差速贴壁法分离培养脂肪源间充质干细胞

目的:探讨差速贴壁法从大鼠腹股沟脂肪垫分离纯化脂肪源间充质干细胞(adipose tissue-derived mesenchymal stem cells,ADMSCs)的可行性。方法:采用差速贴壁培养法分离ADMSCs,并与普通培养法得到的ADMSCs进行表面分子CD44阳性率对比。在第2代ADMSCs中加入条件培养基进行诱导,根据条件培养基的不同分成3组:①成骨诱导组:加入成骨培养基;②成脂诱导组:加入成脂培养基;③对照组:仅加入基础培养基。成骨诱导组和对照组进行碱性磷酸酶(ALP)检测,成脂诱导组和对照组进行油红O染色检测。结果:差速贴壁培养法获得CD44阳性率更高的ADMSCs。成骨诱导组的ALP大大高于对照组,成脂诱导组油红O染色阳性,对照组油红O染色均为阴性。结论:差速贴壁培养法从大鼠腹股沟脂肪垫中分离得到高纯度ADMSCs。     

Inflammatory conditions affect gene expression and function of human adipose tissue-derived mesenchymal stem cells

There is emerging interest in the application of mesenchymal stem cells (MSC) for the prevention and treatment of autoimmune diseases, graft‐versus‐host disease and allograft rejection. It is, however, unknown how inflammatory conditions affect phenotype and function of MSC. Adipose tissue‐derived mesenchymal stem cells (ASC) were cultured with alloactivated peripheral blood mononuclear cells (PBMC) (mixed lymphocyte reaction: MLR), with proinflammatory cytokines [interferon (IFN)‐γ, tumour necrosis factor (TNF)‐α and interleukin (IL)‐6] or under control conditions, and their full genome expression and function examined. Proinflammatory cytokines mainly increased indoleamine‐2,3‐dioxygenase expression, whereas ASC cultured with MLR showed increased expression of COX‐2, involved in prostaglandin E₂ production. Both conditions had a stimulatory, but differential, effect on the expression of proinflammatory cytokines and chemokines, while the expression of fibrotic factors was decreased only in response to proinflammatory cytokines. Functional analysis demonstrated that inflammatory conditions affected morphology and proliferation of ASC, while their differentiation capacity and production of trophic factors was unaffected. The immunosuppressive capacity of ASC was enhanced strongly under inflammatory conditions. In conclusion, ASC showed enhanced immunosuppressive capacity under inflammatory conditions, while their differentiation capacity was preserved. Therefore, in vitro preconditioning provides ASC with improved properties for immediate clinical immune therapy.     

Isolation and characterization of exosomes from adipose tissue-derived mesenchymal stem cells

Mesenchymal stem cells (MSCs) are the subject of intense research as they are a potential therapeutic tool for several clinical applications. The new MSCs action models are focused on the use of MSC-derived secretome which contains several growth factors, cytokines, microRNAs, and extracellular vesicles such as exosomes. Exosomes have recently emerged as a component with great potential involved as mediators in cellular communication. The isolation and identification of exosomes has made it possible for them to be used in cell-free therapies. The purposes of this study are: (i) to detect exosomes released into adipose-derived MSC conditioned cell culture medium, (ii) to identify exosome morphology, and (iii) to carry out a complete characterization of said exosomes. Moreover, it is aimed at determining which method for exosome isolation would be best to use. Precipitation has been identified as a highly useful method of exosome isolation since it provides higher efficiency and purity values than other methods. A broad characterization of the exosomes present in the MSC-conditioned medium was also carried out. This work fills a gap in the existing literature on bioactive molecules which have attracted a great deal of interest due to their potential use in cellular therapies.     

成人脂肪源间充质干细胞在小鼠体内具有血液血管干细胞特性

目的:观察成人脂肪源Flk1+ CD31- CD34-细胞在小鼠体内是否具有血液血管干细胞的特性。方法:将男性成人脂肪源Flk1+ CD31- CD34-细胞经尾静脉输入6-8周龄、接受亚致死剂量射线照射的非肥胖型糖尿病及严重联合免疫缺陷(NOD/SCID)小鼠体内(每只输入剂量为105,悬于0.4m LRPMI-1640培养基中),对照组接受同等剂量的RPMI-1640培养基中。2个月后处死小鼠。用聚合酶链式反应、流式细胞术、三色荧光法及荧光原位杂交法检测脂肪源Flk1+ CD31- CD34-细胞在实验小鼠骨髓及消化系统内的分化情况。结果:脂肪源Flk1+ CD31- CD34-细胞在单细胞水平上在NOD/SCID小鼠体内可分化为内皮细胞和造血细胞。结论:脂肪源Flk1+ CD31- CD34-细胞在体内具有血液血管干细胞的特性,这类细胞有可能作为种子细胞用以治疗血液和血管性疾病。     

脂肪组织来源的干细胞的研究进展

来自中胚层的脂肪组织与骨髓组织一样含有大量能自我更新和多向系分化潜能的细胞群称为脂肪干细胞。其取材方便，来源丰富，可在体外稳定增殖传代。并与骨髓间充质干细胞有相似的多向分化表面标志CD105、STRO-1以及CD166受体。研究发现它具有多向系分化潜能，除可以分化为问充质来源的脂肪、骨，软骨、脂肪以及骨骼肌、心肌等细胞，也可诱导分化为来源于外胚层的神经细胞以及具有功能性的血管内皮细胞，用以修复骨、软骨、心肌、骨骼肌、血管以及神经等组织。其具有造血支持作用以及可被逆转录病毒、腺病毒以及慢病毒较高的转染效率等优点，可以作为基因转移良好的靶细胞。因而，脂肪来源的干细胞其有望成为组织工程、细胞治疗以及基因转染良好的种子细胞。     

间充质干细胞改善自身免疫性脑脊髓炎的作用研究

目的 探讨间充质干细胞(MSC)治疗自身免疫性脑脊髓炎(EAE)的机制.方法 用MOG35-55肽和弗氏完全佐剂乳化剂诱导建立C57BL/6小鼠的EAE模型;分离纯化培养骨髓来源MSC细胞;临床评分和脊髓组织切片评估小鼠的发病情况;酶联免疫吸附试验(ELISA)检测EAE组,MSC治疗组和对照组小鼠外周血细胞因子TNF-α,IFN-γ,IL-4和TGF-β的含量;流式细胞术分析3组小鼠脾脏细胞中CD4+ Foxp3+调节性T细胞(Treg)的比例变化.结果 分离纯化C57BL/6小鼠MSC成功;MSC治疗组小鼠的临床评分明显降低,且脊髓组织切片显示T细胞浸润显著减少;外周血中细胞因子IL-4,TGF-β显著高于对照组(t=7.719、17.17,P均＜0.01)和EAE组(t=54.45、48.36,P均＜0.01),IFN-γ,TNF-α低于对照组(t=104.90、1.998,P均＜0.01)和EAE组(t=270.1、13.58,P均＜0.01);脾脏细胞中CD4+ Foxp3+ Treg细胞的比例明显高于对照组(t=15.91,P＜0.01)和EAE组(t=33.39,P＜0.01).结论 MSC能够有效改善EAE小鼠的症状;其对EAE小鼠的治疗作用是通过上调抗炎细胞因子(IL-4,TGF-β)和Treg细胞并下调促炎因子(IFN-γ,TNF-α)的水平,来发挥免疫调节作用.     

脂肪间充质干细胞向髓核样细胞定向分化研究

目的探讨SD大鼠来源的髓核(nucleus pulposus,NP)细胞促使脂肪间充质干细胞(adipose tissue-derived mesenchymal stem cells,AMSCs)向NP样细胞定向分化的分子机制。方法采用酶消化法取脂肪细胞,极限稀释法纯化细胞;采用组织块培养法培养NP细胞。利用流式细胞技术,免疫荧光及RT-PCR检测对AMSCs及NP细胞进行鉴定。结果 AMSCs中Sca-1和CD44的阳性率较高,而CD45和CD11b阴性,共培养组荧光强度明显亮于单纯AMSCs组,AMSCs在NP细胞的诱导下聚焦蛋白聚糖(Aggrecan)、Ⅱ型胶原蛋白(CollagenⅡ)、Sox-9等表达水平较对照组高。结论共培养环境中髓核细胞分泌的可溶性因子TGF-1能促使AMSCs向NP样细胞定向分化。     

猪的脂肪和骨髓来源间充质干细胞培养特性比较

目的明确小型猪的脂肪来源间充质干细胞（A_r_MSCs）和猪的骨髓来源间充质干细胞（BM—MSCsl体外培养特性的异同。方法广西巴马小型猪,雌雄不限,猪龄4～6个月,体质量20～30kg。AT-MSCs来源于小型猪腹股沟皮下组织．BM—MSCs来源于小型猪的骨髓组织。培养AT-MSCs和BM—MSCs并观察它们的细胞形态。流式细胞仪检测Arr_MSCs和BM—MSCs的表面标志物（CD29、CD34、CD45、CD90）。分别观察Arr-MSCs和BM—MSCs的细胞生长分化能力;实时聚合酶链反应（PCR）检测基因表达。结果流式细胞仪检测结果表明,A．r_MSCs和BM—MSCs均表达CD29[分别为（99．06±0．30）％、（99．94±0．05）％]、CD90[分别为（97．404-0．40）％、（97．43±1J29）％1阳性,CD34、CD45阴性。AT-MSCs传代需培养5～7d,而BM—MSCs需培养7～10d。与BM—MSCs比较,AT—MSCs具有更强的生长分化能力。实时PCR检测基因表达结果显示．AT—MSCs和BM—MSCs均能分化心肌特异标志物a—skeletalactin和Troponin—I．二者差异无统计学意义．表明AT—MSCs和BM—MSCs均具备多项分化潜能．结论AT—MSCs是小型猪干细胞移植治疗的理想选择．     

脂肪间充质干细胞联合内皮祖细胞治疗小鼠溃疡性结肠炎

文题释义：间充质干细胞：是一种具有多分化潜能、可自我更新、低免疫原性的多能干细胞,可从脂肪、骨髓、脐带、胎盘等多种组织中分离。在体外培养时,间充质干细胞可在不同诱导条件下向多系细胞分化,移植到体内后可分化为相应器官、组织的实质和间质细胞。内皮祖细胞：ASAHARA等于1997年首次发现外周血中存在能分化为血管内皮细胞的前体细胞,将其命名为血管内皮祖细胞,在生理或病理因素刺激下,可从骨髓动员到外周血参与损伤血管的修复。在一定的诱导条件下还可向平滑肌细胞、心肌细胞等多种细胞分化。 摘要背景：已有研究表明间充质干细胞具有治疗溃疡性结肠炎的效果,内皮祖细胞对溃疡性结肠炎是否有治疗效果尚不明确,二者都是生物组织工程重要的种子细胞,均可进行移植。目的：探索脂肪间充质干细胞和内皮祖细胞联合移植治疗溃疡性结肠炎的效果。方法：C57BL/6J小鼠随机分为6组,每组24只：正常对照组、脂肪间充质干细胞组、内皮祖细胞组、联合移植组、糖皮质激素组、模型对照组。除正常对照组外,其他5组小鼠采用葡聚糖硫酸钠诱导建立溃疡性结肠炎模型。造模第7,10天各组小鼠分别经尾静脉注射脂肪间充质干细胞和/或内皮祖细胞、地塞米松磷酸钠、磷酸盐缓冲液。造模第12天处死小鼠,比较各组小鼠疾病活动指数、组织学病理评分、结肠长度、血清肿瘤坏死因子α水平等。结果与结论：①糖皮质激素组具有治疗溃疡性结肠炎的效果,与模型对照组比较差异有显著性意义(P < 0.05);②脂肪间充质干细胞组具有治疗溃疡性结肠炎的效果,优于糖皮质激素组,差异有显著性意义(P < 0.05);③内皮祖细胞组治疗溃疡性结肠炎的效果不显著,与模型对照组比较,差异无显著性意义(P > 0.05);④脂肪间充质干细胞联合内皮祖细胞移植治疗溃疡性结肠炎的效果优于其他各组,显著改善结肠缩短程度、疾病活动指数、组织学病理评分、血清肿瘤坏死因子α水平。 ORCID: 0000-0002-2409-0309(侯晓琳)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     

Adipose tissue-derived mesenchymal stem cells are more potent suppressors of dendritic cells differentiation compared to bone marrow-derived mesenchymal stem cells

Both mesenchymal stem cells (MSCs) and dendritic cells (DCs) are engaged in the regulation of the immune response parallel to their numerous functions.The main objective of this study was to compare the effects of mesenchymal stem cells isolated from human adipose tissue or human bone marrow on the expression of specific cell surface markers as well as the secretion of some cytokines by monocyte-derived dendritic cells. The set of methods used includes cell cultures, magnetic beads isolation of cells, flow cytometry, ELISA and proteome profiler kit assays. The results obtained show that MSCs isolated from human adipose tissue are more potent immunomodulators of differentiation of human DCs in comparison to the bone marrow-derived MSCs. In both cases the percentages of CD14+ cells were increased in co-cultures of MSCs and DCs and at the same time down-regulated the expression of CD80, CD86 and CD83 as in all experiments the effect of adipose tissue MSCs was stronger. Similarly, the secretion of IL-10 by dendritic cells was up-regulated in co-cultures of MSCs and dendritic cells and the effect was stronger when adipose tissue-derived MSCs were used.Taken together all results presented reveal the higher potential of the adipose tissue-derived MSCs to inhibit the differentiation and expression of functionally important co-stimulatory molecules on the surface of monocyte-derived dendritic cells than the bone marrow-derived MSCs.     

同种异体脂肪干细胞门脉移植对大鼠纤维化肝脏的影响

目的: 研究同种异体脂肪干细胞肝内移植对大鼠纤维化肝脏的影响。方法: 从SD大鼠腹膜脂肪组织提取脂肪干细胞,纯化、培养并稳定传代。雄性SD大鼠48只,成功建立大鼠肝纤维化模型。实验分为对照组(n=14)、门静脉组(n=11)和尾静脉组(n=14)。后两组分别经门静脉、尾静脉注射同种异体移植脂肪干细胞,观察移植前后大鼠肝血流动力学变化,并比较门静脉组和尾静脉组的疗效差异。结果: 成功提取脂肪干细胞,纯化、培养并稳定传代。输注脂肪干细胞前后各组肝脏计算机断层扫描灌注指数显示,门静脉组肝脏血流灌注有明显改善,且以门静脉灌注量为主(P<0.05),尾静脉组仅有轻微改善。组织学肝硬化评分显示,门静脉组见肝纤维化、肝细胞脂肪变性、肝细胞变性坏死均明显改善(P<0.05);尾静脉组仅肝细胞脂肪变性有改善。免疫组化示门静脉组肝脏微血管密度较其它组明显减少(P<0.05)。结论: 同种异体脂肪干细胞移植能改善肝纤维化SD大鼠的肝脏血流灌注及病理学改变;门静脉移植途径较尾静脉途径疗效更显著。     

Effects of expanded human adipose tissue-derived mesenchymal stem cells on the viability of cryopreserved fat grafts in the nude mouse

Adipose-derived mesenchymal stem cells (AdMSCs) augment the ability to contribute to microvascular remodeling in vivo and to modulate vascular stability in fresh fat grafts. Although cryopreserved adipose tissue is frequently used for soft tissue augmentation, the viability of the fat graft is poor. The effects of culture-expanded human adipose tissue-derived mesenchymal stem cells (hAdMSCs) on the survival and quality of the cryopreserved fat graft were determined. hAdMSCs from the same donor were mixed with fat tissues cryopreserved at -70 °C for 8 weeks and injected subcutaneously into 6-week-old BALB/c-nu nude mice. Graft volume and weight were measured, and histology was evaluated 4 and 15 weeks post-transplantation. The hAdMSC-treated group showed significantly enhanced graft volume and weight. The histological evaluation demonstrated significantly better fat cell integrity compared with the vehicle-treated control 4 weeks post-transplantation. No significant difference in graft weight, volume, or histological parameters was found among the groups 15 weeks post-transplantation. The hAdMSCs enhanced the survival and quality of transplanted cryopreserved fat tissues. Cultured and expanded hAdMSCs have reconstructive capacity in cryopreserved fat grafting by increasing the number of stem cells.     

Influence of mechanical fluid shear stress on the osteogenic differentiation protocols for Equine adipose tissue-derived mesenchymal stem cells

Cell-based therapies have become a promising approach to promote tissue regeneration and the treatment of musculoskeletal disorders. Bone regeneration maintains bone homeostasis, mechanical stability and physical performance. Mechanical stimulation showed to induce stem cell differentiation into the osteogenic fate. However, the effect of various osteogenic protocols on the osteogenic commitment of equine adipose-derived stem cells is not fully elucidated. Here we examined the influence of fluid-based shear stress (FSS) via mechanical rocking to assess whether mechanical stimulation promotes osteogenic differentiation of equine adipose-derived stem cells (ASCs). ASCs were induced into osteogenic fate using osteogenic differentiation medium (ODM) protocol or additional supplementation of 5?mM CaCl₂ and 7.5?mM CaCl₂ protocol compared to cells cultivated in basal medium (BM) up to 21 day. The ASCs proliferation pattern was evaluated using the sulforhodamine B (SRB) protein assay. Osteogenic differentiation examined via semi-quantification of alizarin red staining (ARS) and alkaline phosphatase activity (ALP) as well as, via quantification of osteocalcin (OC), alkaline phosphatase (ALP), osteopontin (OP), and collagen type-1 (COL1) gene expression using RT-qPCR. We show that mechanical FSS increased the proliferation pattern of ASCs compared to the static conditions. Mechanical FSS together with 5?mM CaCl₂ and 7.5?mM CaCl₂ promoted osteogenic nodule formation and increased ARS intensity compared to the standard osteogenic protocols. We observed that combined mechanical FSS with ODM protocol increase ALP activity compared to static culture conditions. We report that ALP and OC osteogenic markers expression were upregulated under mechanical FSS culture condition particularly with the ODM protocol. Taken together, it can be assumed that mechanical stress using FSS promotes the efficiency of the osteogenic differentiation protocols of ASCs through independent mechanisms.     

间充质干细胞在自身免疫性疾病中的应用研究进展

自身免疫性疾病（AID）的发病机制主要在于机体自身耐受的破坏,机体产生自身抗体和（或）自身反应性淋巴细胞,导致疾病的发生。目前临床上对AID的治疗主要是采取非特异性免疫抑制。虽然一定程度上可减轻症状,但不能根治疾病,而重症AID缺乏理想的治疗方法,预后差。因此,寻找有效的治疗方法仍然是目前临床亟待解决的问题。间充质干细胞（MSC）是一种非造血多能成体干细胞,具有多向分化以及促进组织修复等潜能,其免疫调控作用的发现是近年来干细胞研究领域的一项重要突破。最近,MSC移植治疗自身免疫性疾病的应用研究不断涌现,显示了MSC的免疫调节特性及其治疗AID的潜在能力,为进一步研究奠定了基础。     

脂肪源间充质干细胞治疗aGVHD分子机制的初步研究

目的初步探讨成人脂肪源间充质干细胞（adult adipose tissue-derived mesenchymal stem cells．AMSC）治疗急性移植物抗宿主病（acute graft-versus-host disease，aGVHD）的分子机制。方法3例行异基因造血干细胞移植术后发生aGVHD患者，以每公斤体质量2×10^6个细胞剂量静脉输注AMSC；应用RT-PCR扩增患者AMSC使用前后外周血单个核细胞的TCR Vβ 24个亚家族的CDR3，了解患者TCR Vβ T细胞的分布情况；应用尼龙毛柱分离外周血T淋巴细胞，再经CD8磁珠分选出CD8^＋ T淋巴细胞．应用流式细胞术检测发生aGVHD患者使用AMSC前后外周血CD8^＋T细胞亚群的变化。结果患者发生aGVHD时，有TCR Vβ3及其他亚家族基因表达，输注AMSC后，GVHD得以有效控制。Vβ3不表达；当患者GVHD复发时，Vβ3基因又表达，治疗后不表达；与输注AMSC前相比，输注AMSC后，CD8^＋T细胞中的CD8^＋CD28^-亚群显著上调（P〈0．05），同时．患者的aGVHD得以有效控制。结论AMSC治疗aGVHD的分子机制可能与其抑制TCR Vβ亚家族基因表达有关，TCR Vβ3可能是AMSC作用的靶基因；同时，AMSC治疗aGVHD的作用机制可能与其上调CD8^＋CD28^-T细胞亚群有关．CD8^＋T细胞可能是AMSC作用的靶细胞。     

晚期糖基化终产物对人脂肪间充质干细胞功能影响的体外研究*

 目的：探讨晚期糖基化终产物（AGEs）对人脂肪来源的间充质干细胞（hADSCs）促进创伤修复功能的影响。方法：体外培养hADSCs,实验分为牛血清白蛋白（BSA）对照组、低浓度糖基化修饰的牛血清白蛋白（AGE-BSA）效应组和高浓度AGE-BSA效应组。采用WST法和Transwell迁移实验检测各组细胞增殖和迁移情况。应用实时定量PCR和ELISA检测各组细胞分泌血管内皮生长因子（VEGF）、肝细胞生长因子（HGF）和胰岛素样生长因子1（IGF-1）的表达。结果：与对照组相比, AGE-BSA组增殖能力和迁移能力明显下降（P<0.05）,VEGF、HGF和IGF-1 mRNA及蛋白表达水平显著降低（P<0.05）。结论：AGEs能损伤hADSCs的促进创伤修复功能,因而能够影响hADSCs治疗糖尿病皮肤溃疡病的治疗效果。     

Collagen I gel promotes homogenous osteogenic differentiation of adipose tissue-derived mesenchymal stem cells in serum-derived albumin scaffold

Repair of bone defects is a difficult clinical problem for reconstructive surgeons. Bone tissue engineering using an appropriate scaffold with cells is a new therapy for the repair of bone defects. The aim of this study was to evaluate the in vitro osteogenesis of canine adipose tissue-derived mesenchymal stem cells (Ad-MSCs) cultured in a combination of collagen I gel and a porous serum-derived albumin scaffold. A serum-derived albumin scaffold was prepared with canine serum by cross-linking and freeze-drying procedures. Ad-MSCs were seeded into serum-derived albumin scaffolds with or without collagen I gel, and were exposed to osteogenic differentiation conditions in vitro. After 28?days of in vitro culture, the distribution and osteogenic differentiation of Ad-MSCs cultured in the scaffold were evaluated by scanning electron microscopy, histology, immunohistochemistry, alkaline phosphatase (ALP) activity assay, and calcium colorimetric assay. Ad-MSCs showed more homogeneous distribution and osteogenic differentiation in the scaffold with collagen I gel than without collagen I gel. ALP activity and extracellular matrix mineralization in the construct with type I collagen were significantly higher than in the construct without type I collagen (p?<?0.05). In conclusion, the combination of collagen I gel and the serum-derived albumin scaffold enhanced osteogenic differentiation and homogenous distribution of Ad-MSCs.     

骨髓间充质干细胞对致敏小鼠造血干/祖细胞移植植入的影响

目的: 前期的研究已经证实致敏小鼠造血干/祖细胞移植植入失败率高。本研究拟通过骨髓间充质干细胞(MSCs)进行干预,观察能否提高造血干、祖细胞移植的植入率。方法: 应用贴壁培养法体外培养正常小鼠骨髓MSCs,并分为6个实验组,包括实验组1:d11 MSCs干预的致敏组;实验组2: d0 MSCs干预的致敏组;实验组3:d11和d0 2次MSCs干预的致敏组;实验组4: 无MSCs干预的致敏小鼠对照组;实验组5:无MSCs干预的正常小鼠(非致敏小鼠)移植对照组;实验组6:无MSCs干预的正常小鼠不移植对照组。观察指标包括生存分析、移植效果分析(血象改变、骨髓细胞恢复及嵌合分析等)和移植物抗宿主病(GVHD)检测,最终评估MSCs干预对各实验组异基因造血干/祖细胞移植植入率的影响效果。结果: 与对照组(实验组4、5、6)比较,MSCs干预(实验组1、2、3)在2次异基因脾细胞注射法致敏的动物模型进行异基因造血干/祖细胞移植时,未能促进骨髓造血干/祖细胞移植的植入,也未能延长致敏动物移植后的生存时间。结论: 体内应用1×10⁶ MSCs干预,未能促进2次异基因1×10⁶ C57BL/6小鼠脾细胞输注法建立的重度致敏模型异基因造血干/祖细胞移植的植入。     

The effect of mesenchymal stem cells on dynamic changes of T cell subsets in experimental autoimmune uveoretinitis

Mesenchymal stem cells (MSCs) are being explored extensively as a promising treatment for autoimmune diseases. We have recently reported that MSCs could ameliorate experimental autoimmune uveoretinitis (EAU) in rats. In this study, we examined further the effects of MSCs on the dynamics of T cell subsets in both eye and spleen and their cytokine production during the course of EAU. We focused on when and where the MSCs had inhibitory effects on T helper type 1 (Th1) and Th17 cells and how long the inhibitory effect lasted, in order to provide more mechanistic evidence for MSCs on the treatment of uveitis. Compared to the control group, administration of MSCs decreased the production of Th1 and Th17 cytokines significantly, while the production of Th2 and regulatory T cell (T_reg) cytokines [interleukin (IL)‐10 and transforming growth factor (TGF)‐β] was elevated during the entire course of EAU. Correspondingly, the dynamic levels of IL‐17 in the aqueous humour (AqH) were reduced in MSC‐treated rats. Moreover, the ratio of Th17/T_reg cells in both spleen and eye was decreased. These results provide powerful evidence that MSCs can regulate negatively both Th1 and Th17 responses and restore the balance of Th17/T_regs in the whole course of EAU, which is important for the regression of the disease.