热点之一。目前,皮肤愈合正逐渐受到重视。现有的研究认为骨髓间充质干细胞(BM-MSCs)能从多个方面促进皮肤愈合,如促进表皮生长、促进真皮成纤维细胞的增生等。皮肤间充质干细胞(SMSCs)和BM-MSCs均为MSCs,具有很多的相似性,且SMSCs较BM-MSCs更容易得到。所以可从目前对BM-MSCs的研究预测到SMSCs在皮肤创伤愈合中的研究前景,且将来很可能会替代BM-MSCs。     

干细胞移植治疗与重要脏器功能的修复与重建

干细胞（stem cells, SCs）是具有自我更新、高度增殖和多向分化潜能的细胞，通过分裂维持自身细胞群的大小，同时又可分化成为各种不同的组织细胞，从而构成机体各种复杂的组织器官[1]。根据干细胞所处的发育阶段分为：胚胎干细胞（embryonic stem cells，ESCs）；成体干细胞（somatic stem cells，SSCs）。人体几乎所有组织都存在经历相当程度的分化SSCs，但在特定条件下，一种组织的SSCs可以“横向分化”成其他组织的功能细胞，如间充质干细胞（mesenchymal stem cells, MSCs）可以分化成神经、肌肉、软骨和骨等多种细胞[2]。MSCs属于中胚层的一类多能干细胞，主要存在于结缔组织和器官间质中，以骨髓组织中含量最为丰富，且主要来源于骨髓，又称为骨髓间充质干细胞（bone marrow mesenchymal stem cells, BMSCs）。Friedenstein等[3]于1976年首先分离BMSCs，研究证实BMSCs具有强大的增殖能力和多向分化潜能、免疫调节及免疫重建功能，并且来源方便，易于分离、培养、扩增和纯化，经多次传代扩增后仍具有干细胞特性，不存在免疫排斥。因此，BMSCs可能成为细胞移植和基因治疗的理想材料，目前国内外正在探索用于神经系统疾病及多领域多学科难治性疾病的治疗，并已取得了重大突破性进展[4-6]。我国MSCs的研究发展极为迅速，某些核心技术已占国际领先地位，2001年国家批准建立“国家干细胞工程产品产业化基地”，2002年4月在天津建立“国家干细胞工程技术研究中心”。     

Cutaneous mesenchymal stem cells: status of current knowledge,implications for dermatopathology

Stem cell biology is currently making its impact on medicine, which will probably increase over the next decades. It not only influences our therapeutic thinking caused by the enormous plasticity of stem cells but also affects diagnostic and conceptual aspects of dermatopathology. Although our knowledge of the keratinocytic stem cells located within the follicular bulge has exploded exponentially since their discovery in 1990, the concept of cutaneous mesenchymal stem cells (MSCs) is new. Described initially in 2001 in mice, MSCs later were also found in the human dermis. The connective tissue sheath and the papilla of the hair follicle probably represent the anatomical niche for cutaneous MSCs. In line with the cancer stem cell hypothesis, mutations of these cells may be the underlying basis of mesenchymal skin neoplasms, such as dermatofibrosarcoma protuberans. Furthermore, research on cutaneous MSCs may impact our thinking on the interaction of the epithelial component of skin neoplasms with their surrounding stroma. We are only in the early stages to recognize the importance of the potential contributions of cutaneous MSC research to dermatopathology, but it is not inconceivable to assume that they could be tremendous, paralleling the early discovery of the follicular bulge as a stem cell niche. Sellheyer K, Krahl D. Cutaneous mesenchymal stem cells: status of current knowledge, implications for dermatopathology.     

Skin‐derived mesenchymal stem cells (S‐MSCs) induce endothelial cell activation by paracrine mechanisms

Please cite this paper as: Skin‐derived mesenchymal stem cells (S‐MSCs) induce endothelial cell activation by paracrine mechanisms. Experimental Dermatology 2010; 19 : 848–850. Abstract: The mesenchymal stem cells (MSCs) are able to accumulate at the site of tissue damage. For this reason, they must transmigrate across the endothelium. In this study, we focused on skin‐derived MSCs (S‐MSCs), because the skin represents a useful stem cell source, and we analysed the VEGF released by S‐MSCs, because it is known to promote endothelial cell proliferation and vascular permeability. Moreover, we evaluated the influence of S‐MSC‐conditioned medium on human aortic endothelial cell intracellular calcium concentration ([Ca²⁺]_i) and nitric oxide (NO) production, given their important role in endothelial permeability modulation. Our results suggest that human S‐MSCs may interact with the endothelium via paracrine mechanisms, probably leading to an alteration of the endothelial barrier. Consequently, we could hypothesize that a therapeutic approach based on human skin‐derived MSCs may have a positive effect on tissue repair.     

Mesenchymal stem cell therapy in skin: why and what for?

In recent years, few stem cells have gained as much clinical notoriety as mesenchymal stem cells. Indeed, MSCs are already in use for a range of systemic inflammatory and autoimmune conditions that also affect the skin, such as acute and chronic graft versus host disease or lupus erythematosus. Most interestingly, these cells are able to improve skin wound healing in multiple preclinical models and few patient series. An additional potential of these cells is the delivery of missing structural elements in skin inherited disorders. However, we here contend that MSCs are not appropriate for cell replacement therapies in the context of wound healing. Indeed, engraftment of cells in the dermis is poor in the absence of irradiation and the observed effects seem mainly due to paracrine factors. In this viewpoint, we favour the hypothesis of a replete niche and competition with resident mesenchymal populations in the dermis not allowing the engraftment of newly delivered MSCs. Consequently, we propose that the benefit of MSCs may be at least in part reproduced by the growth factors or immunomodulatory molecules that they produce. In any case, the rapid progress in this field has allowed the emergence of important questions in skin biology that need to be addressed in parallel with the predictable future use of MSCs in the clinic.     

Perivascular localization of dermal stem cells in human scalp

In mammalian skin, the existence of stem cells in the dermis is still poorly understood. Previous studies have indicated that mesenchymal stem cells (MSCs) are situated as pericytes in various mammalian tissues. We speculated that the human adult dermis also contains MSC-like cells positive for CD34 at perivascular sites similar to adipose tissue. At first, stromal cells from adult scalp skin tissues showed colony-forming ability and differentiated into mesenchymal lineages (osteogenic, chondrogenic and adipogenic). Three-dimensional analysis of scalp skin with a confocal microscope clearly demonstrated that perivascular cells were positive for not only NG2, but also CD34, immunoreactivity. Perivascular CD34-positive cells were abundant around follicular portions. Furthermore, CD34-positive cell fractions collected with magnetic cell sorting were capable of differentiating into mesenchymal lineages. This study suggests that dermal perivascular sites act as a niche of MSCs in human scalp skin, which are easily accessible and useful in regenerative medicine.     

Tachykinins can partly explain the link within the neuroendocrine-immune-hematopoietic axis: novel role for mesenchymal stem cells

The tachykinins are among major regulators of bone marrow (BM) functions. The BM is resident to two stem cells: hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs). The adult BM has a finite number of HSCs that are required to replenish the immune system throughout life. MSCs surround BM vasculature, while HSCs are located close to the endosteal regions. The mechanisms by which the tachykinins regulate hematopoiesis require further research. Innervated fibers in the BM form synapse-like structures with MSCs. We propose that tachykinin-mediated effects on MSCs are relevant to hematopoiesis, because MSCs: 1) generate hematopoietic supporting stromal cells, 2) regulate the movement of cells in and out of the BM, and 3) regulate inflammatory responses in the BM. This study focuses on the truncated tachykinin receptor (NK-1-Tr). Its expression and regulation on MSCs mirrors the brain, but contrasts BM stroma. NK-1-Tr is predominantly expressed on MSCs. Because of limited receptor desensitization, we propose that NK-1-Tr on MSCs could allow rapid responses to the tachykinins so as to maintain the vascular/barrier functions of the BM, regulate immune responses to infectious agents that could be threat to BM failure, and respond to rapid loss of blood. Part of the mechanisms by which the tachykinins regulate MSCs functions involve novel interactions between IFN-γ and MHC class II. The findings described in this study hold clues to the role of tachykinins on hematopoiesis and adult stem cell functions in the BM. This report adds to an understanding of the crosstalk within the neural-immune-hematopoietic axis.     

The mesenchymal stem cell profile in psoriasis

Background The expression of inducible nitric oxide synthase (iNOS) and vascular endothelial growth factor (VEGF) and the level of total oxyradical scavenging capacity have been evaluated extensively in the cutaneous cells of patients with psoriasis. As yet, no indications are available about the undifferentiated cells, the mesenchymal stem cells (MSCs), isolated from skin. Objectives To isolate MSCs in patients with psoriasis and to compare them with those obtained from atopic and healthy subjects, in order to analyse whether MSCs show some typical psoriatic profiles and to understand whether pathophysiological events leading to psoriasis start early at the stem cell level. Methods MSCs isolated from seven patients with psoriasis, seven patients with acute atopic dermatitis and seven healthy subjects were characterized by fluorescence‐activated cell sorting analysis. VEGF and nitric oxide (NO) content was measured in conditioned medium, the expression of VEGF and iNOS was analysed by immunohistochemistry, and the total oxyradical scavenging capacity towards peroxynitrite was tested. Results VEGF content was highest in the medium conditioned by psoriatic perilesional MSCs, whereas NO concentration was maximally increased in medium conditioned by MSCs isolated from lesional psoriatic skin. The ability to neutralize the oxidizing effects of peroxynitrite was lower for MSCs isolated from lesional psoriatic skin compared with other MSCs, except for MSCs of lesional atopic skin. Conclusions The microenvironment in psoriasis differs from those of atopic dermatitis and healthy skin; it could induce resident MSCs to produce angiogenic and proinflammatory mediators which lead to a reduction in the antioxidant capacity of these cells, contributing to the development of skin lesions in psoriasis.     

Human skin-derived mesenchymal stem cells as a source of VEGF and nitric oxide

Researches on stem cells bring promise to functional skin repair. In particular, it has been recently suggested that mesenchymal stem cells (MSCs) could positively affect cutaneous wound healing through differentiation and paracrine action. The molecular mechanisms are not clear, even if there is increasing evidence for an important action of nitric oxide (NO), probably mediated by the regulation of the gene encoding for vascular endothelial growth factor (VEGF). The aim of our study was to investigate the immunohistochemical expression of VEGF and nitric oxide synthase (NOS) isoforms in human skin-derived MSCs, as well as the production of VEGF and NO, because these cells are less well characterized than bone marrow MSCs. MSCs were obtained from skin biopsies of healthy adult patients undergoing cosmetic plastic surgery, expanded and characterized for specific surface antigens. The cells were then evaluated for the immunohistochemical expression of VEGF, and NOS isoforms, as well as for VEGF and NO secretion in cell culture medium. Our immunohistochemical analysis showed that proliferating MSCs derived from human skin exhibit VEGF expression at cytoplasmic level as well as cytosolic and nuclear localization of all the three isoforms of NOS, even if with different patterns. In addition, our data evidenced the release of both VEGF and NO in cell culture supernatants. In conclusion, our results suggest that a therapeutic approach based on the human skin-derived MSCs may have a positive effect in wound healing conditions, through their ability to provide VEGF and NO to the damaged area.     

间充质干细胞在银屑病中的应用

【摘要】 间充质干细胞是干细胞家族的重要成员,具有自我更新、多向分化潜能和免疫调节作用及低免疫原性等特点。有文献报告间充质干细胞用于治疗银屑病获得不同程度的疗效,且有较长的疾病缓解期。本文综述间充质干细胞在银屑病中的应用及相关研究进展。