Name: engineering biological-based vascular grafts using a pulsatile bioreactor
Uploaded: 2011-06-14T12:00:00
Description: Watch this Scientific Journal Video about Engineering Biological-Based Vascular Grafts Using a Pulsatile Bioreactor at JoVE.com

这项工作是由美国国立卫生批准的EB - 008836 R01，R01 HL083895（包括到LEN）。我们要感谢为我们研究的生物反应器，大学吹玻璃，达瑞尔史密斯。

高压灭菌器组装和高压灭菌管流系统和生物反应器组件（生物反应器本身和硅胶塞盖），在图1和图2的指示。鼻饲管，有一个男性一端连接器和一个对方开放。三是通过气体交换的硅胶帽插入短管段。 1。缝纫机的PGA网<ol><li>剪下的PGA网格1.1厘米X〜8厘米表（依赖于生物反应器的大小）。 </li><li>用蒸馏水（dH20）和空气干燥，使用前清洁硅胶管（3毫米内径）...

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	<li>Risau, W., Flamme, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vasculogenesis.">Vasculogenesis.</a> Annu. Rev. Cell Dev. Biol. 11, 73-91 (1995).</li><li>Fankhauser, F., Bebie, H., Kwasniewska, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Influcence+of+mechanical+Forices+and+Flow+Mechanisms+on+Vessel+Occlusion.">The Influcence of mechanical Forices and Flow Mechanisms on Vessel Occlusion.</a> Lasers in Surgery and Medicine. 6, 530-532 (1987).</li><li>Niklason, L. E., Gao, J., Abbott, W. M., Hirschi, K. K., Houser, S., Marini, R., Langer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+arteries+grown+in+vitro.">Functional arteries grown in vitro.</a> Science. 284, 489-493 (1999).</li><li>Prabhakar, V., Grinstaff, M. W., Alarcon, J., Knors, C., Solan, A. K., Niklason, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+porcine+arteries:+Effects+of+scaffold+modification.">Engineering porcine arteries: Effects of scaffold modification.</a> Journal of Biomedical Materials Research Part A. 67A, 303-311 (2003).</li><li>Mitchell, S. L., Niklason, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Requirements+for+growing+tissue-engineered+vascular+grafts.">Requirements for growing tissue-engineered vascular grafts.</a> Cardiovascular Pathology. 12, 59-64 (2003).</li><li>Dahl, S. L. M., Rhim, C., Song, Y. C., Niklason, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+properties+and+compositions+of+tissue+engineered+and+native+arteries.">Mechanical properties and compositions of tissue engineered and native arteries.</a> Annals of Biomedical Engineering. 35, 348-355 (2007).</li><li>Quint, C., Kondo, Y., Manson, R. J., Lawson, J. H., Dardik, A., Niklason, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decellularized+tissue-engineered+blood+vessel+as+an+arterial+conduit.">Decellularized tissue-engineered blood vessel as an arterial conduit.</a> Proc Natl Acad Sci U S A. 108, 9214-9219 (2011).</li><li>Niklason, L. E., Yeh, A. T., Calle, E. A., Bai, Y., Valent&#237;n, A., Humphrey, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enabling+Tools+for+Engineering+Collagenous+Tissues+Integrating+Bioreactors,+Intravital+Imaging,+and+Biomechanical+Modeling.">Enabling Tools for Engineering Collagenous Tissues Integrating Bioreactors, Intravital Imaging, and Biomechanical Modeling.</a> Proceedings of the National Academy of Sciences of the United States of America. 107, 3335-3339 (2010).</li><li>Gong, Z., Calkins, G., Cheng, E. -. c., Krause, D., Niklason, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+Culture+Medium+on+Smooth+Muscle+Cell+Differentiation+from+Human+Bone+Marrow-Derived+Mesenchymal+Stem+Cells.">Influence of Culture Medium on Smooth Muscle Cell Differentiation from Human Bone Marrow-Derived Mesenchymal Stem Cells.</a> Tissue Engineering Part A. 15, 319-330 (2009).</li><li>Gong, Z. D., Niklason, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small-diameter+human+vessel+wall+engineered+from+bone+marrow-derived+mesenchymal+stem+cells+(hMSCs.">Small-diameter human vessel wall engineered from bone marrow-derived mesenchymal stem cells (hMSCs.</a> Faseb Journal. 22, 1635-1648 (2008).</li><li>Poh, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+vessels+engineered+from+human+cells.">Blood vessels engineered from human cells.</a> Lancet. 365, 2122-2124 (2005).</li><li>American Heart Association. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Biostatistical fact sheet: cardiovascular procedures. , (2002).</li></ol>

工程船的质量是在组织培养中使用的校董会的质量决定的很大一部分。 SMC的表型的一个关键方面，包括收缩形态，传代次数低​​，和生物反应器内的能力。我们建议，通过在细胞种植的时间比P3的更大的聚合物支架上。此外，关键是要确认，SMC源是支原体免费使用前。我们观察到，支原体污染的细胞在生物反应器培养细胞增殖和胶原基质沉积的大幅下跌导致。 40多万冠状?...

<table border="1"><tbody><tr><th>试剂/材料名称</th><th>供应商</th><th>目录编号</th></tr><tr><td>胎牛血清（胎牛血清）热灭活</td><td> Hyclone公司</td><td> SH30071 </td></tr><tr><td> DMEM培养液</td><td> GIBCO公司</td><td> 11885 </td></tr><tr><td> rhFGF的基本</td><td> R＆B </td><td> 234 - FSE </td></tr><tr><td> rrPDGF - BB </td><td> R＆B </td><td> 520 - BB </td></tr><tr><td>青霉素摹</td><td>西格玛</td><td>彭纳</td></tr><tr><td>铜（II）硫酸</td><td>西格玛</td><td> C8027 </td></tr><tr><td> Gylcine </td><td>西格玛</td><td> C8790 </td></tr><tr><td> L -丙氨酸</td><td>西格玛</td><td> A7469 - 25G </td></tr><tr><td> L -脯氨酸</td><td>西格玛</td><td> P5607 - 25G </td></tr><tr><td>抗坏血酸的</td><td>西格玛</td><td> A4544 - 25G </td></tr><tr><td> HEPES </td><td>西格玛</td><td> H3375 - 100G </td></tr><tr><td>硅胶塞</td><td>科尔 - 帕默</td><td> 06298-24 </td></tr><tr><td>的Masterflex管的L / S </td><td>科尔 - 帕默</td><td> 06508-16，06508-18 </td></tr><tr><td>的Masterflex泵</td><td>科尔 - 帕默</td><td> 7553-80 </td></tr><tr><td>涤纶袖</td><td> MAQUET </td><td> 174406 </td></tr><tr><td> PGA的感觉</td><td>康科迪亚</td><td> MO000877 - 01 </td></tr><tr><td> 4-0 1.5公吨Surgipro第二缝合</td><td> Syneture </td><td> VP - 557 - X </td></tr><tr><td> 6-0 0.7公吨敌克松缝合</td><td> Syneture </td><td> 7538-11 </td></tr><tr><td> 0.22μm的聚四氟乙烯滤波器</td><td>瓦特曼</td><td> 6780-2502 </td></tr><tr><td>三通停止公鸡</td><td>爱德华生命科学</td><td> 593WSC </td></tr><tr><td>压力传感器</td><td>爱德华生命科学</td><td> PX212 </td></tr><tr><td>输液袋</td><td>巴克斯特</td><td> R4R2110 </td></tr><tr><td>盐水稀释集</td><td>箭头</td><td> W20030 </td></tr><tr><td>硅胶管</td><td>圣戈班</td><td> F05027 </td></tr></tbody></table>

engineering biological-based vascular grafts using a pulsatile bioreactor

一直致力于发展和推进的方法再生功能的小直径动脉绕过许多努力。在生理环境，机械和化学刺激，需要保持适当的发展和1,2动脉血管的功能。 由本集团开发的生物反应器文化系统的设计，支持范围内的本地船只的精确控制化学机械模仿环境的血管再生。我们的生物反应器的组装和维修程序相当简单，高度可重复的 3,4的。平滑肌细胞（校董会）接种于管状聚乙醇酸（PGA）目，是超过兼容的硅胶管螺纹，带或不带长达12周的搏动刺激和生物反应器中培养。区别于一些前辈，我们的生物反应器有四个主要属性。 1）不同于其他文化系统，模拟生化本地血管周围，我们的生物反应器也创造文化的船只申请环径向应变生理搏动环境。 2）多个工程船只可以同时内控制化学环境不同的机械条件下培养。 3）生物反应器，允许单内皮细胞层（EC）可以很容易地涂到动物植入模型工程血管的管腔侧。 4）我们的生物反应器还可以具有不同直径大小的文化工程的船只从1毫米至3毫米不等，节约的努力调整每一个人的生物反应器，以适应特定的直径大小。 在我们的生物反应器培养工程船类似于原生的血管组织在一定程度上。在血管壁上的细胞表达成熟的SMC的收缩标记，如平滑肌肌球蛋白重链（SMMHC）3 。大量的胶原沉积在细胞外基质，这是负责的5个工程船只的最终机械强度。生化分析还表明，胶原蛋白的工程船只的内容是与 6个原生动脉。重要的是，脉冲式生物反应器一直再生的船只，表现出机械性能，允许在动物模型的成功植入实验3,7。此外，这种生物反应器，可以进一步修改，以允许实时跟踪和评估胶原重构随着时间的推移，非侵入性，使用一个非线性光学显微镜（NLOM）8。最后，这种生物反应器应该作为一个很好的平台，究其根本机制，规范再生功能的小口径血管移植。

Watch this Scientific Journal Video about Engineering Biological-Based Vascular Grafts Using a Pulsatile Bioreactor at JoVE.com

Engineering Biological-Based Vascular Grafts Using a Pulsatile Bioreactor

本集团已开发出一种生物反应器的文化系统，模仿对心血管系统的生理搏动强调，重新植入式小直径血管移植。

基于生物工程，人造血管使用脉动生物反应器

Much effort has been devoted to develop and advance the methodology to regenerate functional small-diameter arterial bypasses. In the physiological ...

Research

JoVE Journal

Bioengineering

Design of a Biaxial Mechanical Loading Bioreactor for Tissue Engineering

组织工程生物反应器的双轴机械装载设计

We  designed a loading device that is capable of applying uniaxial or biaxial  mechanical strain to a tissue engineered biocomposites fabricated for  ...

科研

生物工程

Operation of a Benchtop Bioreactor

一个台式生物反应器的操作

Fermentation systems are used to provide an optimal growth environment for many different types of cell cultures. The ability afforded by fermentors to ...

Nonhuman Primate Lung Decellularization and Recellularization Using a Specialized Large-organ Bioreactor

非人灵长类龙脱细胞和Recellularization使用专门的大型器官生物反应器

There are an insufficient number of lungs available to meet current and future organ transplantation needs. Bioartificial tissue regeneration is an ...

Construction and Characterization of a Novel Vocal Fold Bioreactor

一种新型声带生物反应器的构建与鉴定

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To ...

Cultivation of Mammalian Cells Using a Single-use Pneumatic Bioreactor System

培养哺乳动物细胞使用一次性使用气动生物反应器系统

Recent advances in mammalian, insect, and stem cell cultivation and scale-up have created tremendous opportunities for new therapeutics and personalized ...

Surgical Technique for the Implantation of Tissue Engineered Vascular Grafts and Subsequent In Vivo Monitoring

Surgical Technique for the Implantation of Tissue Engineered Vascular Grafts and Subsequent In Vivo Monitoring

手术技术组织工程血管移植和后续的植入<em&gt;在体内</em&gt;监控

The development of Tissue Engineered Vessels (TEVs) is advanced by the ability to routinely and effectively implant TEVs (4-5 mm in diameter) into a large ...

Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration

工程三维细胞化胶原凝胶的血管组织再生

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. ...

A Novel Bioreactor for High Density Cultivation of Diverse Microbial Communities

一种新型生物反应器为不同的微生物群落的高密度栽培

A novel reactor design, coined a high density bioreactor (HDBR), is presented for the cultivation and study of high density microbial communities. Past ...

Image-guided, Laser-based Fabrication of Vascular-derived Microfluidic Networks

影像引导下，血管源性微流控网络的基于激光的研制

This detailed protocol outlines the implementation of image-guided, laser-based hydrogel degradation for the fabrication of vascular-derived microfluidic ...

Scaling of Engineered Vascular Grafts Using 3D Printed Guides and the Ring Stacking Method

工程血管移植缩放使用3D打印指南和环堆叠方法

Coronary artery disease remains a leading cause of death, affecting millions of Americans. With the lack of autologous vascular grafts available, ...