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

Эта работа финансируется Национальным институтом здоровья Грант R01 EB-008 836 и R01 HL083895 (как для LEN). Мы могли бы поблагодарить Дэрил Смит, стеклодув университета, для создания биореакторов для нашего исследования.

Автоклав Собрать и автоклава труб для системы потока и биореактор компонентов (биореактор себя и крышки силиконовой пробкой), как указано на рисунке 1 и рисунке 2. Кормление трубка имеет штекер на одном конце и открытый конец на другую сторону. Три коротких отрезков трубы в...

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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>

Качество инженерных судов в значительной степени диктуется качеством ГМК использоваться в культуре ткани. Важнейшие аспекты SMC фенотип включают сократительной морфологии, низкое число проход, и способность размножаться внутри биореактора. Мы рекомендуем пассаж быть не более P3 во вре...

<table border="1"><tbody><tr><th> Название реагента / Материал </th><th> Поставщик </th><th> Номер по каталогу </th></tr><tr><td> FBS (эмбриональной телячьей сыворотки) Тепло-инактивированная </td><td> Hyclone </td><td> SH30071 </td></tr><tr><td> DMEM </td><td> GIBCO </td><td> 11885 </td></tr><tr><td> rhFGF-основные </td><td> R &amp; B </td><td> 234-FSE </td></tr><tr><td> rrPDGF-BB </td><td> R &amp; B </td><td> 520-BB </td></tr><tr><td> Penicilin G </td><td> Сигма </td><td> Пенна </td></tr><tr><td> Медь (II) сульфат </td><td> Сигма </td><td> C8027 </td></tr><tr><td> Gylcine </td><td> Сигма </td><td> C8790 </td></tr><tr><td> L-аланин </td><td> Сигма </td><td> A7469-25G </td></tr><tr><td> L-пролин </td><td> Сигма </td><td> P5607-25G </td></tr><tr><td> Аскорбиновая кислота </td><td> Сигма </td><td> A4544-25G </td></tr><tr><td> HEPES </td><td> Сигма </td><td> H3375-100G </td></tr><tr><td> Силиконовая пробка </td><td> Коул-Пармер </td><td> 06298-24 </td></tr><tr><td> Masterflex труб L / S </td><td> Коул-Пармер </td><td> 06508-16, 06508-18 </td></tr><tr><td> Masterflex насоса </td><td> Коул-Пармер </td><td> 7553-80 </td></tr><tr><td> Dacron манжеты </td><td> Маке </td><td> 174406 </td></tr><tr><td> PGA чувствовал </td><td> Конкордия </td><td> MO000877-01 </td></tr><tr><td> 4-0 1,5 метрический Surgipro II шва </td><td> Syneture </td><td> VP-557-X </td></tr><tr><td> 6-0 0,7 метрический шва Дексон </td><td> Syneture </td><td> 7538-11 </td></tr><tr><td> 0.22μm PTFE фильтры </td><td> Ватман </td><td> 6780-2502 </td></tr><tr><td> Три Пути Стоп-кран </td><td> Эдвардс Lifesciences </td><td> 593WSC </td></tr><tr><td> Датчик давления </td><td> Эдвардс Lifesciences </td><td> PX212 </td></tr><tr><td> IV сумки </td><td> Бакстер </td><td> R4R2110 </td></tr><tr><td> Соленая набор разбавления </td><td> Стрелка </td><td> W20030 </td></tr><tr><td> Силиконовые трубки </td><td> Saint-Gobain </td><td> F05027 </td></tr></tbody></table>

engineering biological-based vascular grafts using a pulsatile bioreactor

Much effort has been devoted to develop and advance the methodology to regenerate functional small-diameter arterial bypasses. In the physiological environment, both mechanical and chemical stimulation are required to maintain the proper development and functionality of arterial vessels1,2.
Bioreactor culture systems developed by our group are designed to support vessel regeneration within a precisely controlled chemo-mechanical environment mimicking that of native vessels. Our bioreactor assembly and maintenance procedures are fairly simple and highly repeatable3,4. Smooth muscle cells (SMCs) are seeded onto a tubular polyglycolic acid (PGA) mesh that is threaded over compliant silicone tubing and cultured in the bioreactor with or without pulsatile stimulation for up to 12 weeks. There are four main attributes that distinguish our bioreactor from some predecessors. 1) Unlike other culture systems that simulate only the biochemical surrounding of native blood vessels, our bioreactor also creates a physiological pulsatile environment by applying cyclic radial strain to the vessels in culture. 2) Multiple engineered vessels can be cultured simultaneously under different mechanical conditions within a controlled chemical environment. 3) The bioreactor allows a mono layer of endothelial cells (EC) to be easily coated onto the luminal side of engineered vessels for animal implantation models. 4) Our bioreactor can also culture engineered vessels with different diameter size ranged from 1 mm to 3 mm, saving the effort to tailor each individual bioreactor to fit a specific diameter size. 
The engineered vessels cultured in our bioreactor resemble native blood vessels histologically to some degree. Cells in the vessel walls express mature SMC contractile markers such as smooth muscle myosin heavy chain (SMMHC)3. A substantial amount of collagen is deposited within the extracellular matrix, which is responsible for ultimate mechanical strength of the engineered vessels5. Biochemical analysis also indicates that collagen content of engineered vessels is comparable to that of native arteries6. Importantly, the pulsatile bioreactor has consistently regenerated vessels that exhibit mechanical properties that permit successful implantation experiments in animal models3,7. Additionally, this bioreactor can be further modified to allow real-time assessment and tracking of collagen remodeling over time, non-invasively, using a non-linear optical microscopy (NLOM)8. To conclude, this bioreactor should serve as an excellent platform to study the fundamental mechanisms that regulate the regeneration of functional small-diameter vascular grafts.

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

Наша группа разработала систему биореакторов культуры, который имитирует физиологическим стрессам пульсирующей сердечно-сосудистой системы к регенерации имплантируемых малого диаметра сосудистых трансплантатов.

Инженерно-биологической основе сосудистых трансплантатов использовании пульсирующего биореактора

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

Дизайн двухосных механических биореактор загрузка для тканевой инженерии

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

Operation of a Benchtop Bioreactor

Эксплуатация настольной биореактора

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

Приматов легких Decellularization и Recellularization помощью специализированного большого органа биореактор

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

Строительство и характеристика Fold биореактора Роман вокального

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

Выращивание клетках млекопитающих с использованием одноразовых Пневматический биореактора системы

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

Хирургическая техника для имплантации тканей Engineered сосудистые протезы и последующих<em&gt; В Vivo</em&gt; Мониторинг

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

Инженерные 3D Cellularized Коллаген Гели для сосудистой регенерации ткани

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

Роман биореактор для высокой плотности Выращивание разнородных микробных сообществ

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

Изображение наведением, на основе лазера Изготовление сосудисто-производных микрожидком сетей

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

Масштабирование Engineered сосудистых трансплантатов С помощью 3D печатных руководств и кольцо Метод Stacking

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