Exosome

摘要： Our previo us study demonstrated the potential therapeutic role of human neural stem cell-derived exosomes(hNSC-Exo)in ischemic stroke.Here,we loaded brain-derived neurotrophic factor(BDNF)into exosomes derived from NSCs to construct engineered exosomes(BDNF-hNSC-Exo)and compared their effects with those of hNSC-Exo on ischemic stroke both in vitro and in vivo.In a model of H_(2)O_(2)-induced oxidative stress in NSCs,BDNF-hNSC-Exo markedly enhanced cell survival.In a rat middle cerebral arte ry occlusion model,BDNF-hNSC-Exo not only inhibited the activation of microglia,but also promoted the differentiation of endogenous NSCs into neurons.These results suggest that BDNF can improve the function of NSC-derived exosomes in the treatment of ischemic stro ke.Our research may support the clinical use of other neurotrophic factors for central nervous system diseases.

摘要： Exosome-derived long non-coding RNAs(lncRNAs)are extensively engaged in recovery and repair of the injured spinal cord,through different mechanisms.However,to date no study has systematically evaluated the differentially expressed lncRNAs involved in the development of spinal cord injury.Thus,the aim of this study was to identify key circulating exosome-derived lncRNAs in a rat model of spinal cord injury and investigate their potential actions.To this end,we established a rat model of spinal cord hemisection.Circulating exosomes were extracted from blood samples from spinal cord injury and control(sham)rats and further identified through Western blotting and electron microscopy.RNA was isolated from the exosomes and sequenced.The enrichment analysis demonstrated that there were distinctively different lncRNA and mRNA expression patterns between the two groups.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis and Gene Ontology(GO)functional analysis were performed to determine the possible involvements of upregulated and downregulated lncRNAs in various pathways and different biological processes,as well as their cellular locations and molecular functions.Furthermore,quantitative reverse transcription-polymerase chain reaction showed that the expression of five lncRNAs––ENSRN0T00000067908,XR_590093,XR_591455,XR_360081,and XR_346933––was increased,whereas the expression of XR_351404,XR_591426,XR_353833,XR_590076,and XR_590719 was decreased.Of note,these 10 lncRNAs were at the center of the lncRNA-miRNA-mRNA coexpression network,which also included 198 mRNAs and 41 miRNAs.Taken together,our findings show that several circulating exosomal lncRNAs are differentially expressed after spinal cord injury,suggesting that they may be involved in spinal cord injury pathology and pathogenesis.These lncRNAs could potentially serve as targets for the clinical diagnosis and treatment of spinal cord injury.

摘要： Single-molecule detection and imaging are of great value in chemical analysis,biomarker identification and other trace detection fields.However,the localization and visualization of single molecule are still quite a challenge.Here,we report a special-engineered nanostructure of Ag nanoparticles embedded in multi-layer black phosphorus nanosheets(Ag/BP-NS)synthesized by a unique photoreduction method as a surfaceenhanced Raman scattering(SERS)sensor.Such a SERS substrate features the lowest detection limit of 10^(–20) mol L^(−1) for R6G,which is due to the three synergistic resonance enhancement of molecular resonance,photoinduced charge transfer resonance and electromagnetic resonance.We propose a polarization-mapping strategy to realize the detection and visualization of single molecule.In addition,combined with machine learning,Ag/BP-NS substrates are capable of recognition of different tumor exosomes,which is meaningful for monitoring and early warning of the cancer.This work provides a reliable strategy for the detection of single molecule and a potential candidate for the practical bio-application of SERS technology.

摘要： Exosomes derived from bone marrow mesenchymal stem cells can inhibit neuroinflammation through regulating microglial phenotypes and promoting nerve injury repair.However,the underlying molecular mechanism remains unclear.In this study,we investigated the mechanism by which exosomes derived from bone marrow mesenchymal stem cells inhibit neuroinflammation.Our in vitro co-culture experiments showed that bone marrow mesenchymal stem cells and their exosomes promoted the polarization of activated BV2 microglia to their anti-inflammatory phenotype,inhibited the expression of proinflammatory cytokines,and increased the expression of anti-inflammatory cytokines.Our in vivo experiments showed that tail vein injection of exosomes reduced cell apoptosis in cortical tissue of mouse models of traumatic brain injury,inhibited neuroinflammation,and promoted the transformation of microglia to the anti-inflammatory phenotype.We screened some microRNAs related to neuroinflammation using microRNA sequencing and found that microRNA-181b seemed to be actively involved in the process.Finally,we regulated the expression of miR181b in the brain tissue of mouse models of traumatic brain injury using lentiviral transfection.We found that miR181b overexpression effectively reduced apoptosis and neuroinflamatory response after traumatic brain injury and promoted the transformation of microglia to the anti-inflammatory phenotype.The interleukin 10/STAT3 pathway was activated during this process.These findings suggest that the inhibitory effects of exosomes derived from bone marrow mesenchymal stem cells on neuroinflamation after traumatic brain injury may be realized by the action of miR181b on the interleukin 10/STAT3 pathway.

摘要： Exosomes derived from mesenchymal stem cells are of therapeutic interest because of their important role in intracellular communication and biological regulation.On the basis of previously studied nerve conduits,we designed a polydopamine-modified chitin conduit loaded with mesenchymal stem cell-derived exosomes that release the exosomes in a sustained and stable manner.In vitro experiments revealed that rat mesenchymal stem cell-derived exosomes enhanced Schwann cell proliferation and secretion of neurotrophic and growth factors,increased the expression of Jun and Sox2 genes,decreased the expression of Mbp and Krox20 genes in Schwann cells,and reprogrammed Schwann cells to a repair phenotype.Furthermore,mesenchymal stem cell-derived exosomes promoted neurite growth of dorsal root ganglia.The polydopamine-modified chitin conduits loaded with mesenchymal stem cell-derived exosomes were used to bridge 2 mm rat sciatic nerve defects.Sustained release of exosomes greatly accelerated nerve healing and improved nerve function.These findings confirm that sustained release of mesenchymal stem cell-derived exosomes loaded into polydopamine-modified chitin conduits promotes the functional recovery of injured peripheral nerves.

摘要： Advances in regenerative medicine correlate strongly with progress in the use of adipose tissue-derived mesenchymal stem/stromal cells.The range of therapeutic indications has also expanded over recent years.Numerous recent studies have highlighted the primary importance of paracrine secretion by these cells.Though it is interesting to compare the different types of such secretions,we believe that exosomes(extra-cellular vesicles possessing the same properties as their source cells)will likely be the main key in tomorrow’s cell therapy.Exosomes also have many advantages compared to the direct use of cells,making these particles amajor target in fundamental and translational research.

摘要： Precision medicine is based on the identification of biomarkers of tumor development and progression.Liquid biopsy is at the forefront of the ability to gather diagnostic and prognostic information on tumors,as it can be noninvasively performed prior or during treatment.Liquid biopsy mostly utilizes circulating tumor cells,or free DNA,but also exosomes.The latter are nanovesicles secreted by most cell types,found in any body fluid that deliver proteins,nucleic acids and lipids to nearby and distant cells with a unique homing ability.Exosomes function in signalling between the tumor microenvironment and the rest of the body,promoting metastasis,immune remodelling and drug resistance.Exosomes are emerging as a key tool in precision medicine for cancer liquid biopsy,as they efficiently preserve their biomarker cargo.Moreover,exosomes strongly resemble the parental cell,which can help in assessing the oxidative and metabolic state of the donor cell.In this respect,exosomes represent one of the most promising new tools to fight cancer.This review will discuss the clinical applications of profiling exosomal proteins and lipids by high-throughput proteomics and metabolomics,and nucleic acids by next generation sequencing,as well as how this may allow cancer diagnosis,therapy response monitoring and recurrence detection.

摘要： Pain and lifestyle changes are common consequences of intervertebral disc degeneration(IVDD)and affect a large part of the aging population.The stemness of cells is exploited in the field of regenerative medicine as key to treat degenerative diseases.Transplanted cells however often face delivery and survival challenges,especially in tissues with a naturally harsh microniche environment such as the intervertebral disc.Recent interest in the secretome of stem cells,especially cargo protected from microniche-related decay as frequently present in degenerating tissues,provides new means of rejuvenating ailing cells and tissues.Exosomes,a type of extracellular vesicles with purposeful cargo gained particular interest in conveying stem cell related attributes of rejuvenation,which will be discussed here in the context of IVDD.

摘要： Mesenchymal stem cells(MSC)have pushed the field of stem cell-based therapies by inducing tissue regeneration,immunosuppression,and angiogenesis mainly through vesicles and soluble factors release(paracrine signaling).MSC-extracellular vesicles(MSC-EV)adaptable secretome and homing to injured sites allowed researchers to unlock a new era of cell-free based therapy.In parallel,nanoparticles(NP)have been explored in contributing to transport and drug delivery systems,giving drugs desired physical-chemical properties to exploit cell behavior.However,NPs can be quickly recognized by immune cells and cleared from circulation.In this viewpoint,we explore how combining both therapeutic strategies can improve efficacy and circumvent limitations of both therapies.MSCEV benefit from the potent MSC membrane composition,guiding chemotaxis to tumor sites,a very restricted microenvironment.MSC-EV has low immunogenicity,high stability,long half-life and can explore tissue targeting ligands as a precise drug carry,even across biological barriers.Those properties promote enhanced targeted drug delivery that can be combined with NP,exploring biological membrane production through:1.direct cell therapy with NP-infused MSC;2.NP-containing MSC-EV generated by NP-infused MSC;3.by coating NP in MSC membrane(“MSC NanoGhosts”),allowing precise cargo definition without losing targeting.Therefore,nanotechnology combined with cell-based therapeutic resources can greatly improve targeted drug delivery,improving efficacy and opening a new venue of therapeutic possibilities.

摘要： Stem cell research is a promising area of transplantation and regenerative medicine with tremendous potential for improving the clinical treatment and diagnostic options across a variety of conditions and enhancing understanding of human development.Over the past few decades,mesenchymal stem cell(MSCs)studies have exponentially increased with a promising outcome.However,regardless of the huge investment and the research attention given to stem cell research,FDA approval for clinical use is still lacking.Amid the challenges confronting stem cell research as a cellbased product,there appears to be evidence of superior effect and heightened potential success in its expressed vesicles,exosomes,as cell-free products.In addition to their highly desirable intrinsic biologically unique structural,compositional,and morphological characteristics,as well as predominant physiochemical stability and biocompatibility properties,exosomes can also be altered to enhance their therapeutic capability or diagnostic imaging potential via physical,chemical,and biological modification approaches.More importantly,the powerful therapeutic potential and superior biological functions of exosomes,particularly,regarding engineered exosomes as cell-free products,and their utilization in a new generation of nanomedicine treatment,vaccination,and diagnosis platforms,brings hope of a change in the near future.This viewpoint discusses the trend of stem cell research and why stem cell-derived exosomes could be the game-changer.

摘要： 本发明涉及一种离散汲取收集外泌体的方法及试剂，所述试剂包括PBS缓冲液、固定液及原水，所述方法包括：首先用一种抗凝溶剂将外泌体与DNA/RNA的大分子团分离，然后分级离心，除去DNA/RNA等大分子团，留存纯度较高的外泌体。其优点在于：(1)本发明提取外泌体的试剂可高效提取外泌体，收率高、纯度高。(2)本发明操作简单，成本低，对实验操作人员技术、实验设备要求低，有很好的应用前景，拓展了外泌体研究领域，对于外泌体的研究有重大意义。

摘要： 本发明涉及一种缓释iPSC‑MSC‑Exosomes温敏壳聚糖复合凝胶制备方法。技术方案是：缓释iPSC‑MSC‑Exosomes温敏壳聚糖复合凝胶制备方法，按以下步骤进行：1)将壳聚糖盐酸盐和明胶粉末溶于水中搅拌混匀；2)在步骤1)获得的溶液中逐滴加入0.444g/ml甘油磷酸二钠盐溶液，使得溶液中的壳聚糖：明胶：甘油磷酸二钠盐的最终比等于5:1.5‑2.5:7‑13，得到温敏壳聚糖凝胶溶液；3)在体外通过超速离心法提取iPSC‑MSC‑Exosomes；4)将iPSC‑MSC‑Exosomes逐渐加入温敏壳聚糖溶液中混合，得到温敏壳聚糖复合凝胶。该方法可持续释放外泌体以促进组织损伤修复。

摘要： 本发明提供了一种exosomes装载的miR‑93‑5p在治疗类风湿性关节炎中的应用，属于生物医学技术领域；本发明首先制备过表达或阻断miR‑93‑5p的G‑exosomes，并通过体外实验证明，过表达miR‑93‑5p的G‑exosomes更加明显的抑制了Th17细胞的分化；并通过研究过表达和阻断miR‑93‑5p的G‑exosomes对于CIA模型小鼠发病的作用，发现过表达miR‑93‑5p的G‑exosomes对CIA小鼠的保护作用更强了；而阻断miR‑93‑5p的G‑exosomes则没有上述作用；结果表明过表达miR‑93‑5p的G‑exosomes可用于类风湿性关节炎的治疗。

摘要： 本发明提供了一种携带抗肿瘤蛋白靶向性exosome的制备方法，包括以下步骤：(1)构建表达肿瘤靶向性多肽iRGD、exosome膜蛋白lamp2b及抗肿瘤蛋白PINT‑87aa的融合基因表达质粒；(2)将表达质粒转染到人的胚肾细胞HEK293细胞中，puro抗生素筛选获得稳定表达抗肿瘤融合蛋白的细胞系；(3)细胞扩增培养，对细胞释放出来的携带抗肿瘤蛋白的exosome进行分离纯化。本发明的方法成功构建了一种仿生物源性抗肿瘤蛋白输送载体，能够高效抑制肿瘤的生长，为恶性的治疗提供了一种潜在的理想的治疗方法。

摘要： 本发明涉及人脐带间质干细胞分泌的exosome治疗皮肤损伤的作用，具体涉及一种脐带间质干细胞分泌的直径为100nm左右的囊泡结构exosome在促进皮肤损伤修复中的药用作用，属于皮肤创伤治疗的技术领域；本发明所述的exosome是来源于人脐带间质干细胞，经试验验证，所涉及的exosome作为脐带间质干细胞细胞分泌的重要“成分”对皮肤损伤方面（包括烧烫伤和切除损伤）具有明显促修复的药用作用，而且其具有便于储存和运输的优势，这样脐带间质干细胞来源的exosome就为以间质干细胞为基础的非细胞治疗皮肤损伤提供了可能。

摘要： 本发明涉及一种NK细胞exosomes可控规模化的制备方法，包括以下步骤：(1)收集NK细胞培养基上清液，并用磷酸盐缓冲液进行稀释，得到稀释液；(2)将稀释液依次在低转速和高转速下离心，且每次离心后取上清液弃沉淀；(3)将离心上清液置于超滤管中进行离心，取上清液，得到的浓缩上清液进行超高速离心，超高速离心后取沉淀；(4)将步骤(3)超高速离心后得到的沉淀置于磷酸盐缓冲液中并转移至透析袋中进行透析，即得所述NK细胞exosomes。与现有技术相比，本发明粒径分布均匀，且表面携带标志性蛋白CD9、CD81、CD63，克服了现有制备技术存在纯度低，产量低，表征蛋白表达量低等不足，且可以直接用于后续的检测，安全系数高。

摘要： 本发明公开了一种尿液中近端肾小管来源exosome含量的检测方法，包括以下步骤：S1，制备尿液exosome悬液；S2，测定尿液exosome悬液中蛋白浓度；S3，分选尿液exosome悬液中近端肾小管来源exosome；S4，对S3中分选出的尿液中近端肾小管来源的exosome进行检测。本发明采用超速离心方法获得高纯度的尿液exosome，再采用免疫磁珠吸附exosome，克服了exosome体积小的问题，增加了流式细胞仪方法检测exosome的可行性；采用CD63和CD13免疫标记方法分选尿液中近端小管来源的exosome，并用于后续对尿液中近端肾小管来源exosome含量进行准确检测分析。

摘要： 本发明提供了一种肿瘤源性的exosome来源的与人类肝癌相关的miRNA标志物及其在诊断人类肝癌中的用途，所述miRNA该标志物为exosome来源的miRNA‑46a。本发明还提供了检测exosome来源的miRNA‑46a的试剂、含有该检测试剂的试剂盒及其在诊断人类肝癌中的用途。本发明还提供了用于检测exosome来源的miRNA‑46a的方法及其在诊断人类肝癌中的用途。本发明还提供了从少量血清中快速提取exosome的试剂、含有该exosome提取试剂的试剂盒以及使用该试剂盒从血液中提取exosome的方法。本发明的miRNA标志物及其检测试剂可用于肝癌的早期诊断，其灵敏性和特异性均优于现有技术。

摘要： 本发明提供了一种肿瘤源性的exosome来源的与人类肝癌相关的miRNA标志物及其在诊断人类肝癌中的用途，所述miRNA该标志物为exosome来源的miRNA‑33b。本发明还提供了检测exosome来源的miRNA‑33b的试剂、含有该检测试剂的试剂盒及其在诊断人类肝癌中的用途。本发明还提供了用于检测exosome来源的miRNA‑33b的方法及其在诊断人类肝癌中的用途。本发明还提供了从少量血清中快速提取exosome的试剂、含有该exosome提取试剂的试剂盒以及使用该试剂盒从血液中提取exosome的方法。本发明的miRNA标志物及其检测试剂可用于肝癌的早期诊断，其灵敏性和特异性均优于现有技术。

摘要： 本实用新型公开了一种用于体液样品exosome保存的试剂盒，包括盒体，所述盒体顶部活动连接有顶盖，所述顶盖与盒体之间设置有铰链，所述顶盖顶部外表面固定安装有把手，所述顶盖前壁固定安装有子扣，所述盒体前壁与子扣相对应位置固定安装有母扣，所述盒体内部设置有安装板，所述安装板内部设置有弹簧，所述弹簧一侧固定连接有第一短杆，所述第一短杆一侧末端固定连接有第一弧形夹板，所述安装板内壁中心轴线上固定连接有竖杆，本实用新型通过设置隔间、导热板及保温板，将冰块存放在隔间内部，避免打湿试剂盒底部，且导温板可以使得安装板内部温度保持在适合的温度左右，保温板也可以防止冰块过早的融化，该装置结构简单，方便实用。