作者感谢她的贡献:克里斯蒂安Pahrmann。特别感谢爱惜康，Norderstedt的，汉堡（德国）提供的缝合材料。 资金宋佳Schrepfer已收到来自德意志研究联合会（DFG）（SCHR992 / 3 1 SCHR992/4-1）研究补助金。这项工作是由ISHLT沙姆韦事业发展基金2010年和福尔克研究经费（斯坦福大学）的支持。

1。乳内动脉（IMA）准备<ol><li>动脉内皮剥脱2法国福格蒂动脉取栓导管（百特医疗用品，迪尔菲尔德，IL，USA）的通过。拉导管贯穿整个船舶长度的两倍，以确保内皮损伤。 </li><li>使用的长度为8mm和2.5毫米，直径3毫米（如Translumina，育空支架）的任何人的支架。注意:支架的直径应不超过10％以上的血管直径，以避免前，后支架狭窄。注意:请不要在同一研究中切换支架的长度。 </li><li>使用合适的球囊压力（这是在支架上包说明），以达到所需的直径部署支架。 </li><li>商店植入支架在4°C间的RPMI +肝素（500 IE/10毫升）上冰，直到移植IMA。 </li></ol> 2。动物的制备是侯，RNU裸体（CRL:NIH-Foxn1rnu）大鼠（300-350克）SED在scantainer常规条件下柜，标准大鼠饲料喂养和蒸压水广告libidum的的通风。 <ol><li>异氟醚麻醉大鼠（2.5-3％），使用的是感应室。 </li><li>剃腹毛，放置老鼠在它的后面，将超过它的鼻子和嘴，以保持麻醉面罩。 </li><li>消毒腹部地区广泛使用普罗沃碘，下次使用80％乙醇 - 重复此步骤两次。检查捏的后腿，要确保足够的麻醉大鼠的反射。 </li><li>在微观角度，执行上的中位数开腹暴露腹主动脉。 </li><li>将生理盐水滋润手套肠子。折叠肠子周围的手套，以防止水分流失。 </li><li>主动脉解剖，从下腹地区分叉，不小心引起​​血管的分支损伤。 </li><li>使用微观夹阻止主动脉的血流量。放置近端钳第一，其次是远端钳。 </li><li>删除一个应用程序。 0.5-0.7毫米的主动脉段和冲洗余下的主动脉与肝素（200单位）。 </li><li>采取的支架IMA和缩短到足够的长度和定位差距。 </li><li> IMA的连接到收件人主动脉，运行缝合使用8-0普理灵缝线（爱惜康，Norderstedt的，德国）。 </li><li>小心地打开第一颅钳，然后尾钳。 </li><li>应该有移植IMA与主动脉远端可见脉冲。 </li><li>将肠子放回腹部。 </li><li>预热的无菌生理盐水冲洗腹部。 </li><li>关闭腹部墙上使用6-0普理灵缝线（爱惜康，Norderstedt的，德国）的肌肉层。 </li><li>虽然仍然是在麻醉大鼠皮下注射，注射4-5毫克/公斤卡洛芬。 </li><li> 3天后外科手术后的镇痛管理适当（如卡洛芬或美洛昔康）红霉素。 </li></ol> 3。代表结果组织学标本固定在4％福尔马林，在酒精梯度脱水，1％干燥过氧化苯甲酰的80％和20％的甲基丙烯酸甲酯，邻苯二甲酸二丁酯为1天，我甲基丙烯酸甲酯的混合物（甲基丙烯酸甲酯一）渗透1天，和甲基丙烯酸甲酯我干3％的过氧化苯甲酰（三甲基丙烯酸甲酯）为1-2天，在4°C。此后，标本第三新鲜的甲基丙烯酸甲酯聚合在水浴上预聚合基地的玻璃小瓶。缓慢聚合取得小瓶保持在26°C过夜，提高温度至28°C的第二天早上，然后逐渐增加0.5°C，超过12小时的温度，直到聚合发生。聚合块切片在5微米厚度使用MICROM HM 360切片机，配备的钨硬质合金刀。 以确定细胞增殖的起源（图1），幻灯片staine确定绿色荧光蛋白（GFP）或鼠抗体ðMHC-Ⅰ和人类平滑肌细胞。对于这些研究，人类的IMA记者基因GFP过夜使用稳定的IMA细胞传导的慢病毒颗粒的培养。从人类起源的女儿细胞分裂可以识别表达GFP。经过deparaffinization，热诱导抗原检索进行加热的幻灯片，在抗原检索解决方案使用一个蒸笼。图像FX信号增强器可用于阻断步骤。人类起源的细胞确定对绿色荧光蛋白（在1:100稀释的一抗稀释液（DAKO））的小鼠单克隆抗体，并进一步的Alexa Fluor 488山羊抗鼠IgG（1:1000稀释在二次抗体稀释剂标记）。平滑肌细胞标记兔polycolonal抗平滑肌α-肌动蛋白（初级抗体稀释液稀释1:100），羊抗兔IgG的Alexa Fluor 555（1:1000 DIL在二次抗体稀释剂uted）。每个抗体孵育步骤进行1小时3次PBS洗涤之间，在37°C。细胞核用DAPI染色10分钟。安装使用延长黄金抗淬灭试剂幻灯片后，利用激光共聚焦显微镜样品进行了分析。 <img alt="图1" src="/files/ftp_upload/3663/3663fig1.jpg" /> 图1。作为人类平滑肌细胞内膜细胞。A:绿色=抗GFP的标记人类起源的细胞;红色=抗人平滑肌细胞肌动蛋白，蓝色的DAPI，确定细胞核乙 :绿色=抗大鼠的MHC-Ⅰ，贴标鼠源细胞;红色=抗人平滑肌细胞肌动蛋白，蓝色的DAPI，确定细胞的细胞核。被确定为平滑肌细胞和GFP阳性细胞增殖，但负鼠MHC-Ⅰ分子。因此，细胞增殖是人类的起源。

<ol>
	<li>Deuse, T., Ikeno, F., Robbins, R. C., Schrepfer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+In-Stent+Restenosis:+An+Inexpensive,+Reliable,+and+Rapid+Preclinical+Model.">Imaging In-Stent Restenosis: An Inexpensive, Reliable, and Rapid Preclinical Model.</a> J. Vis. Exp. (31), e1346 (2009).</li><li>Oyamada, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trans-iliac+rat+aorta+stenting:+a+novel+high+throughput+preclinical+stent+model+for+restenosis+and+thrombosis.">Trans-iliac rat aorta stenting: a novel high throughput preclinical stent model for restenosis and thrombosis.</a> J. Surg. Res. 166, e91-e95 (2011).</li><li>Chamberlain, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+mouse+model+of+in+situ+stenting.">A novel mouse model of in situ stenting.</a> Cardiovascular research. 85, 38-44 (2010).</li><li>Deuse, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Introducing+the+first+polymer-free+leflunomide+eluting+stent.">Introducing the first polymer-free leflunomide eluting stent.</a> Atherosclerosis. 200, 126-134 (2008).</li><li>Finn, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+healing+after+sirolimus,+paclitaxel,+and+bare+metal+stent+placement+in+combination+with+peroxisome+proliferator-activator+receptor+gamma+agonists:+requirement+for+mTOR/Akt2+in+PPARgamma+activation.">Differential healing after sirolimus, paclitaxel, and bare metal stent placement in combination with peroxisome proliferator-activator receptor gamma agonists: requirement for mTOR/Akt2 in PPARgamma activation.</a> Circulation research. 105, 1003-1012 (2009).</li><li>Tellez, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coronary+bare+metal+stent+implantation+in+homozygous+LDL+receptor+deficient+swine+induces+a+neointimal+formation+pattern+similar+to+humans.">Coronary bare metal stent implantation in homozygous LDL receptor deficient swine induces a neointimal formation pattern similar to humans.</a> Atherosclerosis. 213, 518-524 (2010).</li><li>Suzuki, Y., Yeung, A. C., Ikeno, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pre-clinical+animal+model+in+the+translational+research+of+interventional+cardiology.">The pre-clinical animal model in the translational research of interventional cardiology.</a> JACC Cardiovasc. Interv. 2, 373-383 (2009).</li><li>Holt, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intimal+proliferation+in+an+organ+culture+of+human+internal+mammary+artery.">Intimal proliferation in an organ culture of human internal mammary artery.</a> Cardiovascular research. 26, 1189-1194 (1992).</li><li>Swanson, N., Javed, Q., Hogrefe, K., Gershlick, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+internal+mammary+artery+organ+culture+model+of+coronary+stenting:+a+novel+investigation+of+smooth+muscle+cell+response+to+drug-eluting+stents.">Human internal mammary artery organ culture model of coronary stenting: a novel investigation of smooth muscle cell response to drug-eluting stents.</a> Clin. Sci. (Lond). 103, 347-353 (2002).</li></ol>

没有利益冲突的声明。

虽然在体内研究模型不同的是现有的调查支架植入后内膜增生的发展，这些车型仍然面临着转化的障碍需要克服。此外，大量的动物模型是昂贵和特殊的住房条件以及手术设备是不是所有实验室。 使用人类的IMA研究人类的内膜增生及支架内再狭窄的发展进行了研究之前， 当然在器官培养8,9 原位 。新的人性化的IMA模型构成了翻译的办法来解决支架...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> 2法国Fogarty导管</td><td>迪尔菲尔德，IL，美国百特医疗用品， </td><td> 120602F </td><td></td></tr><tr><td>育空支架</td><td> translumina有限公司，Hechingen，德国</td><td></td><td>根据您的研究方案，使用您所选择的支架</td></tr><tr><td>的RPMI媒体</td><td> biochrom </td><td> Nr.F1275 </td><td></td></tr><tr><td>肝素</td><td>巴克斯特</td><td> 2B0953 </td><td></td></tr><tr><td>异氟醚</td><td>住持</td><td> B506 </td><td></td></tr><tr><td>普罗沃碘</td><td>优碘Puredue医药</td><td>条形码:5995327165830 </td><td></td></tr><tr><td> 80％ethano升</td><td>赫耶尔</td><td>教育电视80/0500 </td><td></td></tr><tr><td>微型钳</td><td>哈佛仪器</td><td> PY2-61-0186 </td><td></td></tr><tr><td>缝线8-0 </td><td>强生公司</td><td> 2808G </td><td></td></tr><tr><td>缝线6-0 </td><td>强生公司</td><td> 1698Ĥ </td><td></td></tr><tr><td>卡洛芬</td><td> feizer兽医</td><td> PZN含量:0110208 </td><td></td></tr><tr><td>安乃近</td><td> Ratiopharm公司</td><td></td><td></td></tr><tr><td>目标检索解决方案，PH9 </td><td> Dako公司</td><td> S2368 </td><td></td></tr><tr><td>图像FX信号增强</td><td> Invitrogen公司</td><td> I36933 </td><td></td></tr><tr><td>鼠单克隆抗GFP抗体</td><td> BD的生活色彩</td><td> 632381 </td><td></td></tr><tr><td>初级抗体稀释剂</td><td> Dako公司</td><td> S3022 </td><td></td></tr><tr><td>的Alexa Fluor 488山羊抗鼠IgG </td><td> Invitrogen公司</td><td> A11017 </td><td></td></tr><tr><td>二次抗体稀释剂</td><td> Dako公司</td><td> S0809 </td><td></td></tr><tr><td>兔polycolonal抗平滑肌α-肌动蛋白</td><td> abcam， </td><td> ab5694 </td><td></td></tr><tr><td>的Alexa Fluor 555山羊抗兔IgG </td><td> Invitrogen公司</td><td> A21430 </td><td></td></tr><tr><td>延长黄金的抗淬灭剂</td><td> Invitrogen公司</td><td> P36930 </td><td></td></tr></tbody></table>

human internal mammary artery (ima) transplantation and stenting: a human model to study the development of in-stent restenosis

体内研究模型的临床前研究后内膜增生的血管内支架置入术的发展中的病理生物学和病理生理过程转化为1,2的方法是至关重要的。 常用的动物模型包括小鼠，大鼠，兔，猪3-5。然而，进入临床的设置，这些模型的翻译仍然困难，因为这些生物的进程已经开始研究在动物血管，但以前从未在人类研究模型6,7。在这部影片中，我们展示了一个新的人性化的模型，以克服这种转化的差距。所示的过程，重现性好，操作方便，快速执行，并适合学习内膜增生的发展和多样化支架的适用性。 该视频演示了如何执行，通过移植到免疫缺陷大鼠的血管支架技术，标识作为人类的起源细胞增殖。

Watch this Scientific Journal Video about Human Internal Mammary Artery IMA Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis at JoVE.com

Human Internal Mammary Artery IMA Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis

这个视频显示了一个模型来研究支架使用在人类免疫缺陷鼠模型的船只（IMA）的部署后内膜增生的发展。

人乳内动脉（IMA）移植及内支架置入术:人体模型支架内再狭窄的研究开发

Preclinical in vivo research models to investigate pathobiological and pathophysiological processes in the development of intimal hyperplasia after vessel ...

human-internal-mammary-artery-ima-transplantation-stenting-human

Research

JoVE Journal

Medicine

Human Internal Mammary Artery (IMA) Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis

11.2K 次观看.  TSI-Lab, Germany. 这个视频显示了一个模型来研究支架使用在人类免疫缺陷鼠模型的船只（IMA）的部署后内膜增生的发展。

视频: Human Internal Mammary Artery IMA Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

Utilizing Transcranial Magnetic Stimulation to Study the Human Neuromuscular System

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade. While the use of TMS has expanded to the study of many systems and processes during this time, the original application and perhaps one of the most common uses of TMS involves studying the physiology, plasticity and function of the human neuromuscular system. Single pulse TMS applied to the motor cortex excites pyramidal neurons transsynaptically 2 (Figure 1) and results in a measurable electromyographic response that can be used to study and evaluate the integrity and excitability of the corticospinal tract in humans 3. Additionally, recent advances in magnetic stimulation now allows for partitioning of cortical versus spinal excitability 4,5. For example, paired-pulse TMS can be used to assess intracortical facilitatory and inhibitory properties by combining a conditioning stimulus and a test stimulus at different interstimulus intervals 3,4,6-8. In this video article we will demonstrate the methodological and technical aspects of these techniques. Specifically, we will demonstrate single-pulse and paired-pulse TMS techniques as applied to the flexor carpi radialis (FCR) muscle as well as the erector spinae (ES) musculature. Our laboratory studies the FCR muscle as it is of interest to our research on the effects of wrist-hand cast immobilization on reduced muscle performance6,9, and we study the ES muscles due to these muscles clinical relevance as it relates to low back pain8. With this stated, we should note that TMS has been used to study many muscles of the hand, arm and legs, and should iterate that our demonstrations in the FCR and ES muscle groups are only selected examples of TMS being used to study the human neuromuscular system.

Transcranial magnetic stimulation (TMS) is a non-invasive tool to gain insight on the physiology and function of the human nervous system. Here, we present our TMS techniques to study cortical excitability of the upper limb and lumbar musculature.

利用经颅磁刺激，以研究人类的神经肌肉系统

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade.  While ...

Stem Cell Transplantation in an in vitro Simulated Ischemia/Reperfusion Model

Stem cell transplantation protocols are finding their way into clinical practice1,2,3. Getting better results, making the protocols more robust, and finding new sources for implantable cells are the focus of recent research4,5. Investigating the effectiveness of cell therapies is not an easy task and new tools are needed to investigate the mechanisms involved in the treatment process6. We designed an experimental protocol of ischemia/reperfusion in order to allow the observation of cellular connections and even subcellular mechanisms during ischemia/reperfusion injury and after stem cell transplantation and to evaluate the efficacy of cell therapy. H9c2 cardiomyoblast cells were placed onto cell culture plates7,8. Ischemia was simulated with 150 minutes in a glucose free medium with oxygen level below 0.5%. Then, normal media and oxygen levels were reintroduced to simulate reperfusion. After oxygen glucose deprivation, the damaged cells were treated with transplantation of labeled human bone marrow derived mesenchymal stem cells by adding them to the culture. Mesenchymal stem cells are preferred in clinical trials because they are easily accessible with minimal invasive surgery, easily expandable and autologous. After 24 hours of co-cultivation, cells were stained with calcein and ethidium-homodimer to differentiate between live and dead cells. This setup allowed us to investigate the intercellular connections using confocal fluorescent microscopy and to quantify the survival rate of postischemic cells by flow cytometry. Confocal microscopy showed the interactions of the two cell populations such as cell fusion and formation of intercellular nanotubes. Flow cytometry analysis revealed 3 clusters of damaged cells which can be plotted on a graph and analyzed statistically. These populations can be investigated separately and conclusions can be drawn on these data on the effectiveness of the simulated therapeutical approach.

Stem Cell Transplantation in an in vitro Simulated Ischemia/Reperfusion Model

We demonstrate how to set up an in vitro ischemia/reperfusion model and how to evaluate the effect of stem cell therapy on postischemic cardiac cells.

在干细胞移植在体外模拟缺血/再灌注模型

Stem cell transplantation protocols are finding their way into clinical practice1,2,3. Getting better results, making the protocols more robust, and ...

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human malignancies.1 Other than surgery for the minority of patients who present with localized disease, there is little or no survival benefit of systemic therapy. Therefore, there is a great need to better understand the biology of NETs, and in particular define new therapeutic targets for patients with nonresectable or metastatic neuroendocrine tumors. 3D cell culture is becoming a popular method for drug screening due to its relevance in modeling the in vivo tumor tissue organization and microenvironment.2,3 The 3D multicellular spheroids could provide valuable information in a more timely and less expensive manner than directly proceeding from 2D cell culture experiments to animal (murine) models.
To facilitate the discovery of new therapeutics for NET patients, we have developed an in vitro 3D multicellular spheroids model using the human NET cell lines. The NET cells are plated in a non-adhesive agarose-coated 24-well plate and incubated under physiological conditions (5% CO2, 37 &deg;C) with a very slow agitation for 16-24 hr after plating. The cells form multicellular spheroids starting on the 3rd or 4th day. The spheroids become more spherical by the 6th day, at which point the drug treatments are initiated. The efficacy of the drug treatments on the NET spheroids is monitored based on the morphology, shape and size of the spheroids with a phase-contrast light microscope. The size of the spheroids is estimated automatically using a custom-developed MATLAB program based on an active contour algorithm. Further, we demonstrate a simple method to process the HistoGel embedding on these 3D spheroids, allowing the use of standard histological and immunohistochemical techniques. 
This is the first report on generating 3D spheroids using NET cell lines to examine the effect of therapeutic drugs. We have also performed histology on these 3D spheroids, and displayed an example of a single drug's effect on growth and proliferation of the NET spheroids. Our results support that the NET spheroids are valuable for further studies of NET biology and drug development.

We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy.

人类神经内分泌肿瘤细胞株为研究人类神经内分泌肿瘤治疗的三维模型

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

颈动脉支架植入再狭窄的研究的小鼠模型

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic lesions. Mouse models are an important tool to investigate inflammatory processes in atherogenesis, but these models suffer from the phenotypic and functional differences between the murine and human immune system. In vitro cell experiments are used to specifically evaluate cell type-dependent changes caused by a substance of interest, but culture-dependent variations and the inability to analyze the influence of specific molecules in the context of the inflammatory compound in atherosclerotic lesions limit the impact of the results. In addition, measuring levels of a molecule of interest in human blood helps to further investigate its clinical relevance, but this represents systemic and not local inflammation. Therefore, we here describe a plaque culture model to study human atherosclerotic lesion biology ex vivo. In short, fresh plaques are obtained from patients undergoing endarterectomy or coronary artery bypass grafting and stored in RPMI medium on ice until usage. The specimens are cut into small pieces followed by random distribution into a 48-well plate, containing RPMI medium in addition to a substance of interest such as cytokines or chemokines alone or in combination for defined periods of time. After incubation, the plaque pieces can be shock frozen for mRNA isolation, embedded in Paraffin or OCT for immunohistochemistry staining or smashed and lysed for western blotting. Furthermore, cells may be isolated from the plaque for flow cytometry analysis. In addition, supernatants can be collected for protein measurement by ELISA. In conclusion, the presented ex vivo model opens the possibility to further study inflammatory lesional biology, which may result in identification of novel disease mechanisms and therapeutic targets.

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory process. This manuscript illustrates an easy to use ex vivo model to investigate fresh carotid or coronary artery plaques. The ex vivo model allows for the investigation of potential substances on the inflammatory milieu in human atherosclerotic lesions and results can be analyzed by various methods.

一个人力<em&gt;前体内</em&gt;动脉粥样硬化斑块模型研究病变的生物学

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic ...

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As such, primary or &#39;neuronal-like&#39; cell culture systems are commonly utilized. While&#160;these systems are relatively easy to work with, and are useful model systems in which various functional outcomes (such as cell death) can be readily quantified, the examined outcomes and pathways in cultured immature neurons (such as excitotoxicity-mediated cell death pathways) are not necessarily the same as those observed in mature brain, or in intact tissue. Therefore, there is the need to develop models in which cellular mechanisms in mature neural tissue can be examined. We have developed an in vitro technique that can be used to investigate a variety of molecular pathways in intact nervous tissue. The technique described herein utilizes rat cortical tissue, but this technique can be adapted to use tissue from a variety of species (such as mouse, rabbit, guinea pig, and chicken) or brain regions (for example, hippocampus, striatum, etc.). Additionally, a variety of stimulations/treatments can be used (for example, excitotoxic, administration of inhibitors, etc.). In conclusion, the brain slice model described herein can be used to examine a variety of molecular mechanisms involved in excitotoxicity-mediated brain injury.

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

We have developed a brain slice model which can be used to examine molecular mechanisms involved in excitotoxicity-mediated brain injury.&#160;This technique generates viable mature brain tissue and reduces animal numbers required for experimentation, whilst keeping the neuronal circuitry, cellular interactions, and postsynaptic compartments partly intact.

脑Microslices的兴奋性刺激作为<em&gt;在体外</em&gt;中风模型

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each year. While several animal models exist for this procedure and mice are the species that is most commonly used. The reasons for using mice are the relative cost of using this species, the existence of many genetically defined strains that allow for the study of immune responses, and the existence of an extensive array of reagents that can be used to further define responses in this species. This model has been used to define factors in the cornea that are responsible for the relative immune privilege status of this tissue that enables corneal allografts to survive acute rejection in the absence of immunosuppressive therapy. It has also been used to define those factors that are most important in rejection of such allografts. Consequently, much of what we know concerning mechanisms of both corneal allograft acceptance and rejection are due to studies using a murine model of corneal transplantation. In addition to describing a model for acute corneal allograft rejection, we also present for the first time a model of late-term corneal allograft rejection.

Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.

小鼠角膜移植模型，研究实体器官移植的最常见形式

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each ...

Establishment of a Human Multiple Myeloma Xenograft Model in the Chicken to Study Tumor Growth, Invasion and Angiogenesis

Multiple myeloma (MM), a malignant plasma cell disease, remains incurable and novel drugs are required to improve the prognosis of patients. Due to the lack of the bone microenvironment and auto/paracrine growth factors human MM cells are difficult to cultivate. Therefore, there is an urgent need to establish proper in vitro and in vivo culture systems to study the action of novel therapeutics on human MM cells. Here we present a model to grow human multiple myeloma cells in a complex 3D environment in vitro and in vivo. MM cell lines OPM-2 and RPMI-8226 were transfected to express the transgene GFP and were cultivated in the presence of human mesenchymal cells and collagen type-I matrix as three-dimensional spheroids. In addition, spheroids were grafted on the chorioallantoic membrane (CAM) of chicken embryos and tumor growth was monitored by stereo fluorescence microscopy. Both models allow the study of novel therapeutic drugs in a complex 3D environment and the quantification of the tumor cell mass after homogenization of grafts in a transgene-specific GFP-ELISA. Moreover, angiogenic responses of the host and invasion of tumor cells into the subjacent host tissue can be monitored daily by a stereo microscope and analyzed by immunohistochemical staining against human tumor cells (Ki-67, CD138, Vimentin) or host mural cells covering blood vessels (desmin/ASMA).
In conclusion, the onplant system allows studying MM cell growth and angiogenesis in a complex 3D environment and enables screening for novel therapeutic compounds targeting survival and proliferation of MM cells.

Human multiple myeloma (MM) cells require the supportive microenvironment of mesenchymal cells and extracellular matrix components for survival and proliferation. We established an in vivo chicken embryo model with engrafted human myeloma and mesenchymal cells to study effects of cancer drugs on tumor growth, invasion and angiogenesis.

建立一个人类多发性骨髓瘤异种移植模型中的鸡，研究肿瘤的生长，侵袭和血管生成

Multiple myeloma (MM), a malignant plasma cell disease, remains incurable and novel drugs are required to improve the prognosis of patients. Due to the ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular weight metabolites in tissues of the treated host to achieve long-term survival. The semipermeable membrane allows engrafted encapsulated cells to avoid rejection by the immune system. The encapsulation procedure was designed to enable a controlled release of bioactive compounds, such as insulin, other hormones, and cytokines. Here we describe a method for encapsulation of catabolic cells, which consume lipids for heat production and energy dissipation (thermogenesis) in the intra-abdominal adipose tissue of obese mice. Encapsulation of thermogenic catabolic cells may be potentially applicable to the prevention and treatment of obesity and type 2 diabetes. Another potential application of catabolic cells may include detoxification from alcohols or other toxic metabolites and environmental pollutants.

Here, we present a protocol for encapsulation of catabolic cells, which consume lipids for heat production in intra-abdominal adipose tissue and increase energy dissipation in obese mice.

封装产热前脂肪细胞移植到脂肪组织仓库

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

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Subtitles

人乳内动脉（IMA）移植及内支架置入术:人体模型支架内再狭窄的研究开发