Name: micropatterning and assembly of 3d microvessels
Uploaded: 2016-09-09T15:00:00
Description: Watch this Scientific Journal Video about Micropatterning and Assembly of 3D Microvessels at JoVE.com

De auteurs willen graag de Lynn en Mike Garvey Imaging Laboratory aan het Instituut erkennen voor Stem Cell en Regenerative Medicine, evenals de Washington Nanofabrication Facility aan de Universiteit van Washington. Ze hebben ook de financiële steun van de National Institute of Health erkennen verleent DP2DK102258 (tot YZ) en training geeft T32EB001650 (tot SSK en MAR) en T32HL007312 (MAR).

1. Microfabricage van Patterned Polydimethylsiloxane (PDMS) met Network Design <ol><li> Wafer Fabrication een negatieve Template van het netwerk ontwerp Ontwerp <ol><li> Creëer een netwerk patroon met behulp van een computer-aided design (CAD) software. Controleer of de diagonale afmeting tussen de inlaat en uitlaat overeenkomen met de afstand tussen de inlaat en uitlaat reservoirs op de behuizing apparaten toekomstige stappen (zie 2.1.1). Opmerking: Het ontwerp van het patroon zelf wordt aang...

<ol>
	<li>Rubanyi, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+endothelium+in+cardiovascular+homeostasis+and+diseases.">The role of endothelium in cardiovascular homeostasis and diseases.</a> J. Cardiovasc. Pharmacol. 22, 37-44 (1993).</li><li>van Hinsbergh, V. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+endothelium:+vascular+control+of+haemostasis.">The endothelium: vascular control of haemostasis.</a> Eur. J. Obstet. Gynecol. Reprod. Biol. 95 (2), 198-201 (2001).</li><li>Chiu, J. -. J., Chien, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Disturbed+Flow+on+Vascular+Endothelium:+Pathophysiological+Basis+and+Clinical+Perspectives.">Effects of Disturbed Flow on Vascular Endothelium: Pathophysiological Basis and Clinical Perspectives.</a> Physiol. Rev. 91, 327-387 (2011).</li><li>Qi, Y., Jiang, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PDGF-BB+and+TGB-b1+on+cross-talk+between+endothelial+and+smooth+muscle+cells+in+vascular+remodeling+induced+by+low+shear+stress.">PDGF-BB and TGB-b1 on cross-talk between endothelial and smooth muscle cells in vascular remodeling induced by low shear stress.</a> Proc. Natl. Acad. Sci. U. S. A. 108, 1908-1913 (2011).</li><li>Sozzani, S., Del Prete, A., Bonecchi, R., Locati, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemokines+as+effector+and+target+molecules+in+vascular+biology.">Chemokines as effector and target molecules in vascular biology.</a> Cardiovasc. Res. 107 (3), 364-372 (2015).</li><li>Huh, D., Hamilton, G. A., Ingber, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+3D+cell+culture+to+organs-on-chips.">From 3D cell culture to organs-on-chips.</a> Trends Cell Biol. 21 (12), 745-754 (2011).</li><li>Staton, C. a., Reed, M. W. R., Brown, N. 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+critical+analysis+of+current+in+vitro+and+in+vivo+angiogenesis+assays.">A critical analysis of current in vitro and in vivo angiogenesis assays.</a> Int. J. Exp. Pathol. 90, 195-221 (2009).</li><li>Greek, R., Menache, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systematic+Reviews+of+Animal+Models:+Methodology+versus+Epistemology.">Systematic Reviews of Animal Models: Methodology versus Epistemology.</a> Int. J. Med. Sci. 10, 206-221 (2013).</li><li>van der Worp, H. B., Howells, D. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Can+Animal+Models+of+Disease+Reliably+Inform+Human+Studies.">Can Animal Models of Disease Reliably Inform Human Studies.</a> PLoS Med. 7 (3), e1000245 (2010).</li><li>Leong, X. -. F., Ng, C. -. Y., Jaarin, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+in+Cardiovascular+Research:+Hypertension+and+Atherosclerosis.">Animal Models in Cardiovascular Research: Hypertension and Atherosclerosis.</a> Biomed Res. Int. 2015, 528757 (2015).</li><li>Pries, A. R., Secomb, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Making+Microvascular+Networks+Work:+Angiogenesis,+Remodeling,+and+Pruning.">Making Microvascular Networks Work: Angiogenesis, Remodeling, and Pruning.</a> Physiology. 29, 446-455 (2014).</li><li>D'Amore, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+Of+Angiogenesis.">Mechanisms Of Angiogenesis.</a> Annu. Rev. Physiol. 49, 453-464 (1987).</li><li>Geudens, I., Gerhardt, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coordinating+cell+behaviour+during+blood+vessel+formation.">Coordinating cell behaviour during blood vessel formation.</a> Development. 138, 4569-4583 (2011).</li><li>Ribatti, D., Nico, B., Crivellato, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+of+the+vascular+system:+a+historical+overview.">The development of the vascular system: a historical overview.</a> Methods Mol. Biol. 1214, 1-14 (2015).</li><li>Ribatti, D., Nico, B., Crivellato, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphological+and+molecular+aspects+of+physiological+vascular+morphogenesis.">Morphological and molecular aspects of physiological vascular morphogenesis.</a> Angiogenesis. 12 (2), 101-111 (2009).</li><li>Bautch, V. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VEGF-directed+blood+vessel+patterning:+From+cells+to+organism.">VEGF-directed blood vessel patterning: From cells to organism.</a> Cold Spring Harb. Perspect. Med. 2 (9), 1-12 (2012).</li><li>Stratman, A. N., Schwindt, A. E., Malotte, K. M., Davis, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial-derived+PDGF-BB+and+HB-EGF+coordinately+regulate+pericyte+recruitment+during+vasculogenic+tube+assembly+and+stabilization.">Endothelial-derived PDGF-BB and HB-EGF coordinately regulate pericyte recruitment during vasculogenic tube assembly and stabilization.</a> Blood. 116, 4720-4730 (2010).</li><li>Bach, T. L., Barsigian, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VE-Cadherin+mediates+endothelial+cell+capillary+tube+formation+in+fibrin+and+collagen+gels.">VE-Cadherin mediates endothelial cell capillary tube formation in fibrin and collagen gels.</a> Exp. Cell Res. 238 (238), 324-334 (1998).</li><li>Kubow, K. E., Conrad, S. K., Horwitz, a. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrix+microarchitecture+and+myosin+II+determine+adhesion+in+3D+matrices.">Matrix microarchitecture and myosin II determine adhesion in 3D matrices.</a> Curr. Biol. 23 (17), 1607-1619 (2013).</li><li>Potapova, I. A., Gaudette, G. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchymal+Stem+Cells+Support+Migration,+Extracellular+Matrix+Invasion,+Proliferation,+and+Survival+of+Endothelial+Cells+In+Vitro.">Mesenchymal Stem Cells Support Migration, Extracellular Matrix Invasion, Proliferation, and Survival of Endothelial Cells In Vitro.</a> Stem Cells. 25 (7), 1761-1768 (2007).</li><li>Bayless, K. J., Davis, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sphingosine-1-phosphate+markedly+induces+matrix+metalloproteinase+and+integrin-dependent+human+endothelial+cell+invasion+and+lumen+formation+in+three-dimensional+collagen+and+fibrin+matrices.">Sphingosine-1-phosphate markedly induces matrix metalloproteinase and integrin-dependent human endothelial cell invasion and lumen formation in three-dimensional collagen and fibrin matrices.</a> Biochem. Biophys. Res. Commun. 312 (4), 903-913 (2003).</li><li>Hellstr&#246;m, M., Gerhardt, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lack+of+pericytes+leads+to+endothelial+hyperplasia+and+abnormal+vascular+morphogenesis.">Lack of pericytes leads to endothelial hyperplasia and abnormal vascular morphogenesis.</a> J. Cell Biol. 152 (3), 543-553 (2001).</li><li>Tulloch, N. L., Muskheli, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+of+Engineered+Human+Myocardium+With+Mechanical+Loading+and+Vascular+Coculture.">Growth of Engineered Human Myocardium With Mechanical Loading and Vascular Coculture.</a> Circ. Res. 109, 47-59 (2011).</li><li>Davis, G. E., Saunders, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+balance+of+capillary+tube+formation+versus+regression+in+wound+repair:+role+of+matrix+metalloproteinases+and+their+inhibitors.">Molecular balance of capillary tube formation versus regression in wound repair: role of matrix metalloproteinases and their inhibitors.</a> J. Investig. dermatology Symp. 11 (1), 44-56 (2006).</li><li>Kannan, R. Y., Salacinski, H. J., Butler, P. E., Hamilton, G., Seifalian, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+status+of+prosthetic+bypass+grafts:+a+review.">Current status of prosthetic bypass grafts: a review.</a> J. Biomed. Mater. Res. B. Appl. Biomater. 74, 570-581 (2005).</li><li>Nerem, R. M., Seliktar, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+Tissue+Engineering.">Vascular Tissue Engineering.</a> Annu. Rev. Biomed. Eng. 3 (1), 225-243 (2001).</li><li>Melchiorri, A. J., Hibino, N., Fisher, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strategies+and+techniques+to+enhance+the+in+situ+endothelialization+of+small-diameter+biodegradable+polymeric+vascular+grafts.">Strategies and techniques to enhance the in situ endothelialization of small-diameter biodegradable polymeric vascular grafts.</a> Tissue Eng. Part B. Rev. 19 (4), 292-307 (2013).</li><li>Abbott, W. M., Callow, A., Moore, W., Rutherford, R., Veith, F., Weinberg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+and+performance+standards+for+arterial+prostheses.">Evaluation and performance standards for arterial prostheses.</a> J. Vasc. Surg. 17 (4), 746-756 (1993).</li><li>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=Functional+Arteries+Grown+in+Vitro.">Functional Arteries Grown in Vitro.</a> Science. 284 (5413), 489-493 (1999).</li><li>Niklason, L., Counter, C. <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+-+Authors'+reply.">Blood vessels engineered from human cells - Authors' reply.</a> Lancet. 366 (9489), 892-893 (2005).</li><li>Zheng, Y., Chen, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+microvessels+for+the+study+of+angiogenesis+and+thrombosis.">In vitro microvessels for the study of angiogenesis and thrombosis.</a> Proc. Natl. Acad. Sci. U. S. A. 109, 9342-9347 (2012).</li><li>Zheng, Y., Chen, J., L&#242;pez, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow-driven+assembly+of+VWF+fibres+and+webs+in+in+vitro+microvessels.">Flow-driven assembly of VWF fibres and webs in in vitro microvessels.</a> Nat. Commun. 6, 7858 (2015).</li><li>Ligresti, G., Nagao, R. J., et al. <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+Three-Dimensional+Human+Peritubular+Microvascular+System.">A Novel Three-Dimensional Human Peritubular Microvascular System.</a> J. Am. Soc. Nephrol. 27, (2015).</li><li>Roberts, M. A., Tran, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stromal+cells+in+dense+collagen+promote+cardiomyocyte+and+microvascular+patterning+in+engineered+human+heart+tissue.">Stromal cells in dense collagen promote cardiomyocyte and microvascular patterning in engineered human heart tissue.</a> Tissue Eng. Part A. , (2016).</li><li>Qin, D., Xia, Y., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soft+lithography+for+micro-+and+nanoscale+patterning.">Soft lithography for micro- and nanoscale patterning.</a> Nat. Protoc. 5 (3), 491-502 (2010).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alpha-Step+200+Manual.">Alpha-Step 200 Manual.</a> Tencor Instruments. , (1989).</li><li>Rajan, N., Habermehl, J., Cot&#233;, M. -. F., Doillon, C. J., Mantovani, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+ready-to-use,+storable+and+reconstituted+type+I+collagen+from+rat+tail+tendon+for+tissue+engineering+applications.">Preparation of ready-to-use, storable and reconstituted type I collagen from rat tail tendon for tissue engineering applications.</a> Nat. Protoc. 1 (6), 2753-2758 (2006).</li><li>Baudin, B., Bruneel, A., Bosselut, N., Vaubourdolle, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+protocol+for+isolation+and+culture+of+human+umbilical+vein+endothelial+cells.">A protocol for isolation and culture of human umbilical vein endothelial cells.</a> Nat. Protoc. 2 (3), 481-485 (2007).</li><li>Leung, A. D., Wong, K. H. K., Tien, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasma+expanders+stabilize+human+microvessels+in+microfluidic+scaffolds.">Plasma expanders stabilize human microvessels in microfluidic scaffolds.</a> J. Biomed. Mater. Res. - Part A. 100 (7), 1815-1822 (2012).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tousimis+SAMDRI-780+Critical+Point+Drying+Apparatus.">Tousimis SAMDRI-780 Critical Point Drying Apparatus.</a> Tousimis Research Corporation. , (1987).</li><li>Palpant, N. J., Pabon, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+&#946;-catenin+signaling+respecifies+anterior-like+endothelium+into+beating+human+cardiomyocytes.">Inhibition of &#946;-catenin signaling respecifies anterior-like endothelium into beating human cardiomyocytes.</a> Development. 142 (18), 3198-3209 (2015).</li><li>Gimbrone, M. a., Topper, J. N., Nagel, T., Anderson, K. R., Garcia-Cardena, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+Dysfunction,+Hemodynamic+Forces,+and+Atherogenesis.">Endothelial Dysfunction, Hemodynamic Forces, and Atherogenesis.</a> Thromb. Haemost. 82, 722-726 (1999).</li><li>Wu, M. H., Ustinova, E., Granger, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrin+binding+to+fibronectin+and+vitronectin+maintains+the+barrier+function+of+isolated+porcine+coronary+venules.">Integrin binding to fibronectin and vitronectin maintains the barrier function of isolated porcine coronary venules.</a> J. Physiol. 532 (3), 785-791 (2001).</li><li>Ribatti, D., Nico, B., Vacca, A., Roncali, L., Dammacco, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+cell+heterogeneity+and+organ+specificity.">Endothelial cell heterogeneity and organ specificity.</a> J. Hematother. Stem Cell Res. 11, 81-90 (2002).</li><li>Shanks, N., Greek, R., Greek, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+animal+models+predictive+for+humans.">Are animal models predictive for humans.</a> Philos. Ethics. Humanit. Med. 4, 2 (2009).</li></ol>

Gemanipuleerde microvaatjes een in vitro model waarbij fysiologische karakteristieken zoals luminale geometrie, hydrodynamische krachten en meercellige interacties aanwezig zijn en instelbaar zijn. Dit type platform is krachtig omdat het de mogelijkheid biedt te modelleren en bestuderen endotheliale gedrag in verschillende contexten, waar de in vitro kweekomstandigheden kunnen worden afgestemd op dat van de micro betrokken. Bijvoorbeeld, de mechanismen die endotheliale processen, zoals angiogenese, bek...

De microvasculatuur in elk orgaan helpt bepalen het weefsel micro, onderhouden weefsel homeostase en reguleren ontsteking, permeabiliteit, trombose, en fibrinolyse 1,2. Microvasculaire endotheel, in het bijzonder is de interface tussen de bloedstroom en het omliggende weefsel en daarom een cruciale rol bij het ​​moduleren van vasculaire en orgaanfunctie reactie op stimuli zoals hydrodynamische krachten en circulerende cytokinen en hormonen speelt 3 - 5. Het begrijpen van de gedetailleerde interacties tussen het endotheel, bloed en weefsel micro belangrijk voor de studie van vasculaire biologie en ziekteprogressie. Echter, is vooruitgang geboekt bij het ​​bestuderen van deze interacties werd gehinderd door een beperkte in vitro hulpmiddelen die niet herhalen in vivo microvasculaire structuur en functie 6,7. Daardoor heeft het veld en therapeutische vooruitgang sterk aangevoerde kostbare en tijd-consumeren diermodellen die vaak niet te vertalen naar succes bij mensen 8-10. Terwijl in vivo modellen zijn van onschatbare waarde bij het ​​bestuderen van ziektemechanismen en vasculaire functies, ze complex en vaak geen nauwkeurige controle van individuele cellen, biochemische en biofysische signalen. Bloedvaten door het hele lichaam beschikt over een volwassen hiërarchische structuur in combinatie met uitgestrekte capillaire bedden, het verstrekken van optimale doorbloeding en voedingsstoffen transport gelijktijdig 11. Aanvankelijk vasculatuur vormen een primitieve plexus die reorganiseert een hiërarchisch vertakte netwerk tijdens vroege ontwikkeling 12,13. Hoewel veel van de bij deze processen signalen goed begrepen 14-16, blijft ongrijpbaar hoe dergelijke vasculaire patroonvorming 15 bepaald. Op zijn beurt, recapituleren dit proces in vitro georganiseerde vasculaire netwerken ingenieur heeft been moeilijke Veel bestaande in vitro platforms. model vaatstelsel, zoals tweedimensionale endotheliale celculturen missen belangrijke kenmerken zoals meercellige nabijheid luminale driedimensionale geometrie, stroom en extracellulaire matrix. Buisvorming assays 3D hydrogels (collageen of fibrine) 17-19 of invasie assays 20,21 zijn gebruikt om de endotheelfunctie bij 3D en hun interacties met andere vasculaire bestuderen 17,22 of weefsel celtypes 23. Echter, geassembleerd lumens in deze testen missen interconnectiviteit, hemodynamische stroom en geschikte perfusie. Bovendien is de neiging tot vasculaire regressie deze buisvorming assays 24 voorkomt lange termijn cultuur en rijping die de mate van functionele studies die uitgevoerd kunnen worden beperkt. Er is dus een groeiende behoefte om in vitro platforms van microvasculaire netwerken die goed kunnen modelleren en engineeringdothelial eigenschappen en kunnen langdurige kweek. Verschillende vasculaire manipulatietechnieken zijn opgekomen der jaren voor medische toepassingen te vervangen of bypass getroffen vaten bij patiënten met vasculaire ziekte. Grote diameter vaartuigen gemaakt van synthetische materialen zoals polyethyleentereftalaat (PET), en polytetrafluorethyleen (ePTFE) hebben aanzienlijke therapeutische succes op lange termijn patency (gemiddeld 95% patency dan 5 jaar) 25 had. Hoewel klein diameter synthetische transplantaten (&lt;6 mm), meestal te kampen hebben complicaties, zoals intima hyperplasie en trombopoïese 26-28, tissue engineered grafts kleine diameter gemaakt met biologisch materiaal hebben aanzienlijke vooruitgang 29,30 gemaakt. Ondanks de vooruitgang van dit soort, zijn gemanipuleerd schepen op de microschaal een uitdaging gebleven. Adequaat model van de microvasculatuur, is het noodzakelijk om complexe netwerkpatronen met suf genererendoende mechanische sterkte doorgankelijkheid en een matrixsamenstelling die het mogelijk maakt voor zowel nutriënten permeatie voor parenchymale cellen en cellulaire remodeling handhaven. Dit protocol bevat een nieuwe kunstmatige perfusable vaartuig netwerk dat een inwoner in vivo instelling met een instelbare en controleerbare micro 31 nabootst - 34. De beschreven werkwijze genereert Engineered microvaatjes met een diameter in de orde van 100 urn. Engineered microvaten worden vervaardigd door perfuseren endotheelcellen door middel van een microfluïdische kanaal, dat is ingebed in zacht type I collageen hydrogel. Dit systeem heeft het vermogen om een ​​patroon netten met open luminale structuur genereren repliceren meercellige interacties moduleren extracellulaire matrix samenstelling en fysiologisch relevante hemodynamische krachten toegepast.

<table border="0" cellpadding="0" cellspacing="0" width="1363">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:459px;">Wafer Fabrication</td>
			<td style="width:225px;"></td>
			<td style="width:140px;"></td>
			<td style="width:540px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">AutoGlow Plasma System</td>
			<td>AutoGlow</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Headway Spin Coater</td>
			<td>Headway Research, Inc&#160;</td>
			<td colspan="2">PWM32 Spin Coater&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">ABM Contact Aligner</td>
			<td>AB-M</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Alpha Step Profilometer</td>
			<td>Tencor</td>
			<td>Alpha Step 200</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SU-8 Developer</td>
			<td>Microchem</td>
			<td>Y020100</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SU-8 Resist</td>
			<td>Microchem</td>
			<td>SU-8 2000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">8&#34; silicon wafer</td>
			<td>Wafer World Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tabletop Micro Pattern Generator</td>
			<td>Heidelberg Instruments</td>
			<td>&#956;PG 101</td>
			<td>For generation of photomask</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Hot plate</td>
			<td>VWR</td>
			<td>97042-646</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ispropyl alcohol</td>
			<td>Avantor Performance Materials</td>
			<td>9088</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Petri dishes (120 x 120 mm, square)</td>
			<td>Sigma-Aldrich</td>
			<td>Z617679</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Trichloro(3,3,3-trifluoropropyl)silane</td>
			<td>Sigma-Aldrich</td>
			<td>MKBG3805V</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Polydimethylsiloxane (PDMS) elastomer base and curing agent</td>
			<td>Dow Corning</td>
			<td>Sylgard 184</td>
			<td>Mixed at 10:1 (w/w)</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Vacuum desiccator</td>
			<td>Sigma-Aldrich</td>
			<td>Z119024-1EA</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Oven</td>
			<td>VWR</td>
			<td>9120976</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Device Fabrication and Culture</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">poly(methyl methacrylate) (PMMA)</td>
			<td>Plexiglas</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Corona Treater</td>
			<td>Electro-Technic Products, Inc.</td>
			<td>BD-20</td>
			<td>Handheld device for plasma treatment of PMMA devices and PDMS molds</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Soldering Iron</td>
			<td>Weller&#160;</td>
			<td>WTCPS</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Stainless Steel Truss Head Slotted Machine Screw</td>
			<td>McMaster-Carr&#160;</td>
			<td>91785A096</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Stainless steel dowel pins</td>
			<td>McMaster-Carr&#160;</td>
			<td>93600A060</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tweezers&#160;</td>
			<td>Miltex</td>
			<td>24-572</td>
			<td>Any similar tweezers may be used</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Spatula (Micro Spoon)</td>
			<td>Electron Microscopy Services</td>
			<td>62410-01</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Screw driver</td>
			<td></td>
			<td></td>
			<td>Any flat head screwdriver may be used, autoclaved</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Glass coverslips (22 x 22 mm)</td>
			<td>Fisher Scientific</td>
			<td>12-542B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bleach</td>
			<td>Clorox</td>
			<td>4460030966</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Petri dishes (150 X 25mm)</td>
			<td>Corning</td>
			<td>430599</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Petri dishes (100 X 20 mm)</td>
			<td>Corning</td>
			<td>2909</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cotton, cut into 1 cm x 3 cm pieces</td>
			<td></td>
			<td></td>
			<td>Autoclaved</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Polyethyleneimine (PEI)</td>
			<td>Sigma-Aldrich</td>
			<td>P3143</td>
			<td>Dilute to 1% in cell culture grade water</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Glutaraldehyde</td>
			<td>Sigma-Aldrich</td>
			<td>G6257</td>
			<td>Dilute to 0.1% in cell culture grade water</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">Sterile H2O</td>
			<td></td>
			<td></td>
			<td>Autoclaved DI H2O</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Type I collagen, dissolved in 0.1% acetic acid</td>
			<td></td>
			<td></td>
			<td>Isolated from rat tails as described in Rajan et. al. 2006 (ref #37)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">1 mL syringe</td>
			<td>BD</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">10 mL syringe</td>
			<td>BD</td>
			<td>309604</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">15 mL conical tubes</td>
			<td>Corning</td>
			<td>352097</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">30 mL conical tubes</td>
			<td>Corning</td>
			<td>352098</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">M199 10X Media&#160;</td>
			<td>Life Technologies</td>
			<td>11825-015</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">1N NaOH (sterile)</td>
			<td>Sigma-Aldrich</td>
			<td>415413</td>
			<td>Dilute to 1N in cell culture grade water</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">HUVECs&#160;</td>
			<td>Lonza</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Endothelial growth media</td>
			<td>Lonza</td>
			<td>CC-3124</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Trypsin</td>
			<td>Corning</td>
			<td>25-052-CI</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fetal bovine serum (FBS)</td>
			<td>Thermofisher Scientific</td>
			<td>10082147</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dextran from Leuconostoc spp. (70kDa)</td>
			<td>Sigma-Aldrich</td>
			<td>31390</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Phosphate Buffered Saline (PBS)</td>
			<td>Corning</td>
			<td>21-031-CV</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Hemocytometer</td>
			<td>Hausser Scientific Co.</td>
			<td>3200</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Gel loading tips</td>
			<td>VWR</td>
			<td>37001-152</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">18G Blunt Fill Needle</td>
			<td>BD&#160;</td>
			<td>305180</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">20G Stainless Steel Dispensing Needle</td>
			<td>McMaster-Carr</td>
			<td>75165A123</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tygon 1/32&#8221; ID, 3/32&#34; OD Silicon Tubing</td>
			<td>Cole-Parmer</td>
			<td>EW-95702-00</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">1/16&#34; Tube-to-tube Coupling</td>
			<td>McMaster-Carr</td>
			<td>5116K165</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">90&#176; Elbow Connectors, Tube-to-Tube</td>
			<td>McMaster-Carr</td>
			<td>5121K901</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Luer Lock Coupling (Female, 1/16&#34; ID)</td>
			<td>McMaster-Carr</td>
			<td>51525K211</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Plastic Forceps, with Jaw Grips</td>
			<td>Electron Microscopy Services</td>
			<td>72971</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dual Syringe Pump</td>
			<td>Harvard Apparatus</td>
			<td>70-4505</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">5 mL Polystyrene Round-bottom tube</td>
			<td>Fisher Scientific</td>
			<td>14-959-2A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Device Analysis</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Formaldehyde</td>
			<td>Sigma-Aldrich</td>
			<td>F8775</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bovine Serum Albumin</td>
			<td>Sigma-Aldrich</td>
			<td>A8806-5G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Triton X-100</td>
			<td>Sigma-Aldrich</td>
			<td>T-9284</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Rabbit anti-hCD31</td>
			<td>Abcam</td>
			<td>ab32457</td>
			<td>1:25 working dilution</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">FITC conjugated anti-von Willebrand Factor antibody</td>
			<td>Abcam</td>
			<td>ab8822</td>
			<td>1:100 working dilution</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Goat anti-rabbit 568 secondary antibody</td>
			<td>Thermofisher Scientific</td>
			<td>A-11011</td>
			<td>1:100 working dilution</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Hoescht</td>
			<td>Thermofisher Scientific</td>
			<td>H1399</td>
			<td>Resuspended in DMSO</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Sodium cacodylate&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>C0250</td>
			<td>To make 0.2M cacodylate buffer</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ethanol</td>
			<td>VWR International</td>
			<td>BDH1164-4LP</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">40kDa FITC-conjugated Dextran</td>
			<td>Sigma-Aldrich</td>
			<td>FD40S&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Additional Culture Reagents&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">CHIR-99021</td>
			<td>Selleck Chem</td>
			<td>S2924</td>
			<td>Small molecule GSK-3 inhibitor</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Human recombinant VEGF</td>
			<td>Peprotech</td>
			<td>100-20</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Human recombinant bFGF</td>
			<td>Peprotech</td>
			<td>AF-100-18B</td>
			<td></td>
		</tr>
	</tbody>
</table>

micropatterning and assembly of 3d microvessels

In vitro studie platforms endotheelcellen en vasculaire biologie grotendeels beperkt tot 2D endotheliale celculturen, stroomkamers met polymeer of glazen ondergronden en hydrogel gebaseerde vaatvorming assays. Deze testen, terwijl informatief, niet herhalen lumen geometrie, de juiste extracellulaire matrix, en meercellige nabijheid, die een belangrijke rol bij het moduleren vaatfunctie spelen. Dit manuscript beschrijft een spuitgietwerkwijze met technisch vaartuigen met een diameter in de orde van 100 urn genereren. Microvaten worden vervaardigd door het zaaien van endotheelcellen in een microfluïdische kanaal ingebed in een native type I collageen hydrogel. Door de integratie van parenchymcellen in het collageen matrix voorafgaand aan de vorming van het kanaal, kan specifiek weefsel micro-omgevingen worden gemodelleerd en bestudeerd. Extra modulaties van hydrodynamische eigenschappen en media samenstelling zorgen voor controle van complexe vasculaire functie binnen de gewenste micro-omgeving.Dit platform maakt de studie van perivasculaire celrecrutering, bloed-endotheel interacties stroomreactie en weefselspecifieke microvasculaire interacties. Engineered microvaatjes bieden de mogelijkheid om de invloed van de individuele componenten van een vasculaire niche isoleren en nauwkeurig te regelen zijn chemische, mechanische en biologische eigenschappen vasculaire biologie bestuderen in zowel gezondheid en ziekte.

De gemanipuleerde vat platform creëert functionele microvasculatuur ingebed in een natuurlijke collageen type I matrix en zorgt voor strakke controle van de cellulaire, biochemische en biofysische milieu in vitro. Tot complete microvaten fabriceren, worden humane navelstreng endotheelcellen (HUVEC) geperfuseerd via het collageen geïntegreerde microfluïdische netwerk waar zij hechten aan octrooi lumen en confluent endotheel vormen. Zoals geïllustreerd in figuur 1A-C,<...

Watch this Scientific Journal Video about Micropatterning and Assembly of 3D Microvessels at JoVE.com

Micropatterning and Assembly of 3D Microvessels

Dit manuscript bevat een spuitgiet methode om microvaten dat fysiologische eigenschappen van endotheel recapituleren ingenieur. De microfluïdische-based proces creëert patent 3D vasculaire netwerken met tailorable omstandigheden, zoals stroom, cellulaire samenstelling, geometrie, en biochemische hellingen. Het fabricageproces en voorbeelden van mogelijke toepassingen worden beschreven.

Micropatterning en montage van 3D microvaten

In vitro platforms to study endothelial cells and vascular biology are largely limited to 2D endothelial cell culture, flow chambers with polymer or glass ...

Research

JoVE Journal

Bioengineering

Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo

Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo

Voorbereiding van de 3D Collageen Gels en Microkanalen voor de Studie van 3D-Interacties<em&gt; In Vivo</em

Historically, most cellular processes have been studied in only 2 dimensions.  While these studies have been informative about general cell signaling ...

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Geautomatiseerde robot Liquid Handling samenstel van modulaire DNA voorzieningen

Recent advances in modular DNA assembly techniques have enabled synthetic biologists to test significantly more of the available &#34;design space&#34; ...

Synthesis of Cationized Magnetoferritin for Ultra-fast Magnetization of Cells

Synthese van gekationiseerde Magnetoferritin voor ultrasnelle magnetisatie van Cells

Many important biomedical applications, such as cell imaging and remote manipulation, can be achieved by labeling cells with superparamagnetic iron oxide ...

Bioprinting of Cartilage and Skin Tissue Analogs Utilizing a Novel Passive Mixing Unit Technique for Bioink Precellularization

Bioprinting van kraakbeen en huid weefsel analogen met behulp van een roman passief mengen eenheid techniek voor Bioink Precellularization

Bioprinting is a powerful technique for the rapid and reproducible fabrication of constructs for tissue engineering applications. In this study, both ...

Fabrication of Custom Agarose Wells for Cell Seeding and Tissue Ring Self-assembly Using 3D-Printed Molds

Fabricage van aangepaste Agarose putten voor het zaaien van de cellen en weefsel Ring zelf-assemblage met behulp van 3D-gedrukte mallen

Engineered tissues are being used clinically for tissue repair and replacement, and are being developed as tools for drug screening and human disease ...

Protocols of 3D Bioprinting of Gelatin Methacryloyl Hydrogel Based Bioinks

Protocollen van 3D Bioprinting van gelatine Methacryloyl hydrogel gebaseerde Bioinks

Gelatin methacryloyl (GelMA) has become a popular biomaterial in the field of bioprinting. The derivation of this material is gelatin, which is hydrolyzed ...

Temperature-Controlled Assembly and Characterization of a Droplet Interface Bilayer

Temperatuurgecontroleerde montage en karakterisering van een druppelinterface-bilayer

The droplet interface bilayer (DIB) method for assembling lipid bilayers (i.e., DIBs) between lipid-coated aqueous droplets in oil offers key benefits ...

Control of Cell Geometry through Infrared Laser Assisted Micropatterning

Controle van celgeometrie door infrarood laserondersteunde micropatterning

Micropatterning is an established technique in the cell biology community used to study connections between the morphology and function of cellular ...

A Micropatterning Assay for Measuring Cell Chirality

Een micropatterning-test voor het meten van celchiraliteit

Chirality is an intrinsic cellular property, which depicts the asymmetry in terms of polarization along the left-right axis of the cell. As this unique ...

Gelatin Methacryloyl Granular Hydrogel Scaffolds: High-throughput Microgel Fabrication, Lyophilization, Chemical Assembly, and 3D Bioprinting

Gelatine Methacryloyl Granular Hydrogel Scaffolds: High-throughput Microgel Fabrication, Lyophilization, Chemical Assembly, en 3D Bioprinting

The emergence of granular hydrogel scaffolds (GHS), fabricated via assembling hydrogel microparticles (HMPs), has enabled microporous scaffold formation ...