Name: bioprintable alginate/gelatin hydrogel 3d in vitro model systems induce cell spheroid formation
Uploaded: 2018-07-02T14:30:00
Description: Watch this Scientific Journal Video about Bioprintable Alginate/Gelatin Hydrogel 3D In Vitro Model Systems Induce Cell Spheroid Formation at JoVE.com

Tao Jiang Çin burs Konseyi (201403170354) ve McGill Mühendisliği doktora Ödülü (90025) kendi burs finansman için teşekkürler. Jose G. Munguia-Lopez CONACYT (250279, 290936 ve 291168) ve FRQNT (258421) kendi burs finansman için teşekkürler. Salvador Flores-Torres CONACYT (751540) finansman onların burs için teşekkürler. Joseph M. Kinsella Ulusal Bilim ve Mühendislik Araştırma Konseyi, Kanada Vakfı yenilik, Townshend-Lamarre Aile Vakfı ve McGill Üniversitesi için kendi finansman için teşekkürler. Allen Ehrlicher onun rheometer Dan bize fluorescently etiketli hücre hatları için erişim vermek için onun confocal mikroskop ve Morag Park kullanmak bize izin verdiği Nicolau kullanmak için bize izin verdiği için teşekkür etmek istiyorum.

1. malzeme, hidrojel ve hücre kültür malzemelerin hazırlanması
<ol><li>
Malzeme ve eriyik hazırlığı
	<ol>
<li>Yıkayıp kurulayın 250 mL ve 100 mL Cam şişeler, manyetik karıştırıcı, spatula, 10 mL kartuş, 25 G silindirik püskürtme (0.5 uzunluğunda) ve 250 µm bir iç çapı ile. Malzemeler tarafından ısıyla sterilize 121 ° C/15 dk onları/1 atm. tutmak malzemeler steril koşullarda kadar kullanmak. Not: satıcı bilgileri Tablo reçetesi bakın.</li>
		<li>A...

<ol>
	<li>Cui, X., Hartanto, Y., Zhang, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+multicellular+spheroids+formation.">Advances in multicellular spheroids formation.</a> Journal of the Royal Society Interface. 14 (127), (2017).</li><li>Yip, D., Cho, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+multicellular+3D+heterospheroid+model+of+liver+tumor+and+stromal+cells+in+collagen+gel+for+anti-cancer+drug+testing.">A multicellular 3D heterospheroid model of liver tumor and stromal cells in collagen gel for anti-cancer drug testing.</a> Biochemical and Biophysical Research Communications. 433 (3), 327-332 (2013).</li><li>Breslin, S., O'Driscoll, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relevance+of+using+3D+cell+cultures,+in+addition+to+2D+monolayer+cultures,+when+evaluating+breast+cancer+drug+sensitivity+and+resistance.">The relevance of using 3D cell cultures, in addition to 2D monolayer cultures, when evaluating breast cancer drug sensitivity and resistance.</a> Oncotarget. 7 (29), 45745-45756 (2016).</li><li>Yue, X., Lukowski, J. K., Weaver, E. M., Skube, S. B., Hummon, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+proteomic+and+phosphoproteomic+comparison+of+2D+and+3D+colon+cancer+cell+culture+models.">Quantitative proteomic and phosphoproteomic comparison of 2D and 3D colon cancer cell culture models.</a> Journal of Proteome Research. 15 (12), 4265-4276 (2016).</li><li>Priwitaningrum, D. 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=Tumor+stroma-containing+3D+spheroid+arrays:+a+tool+to+study+nanoparticle+penetration.">Tumor stroma-containing 3D spheroid arrays: a tool to study nanoparticle penetration.</a> Journal of Controlled Release. 244 (Pt B), 257-268 (2016).</li><li>Hong, S., 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=Cellular+behavior+in+micropatterned+hydrogels+by+bioprinting+system+depended+on+the+cell+types+and+cellular+interaction.">Cellular behavior in micropatterned hydrogels by bioprinting system depended on the cell types and cellular interaction.</a> Journal of Bioscience and Bioengineering. 116 (2), 224-230 (2013).</li><li>Dolati, F., 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+evaluation+of+carbon-nanotube-reinforced+bioprintable+vascular+conduits.">In vitro evaluation of carbon-nanotube-reinforced bioprintable vascular conduits.</a> Nanotechnology. 25 (14), 145101 (2014).</li><li>Murphy, S. V., Atala, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D+bioprinting+of+tissues+and+organs.">3D bioprinting of tissues and organs.</a> Nature Biotechnology. 32 (8), 773-785 (2014).</li><li>Kang, H. 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=A+3D+bioprinting+system+to+produce+human-scale+tissue+constructs+with+structural+integrity.">A 3D bioprinting system to produce human-scale tissue constructs with structural integrity.</a> Nature Biotechnology. 34 (3), 312-319 (2016).</li><li>Miller, J. S., 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=Rapid+casting+of+patterned+vascular+networks+for+perfusable+engineered+three-dimensional+tissues.">Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues.</a> Nature Materials. 11 (9), 768-774 (2012).</li><li>Jia, 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=Direct+3D+bioprinting+of+perfusable+vascular+constructs+using+a+blend+bioink.">Direct 3D bioprinting of perfusable vascular constructs using a blend bioink.</a> Biomaterials. 106, 58-68 (2016).</li><li>Kolesky, D. B., Homan, K. A., Skylar-Scott, M. A., Lewis, 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=Three-dimensional+bioprinting+of+thick+vascularized+tissues.">Three-dimensional bioprinting of thick vascularized tissues.</a> Proceedings of the National Academy of Sciences of the United States of America. 113 (12), 3179-3184 (2016).</li><li>Lee, 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=Design+and+fabrication+of+human+skin+by+three-dimensional+bioprinting.">Design and fabrication of human skin by three-dimensional bioprinting.</a> Tissue Engineering Part C: Methods. 20 (6), 473-484 (2014).</li><li>Jiang, T., 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=Directing+the+self-assembly+of+tumour+spheroids+by+bioprinting+cellular+heterogeneous+models+within+alginate/gelatin+hydrogels.">Directing the self-assembly of tumour spheroids by bioprinting cellular heterogeneous models within alginate/gelatin hydrogels.</a> Scientific Reports. 7 (1), 4575 (2017).</li><li>Knowlton, S., Onal, S., Yu, C. H., Zhao, J. J., Tasoglu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioprinting+for+cancer+research.">Bioprinting for cancer research.</a> Trends in Biotechnology. 33 (9), 504-513 (2015).</li><li>Derby, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Printing+and+prototyping+of+tissues+and+scaffolds.">Printing and prototyping of tissues and scaffolds.</a> Science. 338 (6109), 921-926 (2012).</li><li>Nair, K., 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=Characterization+of+cell+viability+during+bioprinting+processes.">Characterization of cell viability during bioprinting processes.</a> Biotechnology Journal. 4 (8), 1168-1177 (2009).</li><li>Costa, E. 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=3D+tumor+spheroids:+an+overview+on+the+tools+and+techniques+used+for+their+analysis.">3D tumor spheroids: an overview on the tools and techniques used for their analysis.</a> Biotechnology Advances. 34 (8), 1427-1441 (2016).</li><li>Zhao, Y., 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=Three-dimensional+printing+of+Hela+cells+for+cervical+tumor+model+in+vitro.">Three-dimensional printing of Hela cells for cervical tumor model in vitro.</a> Biofabrication. 6 (3), 035001 (2014).</li><li>Ling, K., 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=Bioprinting-based+high-throughput+fabrication+of+three-dimensional+MCF-7+human+breast+cancer+cellular+spheroids.">Bioprinting-based high-throughput fabrication of three-dimensional MCF-7 human breast cancer cellular spheroids.</a> Engineering. 1 (2), 269-274 (2015).</li><li>Liang, Y., 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+cell-instructive+hydrogel+to+regulate+malignancy+of+3D+tumor+spheroids+with+matrix+rigidity.">A cell-instructive hydrogel to regulate malignancy of 3D tumor spheroids with matrix rigidity.</a> Biomaterials. 32 (35), 9308-9315 (2011).</li><li>Szot, C. S., Buchanan, C. F., Freeman, J. W., Rylander, M. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D+in+vitro+bioengineered+tumors+based+on+collagen+I+hydrogels.">3D in vitro bioengineered tumors based on collagen I hydrogels.</a> Biomaterials. 32 (31), 7905-7912 (2011).</li><li>Carey, S. P., Kraning-Rush, C. M., Williams, R. M., Reinhart-King, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biophysical+control+of+invasive+tumor+cell+behavior+by+extracellular+matrix+microarchitecture.">Biophysical control of invasive tumor cell behavior by extracellular matrix microarchitecture.</a> Biomaterials. 33 (16), 4157-4165 (2012).</li><li>Hospodiuk, M., Dey, M., Sosnoski, D., Ozbolat, I. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+bioink:+a+comprehensive+review+on+bioprintable+materials.">The bioink: a comprehensive review on bioprintable materials.</a> Biotechnology Advances. 35 (2), 217-239 (2017).</li><li>Caliari, S. R., Burdick, 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=A+practical+guide+to+hydrogels+for+cell+culture.">A practical guide to hydrogels for cell culture.</a> Nature Methods. 13 (5), 405-414 (2016).</li><li>Bhutani, U., Laha, A., Mitra, K., Majumdar, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sodium+alginate+and+gelatin+hydrogels:+viscosity+effect+on+hydrophobic+drug+release.">Sodium alginate and gelatin hydrogels: viscosity effect on hydrophobic drug release.</a> Materials Letters. 164, 76-79 (2016).</li><li>Biswal, 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=Effect+of+mechanical+and+electrical+behavior+of+gelatin+hydrogels+on+drug+release+and+cell+proliferation.">Effect of mechanical and electrical behavior of gelatin hydrogels on drug release and cell proliferation.</a> Journal of the Mechanical Behavior of Biomedical Materials. 53, 174-186 (2016).</li><li>Rowley, J. A., Madlambayan, G., Mooney, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alginate+hydrogels+as+synthetic+extracellular+matrix+materials.">Alginate hydrogels as synthetic extracellular matrix materials.</a> Biomaterials. 20 (1), 45-53 (1999).</li><li>Djabourov, M., Leblond, J., Papon, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gelation+of+aqueous+gelatin+solutions.+I.+Structural+investigation.">Gelation of aqueous gelatin solutions. I. Structural investigation.</a> Journal de Physique (France). 49 (2), 319-332 (1988).</li><li>Djabourov, M., Leblond, J., Papon, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gelation+of+aqueous+gelatin+solutions.+II.+Rheology+of+the+sol-gel+transition.">Gelation of aqueous gelatin solutions. II. Rheology of the sol-gel transition.</a> Journal de Physique (France). 49 (2), 333-343 (1988).</li><li>Coussot, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Rheometry of Pastes, Suspensions, and Granular Materials: Applications in Industry and Environment. , (2005).</li><li>Ouyang, L., Yao, R., Zhao, Y., Sun, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+bioink+properties+on+printability+and+cell+viability+for+3D+bioplotting+of+embryonic+stem+cells.">Effect of bioink properties on printability and cell viability for 3D bioplotting of embryonic stem cells.</a> Biofabrication. 8 (3), 035020 (2016).</li><li>Michon, C., Cuvelier, G., Launay, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Concentration+dependence+of+the+critical+viscoelastic+properties+of+gelatin+at+the+gel+point.">Concentration dependence of the critical viscoelastic properties of gelatin at the gel point.</a> Rheologica Acta Rheologica Acta: An International Journal of Rheology. 32 (1), 94-103 (1993).</li><li>Mouser, V. 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=Yield+stress+determines+bioprintability+of+hydrogels+based+on+gelatin-methacryloyl+and+gellan+gum+for+cartilage+bioprinting.">Yield stress determines bioprintability of hydrogels based on gelatin-methacryloyl and gellan gum for cartilage bioprinting.</a> Biofabrication. 8 (3), 035003 (2016).</li><li>Benbow, J. J., Oxley, E. W., Bridgwater, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+extrusion+mechanics+of+pastes-the+influence+of+paste+formulation+on+extrusion+parameters.">The extrusion mechanics of pastes-the influence of paste formulation on extrusion parameters.</a> Chemical Engineering Science. 42 (9), 2151-2162 (1987).</li><li>Bingham, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Fluidity and plasticity. , (1922).</li><li>Horrobin, D. J., Nedderman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Die+entry+pressure+drops+in+paste+extrusion.">Die entry pressure drops in paste extrusion.</a> Chemical Engineering Science. 53 (18), 3215-3225 (1998).</li><li>Soman, P., 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=Cancer+cell+migration+within+3D+layer-by-layer+microfabricated+photocrosslinked+PEG+scaffolds+with+tunable+stiffness.">Cancer cell migration within 3D layer-by-layer microfabricated photocrosslinked PEG scaffolds with tunable stiffness.</a> Biomaterials. 33 (29), 7064-7070 (2012).</li><li>Asghar, 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=Engineering+cancer+microenvironments+for+in+vitro+3-D+tumor+models.">Engineering cancer microenvironments for in vitro 3-D tumor models.</a> Materials Today. 18 (10), 539-553 (2015).</li><li>Lin, R. Z., Chang, H. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+three-dimensional+multicellular+spheroid+culture+for+biomedical+research.">Recent advances in three-dimensional multicellular spheroid culture for biomedical research.</a> Biotechnology Journal. 3 (9-10), 1172-1184 (2008).</li><li>Akasov, 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=Formation+of+multicellular+tumor+spheroids+induced+by+cyclic+RGD-peptides+and+use+for+anticancer+drug+testing+in+vitro.">Formation of multicellular tumor spheroids induced by cyclic RGD-peptides and use for anticancer drug testing in vitro.</a> International Journal of Pharmaceutics. 506 (1-2), 148-157 (2016).</li></ol>

Kirlenme (biyolojik ya da kimyasal) sürecinde herhangi bir noktada oluşursa hücre yüklü yapıları tehlikeye girebilir. Genellikle, biyolojik kirlenme iki sonra görülür veya kültür medya veya bioprinted yapı ile üç gün kültür bir renk olarak değiştirin. Bu nedenle, sterilizasyon (fiziksel ve kimyasal dezenfeksiyon) tüm hücre ile ilgili işlemler için önemli bir adımdır. Önemli, ısıyla jelatin daha yavaş jel biz yürütülen çalışmalarda yapılan jelleşme özelliklerini değiştirir. Bu ned...

2D hücre kültürü yaygın kanser araştırmalarında kullanılan rağmen hücrelere besin ve oksijen Tekdüzen bir konsantrasyon monolayer biçimiyle yetiştirilen gibi sınırlamalar var. Bu kültürler eksikliği önemli hücre-hücre ve hücre-matris etkileşimleri yerli tümör microenvironment (TME) mevcut. Sonuç olarak, bu modeller özetlemek kötü sonuçlanan anormal hücre davranışları doğal olmayan türleri morfoloji, düzensiz reseptör organizasyon, membran polarizasyon ve aralarında anormal gen ekspresyonu da dahil olmak üzere, fizyolojik şartlarda koşulları1,2,3,4. Öte yandan, 3D hücre kültürü, nerede hücreleri hacimsel bir alanda toplamları, pulcuklarının veya organoids genişletilir, temel hücre biyolojisi ve Fizyoloji çalışmaya daha doğru vitro ortamlar oluşturmak için alternatif bir teknik sunar. 3D hücre kültür modelleri aynı zamanda yerel TME vitro1,4,5kritik fizyolojik özellikleri hücre-ECM etkileşimleri teşvik edebilir. Ortaya çıkan 3D bioprinting teknoloji heterojen TME taklit modeller oluşturmak için olanak sağlar.
3D bioprinting hızlı prototipleme türetilir ve yaşam karmaşıklığı bazılarına taklit yeteneği olan 3D microstructures imalatı sağlar doku örnekleri6,7. Geçerli bioprinting yöntemleri mürekkep püskürtmeli, ekstrüzyon ve yazdırma8lazer destekli içerir. Bunlar arasında Ekstrüzyon yöntemi heterojenite içinde yazılı matrisler malzemelerin farklı türleri farklı başlangıç yerlerde tam konumlandırma tarafından denetlenmesini sağlar. Bu nedenle, bu hücre veya matrisler birden çok türü içeren türdeş olmayan vitro modelleri imal etmek en iyi yaklaşımdır. Ekstrüzyon bioprinting kulak çevresi şeklindeki iskele9, vasküler yapılar10,11,12, kurmak ve doku13yüksek yazdırma sadakat ve hücre ile sonuçlanan, cilt için başarılı bir şekilde kullanılmıştır canlılığı. Teknoloji de çok yönlü malzeme seçimleri, bir bilinen yoğunluğu ve yüksek tekrarlanabilirlik14,15,16,17 ile gömülü hücrelerle malzemeleri yatırmak için yetenek özellikleri . Doğal ve sentetik hydrogels sık onların biyouyumluluk, bioactivity ve yapısal olarak ECM7,18 benzemeye mühendislik hidrofilik ağlarına nedeniyle 3D bioprinting için bioinks kullanılır ,19,20,21,22,23. Hydrogels, aynı zamanda beri hücreleri, yapı elemanları, geçirgenliği besin ve gazlar için yapıştırıcı siteleri içerebilir ve teşvik etmek için uygun mekanik özellikleri geliştirme24hücre avantajlı vardır. Örneğin, kollajen hydrogels integrin hücreleri matrisin için eklemek için kullanabilirsiniz anchorage siteleri sunar. Jelatin, denatüre kollajen, benzer hücre adezyon siteleri korur. Buna ek olarak, aljinat bioinert ancak Glossar divalent iyonları25,26,27,28ile oluşturarak mekanik bütünlük sağlar.
Bu çalışmada, aljinat ve jelatin, bir yerel tümör stroma mikroskobik mimarisini benzerlikler ile oluşan bir bioink olarak bileşik hidrojel geliştirdik. Meme kanseri hücreleri ve fibroblast hydrogels içinde gömülü ve üzerinden bir ekstrüzyon tabanlı bioprinter in vivo microenvironment taklit eden bir 3D model oluşturmak için yazdırılabilir. Mühendislik 3D çevre çok hücreli tümör pulcuklarının (MCTS) oluşturmak kanser hücrelerinin hücre kültürü (> 30 gün) uzun süre ile yüksek bir canlılık verir. Bu iletişim kuralı bileşik hydrogels sentezleme, malzemelerin mikroyapı ve baskıya, bioprinting hücresel türdeş olmayan modelleri, karakterize ve MCTS oluşumu gözlemleyerek yöntemleri gösterir. Bu metodolojileri ekstrüzyon bioprinting de türdeş olmayan doku modelleri farklı tasarımlar olarak içinde diğer bioinks uyuşturucu tarama, hücre göç deneyleri ve temel cep telefonundan odak çalışmaları potansiyel uygulamalar ile uygulanabilir fizyolojik fonksiyonları.

<table border="0" cellpadding="0" cellspacing="0" width="925">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Sodium alginate</td>
			<td style="width:213px;">FMC BioPolymer</td>
			<td style="width:173px;">CAS-No: 9005-38-3</td>
			<td style="width:167px;">Protanal LF 10/60 FT</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Gelatin</td>
			<td style="width:213px;">Sigma-Aldrich</td>
			<td style="width:173px;">G9391</td>
			<td>Type B gelatin from bovine skin</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Dubelcco&#39;s phosphate buffered saline (DPBS 1X)</td>
			<td style="width:213px;">Gibco</td>
			<td style="width:173px;">LS14190136</td>
			<td>1&#215;, w/o calcium, w/o magnesium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Magnetic hotplate</td>
			<td style="width:213px;">Corning&#160;</td>
			<td style="width:173px;">N/A</td>
			<td>Stirrer/hot plate model PC-420</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">50 mL centrifuge tubes</td>
			<td style="width:213px;">Corning</td>
			<td style="width:173px;">352098</td>
			<td>Falcon&#174; 50mL High Clarity PP Centrifuge Tube, Conical Bottom, Sterile</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Centrifuge</td>
			<td style="width:213px;">GMI</td>
			<td style="width:173px;">N/A</td>
			<td>Sorvall RT6000D, GMI, USA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Calcium chloride anhydrous</td>
			<td style="width:213px;">Sigma-Aldrich</td>
			<td>C1016</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">MilliQ water</td>
			<td style="width:213px;">Millipore</td>
			<td style="width:173px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Millipore 0.22 &#181;m filters</td>
			<td style="width:213px;">Millipore</td>
			<td style="width:173px;">SLGS033SB</td>
			<td>Millex-GS Syringe Filter Unit, 0.22&#160;&#181;m, mixed cellulose esters, 33&#160;mm, ethylene oxide sterilized</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Oscillation rheometer MCR 302</td>
			<td style="width:213px;">Anton Paar</td>
			<td style="width:173px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Rheometer measuring tool CP25</td>
			<td style="width:213px;">Anton Paar</td>
			<td style="width:173px;">79038</td>
			<td>Conical plate geometry for rheometer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">RheoCompass</td>
			<td style="width:213px;">Anton Paar</td>
			<td style="width:173px;">N/A</td>
			<td>Software controlling rheometer MCR 302</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Scanning electron microscope</td>
			<td style="width:213px;">Hitachi</td>
			<td style="width:173px;">N/A</td>
			<td>SEM, Hitachi SU-3500 Variable Pressure</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Paraformaldehyde, 96%, extra pure</td>
			<td style="width:213px;">Acros Organics</td>
			<td style="width:173px;">416785000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Dulbecco modified eagle medium (DMEM)</td>
			<td style="width:213px;">Gibco</td>
			<td>11965092</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Antibiotic/Antimycotic solution (100X) stabilized</td>
			<td style="width:213px;">Sigma</td>
			<td>A5955</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fetal bovine serum</td>
			<td>Wisent Bioproducts</td>
			<td style="width:173px;">080-150</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:372px;">Cell culture T-75 flasks</td>
			<td style="width:213px;">Sigma-Aldrich</td>
			<td style="width:173px;">CLS430641</td>
			<td>75 cm2 TC-Treated surface treatment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3D bioprinter BioScaffolder 3.1</td>
			<td style="width:213px;">GeSiM</td>
			<td style="width:173px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">GeSim software</td>
			<td style="width:213px;">GeSiM</td>
			<td style="width:173px;">N/A</td>
			<td>Software controlling BioScaffolder 3.1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10cc cartridge UV resist</td>
			<td>EFD Nordson</td>
			<td>7012126</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">End cap</td>
			<td>EFD Nordson</td>
			<td>7014472</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tip cap</td>
			<td>EFD Nordson</td>
			<td>7014469</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Piston&#160;</td>
			<td>EFD Nordson</td>
			<td>7012182</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stainless nozzle G25</td>
			<td>EFD Nordson</td>
			<td>7018345</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Water bath</td>
			<td style="width:213px;">VWR</td>
			<td style="width:173px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Agarose</td>
			<td style="width:213px;">Sigma-Aldrich</td>
			<td style="width:173px;">A9539</td>
			<td>Bioreagent, for molecular biology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Costar 6-well plates&#160;</td>
			<td style="width:213px;">Corning</td>
			<td style="width:173px;">3516</td>
			<td>TC-Treated Multiple Well Plates, Individually Wrapped, Sterile&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Confocal spinning disk inverted microscope</td>
			<td style="width:213px;">Olympus Life Science</td>
			<td style="width:173px;">N/A</td>
			<td>Olympus IX83</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">MTS assay kit</td>
			<td style="width:213px;">Promega</td>
			<td style="width:173px;">G3582</td>
			<td>CellTiter 96&#174; AQueous&#160;One Solution Cell Proliferation Assay&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Live/Dead viability cytotoxicity kit</td>
			<td>Molecular Probes,ThermoFisher Scientific</td>
			<td>L3224</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Trypsin 0.25/EDTA 1X</td>
			<td>Gibco</td>
			<td>25200-072</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Corning 96-well plate</td>
			<td style="width:213px;">Corning</td>
			<td style="width:173px;">3595</td>
			<td>Clear Flat Bottom Polystyrene TC-Treated Microplate, Individually Wrapped, with Low Evaporation Lid, Sterile</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Autoclave Tuttnauer</td>
			<td style="width:213px;">Heidolph Brinkmann</td>
			<td style="width:173px;">N/A</td>
			<td>Heidolph Tuttnauer 2540E Autoclave Sterilizer Electronic Model with 4 Stainless Steel Trays, 23L Capacity</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Trypan blue</td>
			<td style="width:213px;">Invitrogen&#160;</td>
			<td style="width:173px;">T10282</td>
			<td>0.4% solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Ethanol</td>
			<td style="width:213px;">Commercial Alcohols</td>
			<td style="width:173px;">P016EA95</td>
			<td>Greenfield Speciality Alcohols</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:372px;">CO2 Incubator</td>
			<td style="width:213px;">Panasonic</td>
			<td style="width:173px;">N/A</td>
			<td>MCO 19AIC-PA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">Lyophilizer&#160;</td>
			<td style="width:213px;">SP Scientific</td>
			<td style="width:173px;">N/A</td>
			<td>Virtis Sentry 2.0</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">SolidWorks</td>
			<td style="width:213px;">Dassault Systems</td>
			<td style="width:173px;">N/A</td>
			<td>A CAD software used to build demostrative propeller-like model</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:372px;">MATLAB</td>
			<td style="width:213px;">The MathWorks</td>
			<td style="width:173px;">N/A</td>
			<td>A programming software used to generate G-code for BioScaffolder 3.1</td>
		</tr>
	</tbody>
</table>

bioprintable alginate/gelatin hydrogel 3d in vitro model systems induce cell spheroid formation

Yerel tümör microenvironment hücresel, Biyokimya ve biyofizik heterojen geleneksel iki boyutlu (2D) hücre kültürü kullanarak büyüyen ölümsüzleştirdi kanser hücre hatları ile recapitulated değil. Sayede hücreler farklı türde katıştırılmış türdeş olmayan üç boyutlu (3D) tümör modelleri oluşturmak için bioprinting teknikleri kullanarak bu zorlukların üstesinden gelebilir. Aljinat ve jelatin bioprinting biyouyumluluk, Biyomimikri ve mekanik özellikleri nedeniyle istihdam en yaygın Biyomalzeme ikisidir. İki polimerler birleştirerek, bioprintable bileşik hidrojel yerli tümör stroma mikroskobik mimariye benzerlikler ile elde etti. Biz yolu ile bileşik hidrojel Reolojisi baskıya okudu ve en uygun yazdırma penceresi elde. Meme kanseri hücreleri ve fibroblast hydrogels içinde gömülü ve in vivo microenvironment taklit eden bir 3D model oluşturmak için yazdırılabilir. Bioprinted heterojen modeli uzun vadeli hücre kültürü (> 30 gün) için yüksek bir canlılık sağlar ve destekler çok hücreli tümör pulcuklarının (MCTS) içine meme kanser hücrelerinin kendinden montajlı. Biz göç ve kanser ilişkili fibroblast hücre (restorantlar) etkileşim MCTS bu modelde gözlenen. Bioprinted hücre kültür platformlar ortak kültür sistemleri olarak kullanarak, tumorigenesis bağımlılığını stroma kompozisyon çalışması için benzersiz bir araç sunar. Bu teknik özellikleri yüksek üretilen iş, düşük maliyetli ve yüksek tekrarlanabilirlik ve ayrıca geleneksel hücre monolayer kültürleri için alternatif bir model ve kanser biyoloji çalışmaya hayvan tümör modelleri sağlayabilir.

Sıcaklık süpürme A3G7 habercisi 25 ° C ve 37 ° c de ayrı bir fark gösterir Habercisi 37 ° C'de sıvı ve 1938.1 ± 84.0 mPa karmaşık bir viskozite vardır bir büyük G tarafından doğrulanır s x "G üzerinde '. Sıcaklık azaldıkça habercisi jelatin molekülleri spontan fiziksel Dolaşıklık tri-helix oluşumu29,30içine nedeniyle fiziksel jelleşme uğrar. Her iki G' ve G "artırmak ve sol-jel geçiş gösteren 3...

Watch this Scientific Journal Video about Bioprintable Alginate/Gelatin Hydrogel 3D In Vitro Model Systems Induce Cell Spheroid Formation at JoVE.com

Bioprintable Alginate/Gelatin Hydrogel 3D In Vitro Model Systems Induce Cell Spheroid Formation

Biz bir bioprintable aljinat/jelatin bioink gömülü ölümsüzleştirdi tümör ve fibroblast hücreleri içeren türdeş olmayan meme kanseri modeli geliştirilmiştir. Model vivo içinde tümör microenvironment recapitulates ve çok hücreli tümör pulcuklarının, kavrama tumorigenesis sürüş mekanizmaları içine verimli oluşumunu kolaylaştırır.

Bioprintable aljinat/jelatin hidrojel 3D Vitro Model sistemleri hücre küresel oluşumu teşvik

The cellular, biochemical, and biophysical heterogeneity of the native tumor microenvironment is not recapitulated by growing immortalized cancer cell ...

Research

JoVE Journal

Bioengineering

Bioprintable Alginate/Gelatin Hydrogel 3D In Vitro Model Systems Induce Cell Spheroid Formation

Eye Irritation Test (EIT) for Hazard Identification of Eye Irritating Chemicals using Reconstructed Human Cornea-like Epithelial (RhCE) Tissue Model

Eye Irritation Test EIT for Hazard Identification of Eye Irritating Chemicals using Reconstructed Human Cornea-like Epithelial RhCE Tissue Model

Yeniden İnşa İnsan Kornea gibi Epitelyal (RHCE) Doku Modeli kullanılarak Göz tahriş Kimyasalların Tehlike Tanımlaması Göz tahrişi Testi (EIT)

To comply with the Seventh Amendment to the EU Cosmetics Directive and EU REACH legislation, validated non-animal alternative methods for reliable and ...

Modeling Ovarian Cancer Multicellular Spheroid Behavior in a Dynamic 3D Peritoneal Microdevice

Dinamik 3D Periton microdevice Modelleme Yumurtalık Kanseri Multisellüler Sfero Davranış

Ovarian cancer is characterized by extensive peritoneal metastasis, with tumor spheres commonly found in the malignant ascites. This is associated with ...

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

bir<em&gt; İn Vitro</emKemirgen İntervertebral Disk&gt; Organ Kültürü Modeli

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and ...

Improved 3D Hydrogel Cultures of Primary Glial Cells for In Vitro Modelling of Neuroinflammation

Improved 3D Hydrogel Cultures of Primary Glial Cells for In Vitro Modelling of Neuroinflammation

Geliştirilmiş 3D hidrojel Neuroinflammation Vitro modelleme için birincil gliyal hücreler kültürleri

In the central nervous system, numerous acute injuries and neurodegenerative disorders, as well as implanted devices or biomaterials engineered to enhance ...

3D Printed Porous Cellulose Nanocomposite Hydrogel Scaffolds

3D gözenekli selüloz Nanocomposite hidrojel iskele baskılı

This work demonstrates the use of three-dimensional (3D) printing to produce porous cubic scaffolds using cellulose nanocomposite hydrogel ink, with ...

A Human 3D Extracellular Matrix-Adipocyte Culture Model for Studying Matrix-Cell Metabolic Crosstalk

Bir İnsan 3D Ekstrasellüler Matrix-Adiposit Kültür Modeli Matrix-Hücre Metabolik Crosstalk Çalışma için

The extracellular matrix (ECM) plays a central role in regulating tissue homeostasis, engaging in crosstalk with cells and regulating multiple aspects of ...

Protocols of 3D Bioprinting of Gelatin Methacryloyl Hydrogel Based Bioinks

Jelatin Metacryloyl Hidrojel Bazlı Bioinks 3D Biyobaskı Protokolleri

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

Using Multilayered Hydrogel Bioink in Three-Dimensional Bioprinting for Homogeneous Cell Distribution

Homojen Hücre Dağılımı için Üç Boyutlu Biyobaskıda Çok Katmanlı Hidrojel Bioink'in Kullanılması

During the extrusion-based three-dimensional bioprinting process, liquid-like bioinks with low viscosity can protect cells from membrane damage induced by ...

Advanced 3D Liver Models for In vitro Genotoxicity Testing Following Long-Term Nanomaterial Exposure

Uzun Süreli Nanomalzeme Maruziyetini Takiben İn vitro Genotoksisite Testi için Gelişmiş 3D Karaciğer Modelleri

Due to the rapid development and implementation of a diverse array of engineered nanomaterials (ENM), exposure to ENM is inevitable and the development of ...

3D Printing of In Vitro Hydrogel Microcarriers by Alternating Viscous-Inertial Force Jetting

3D Printing of In Vitro Hydrogel Microcarriers by Alternating Viscous-Inertial Force Jetting

Alternatif Viskoz-Atalet Kuvveti Jetting ile In Vitro Hydrogel Mikrokaçörlerinin 3D Baskısı

Microcarriers are beads with a diameter of 60-250 &#181;m and a large specific surface area, which are commonly used as carriers for large-scale cell ...