Name: tissue engineering of tumor stromal microenvironment with application to cancer cell invasion
Uploaded: 2014-03-18T15:00:00
Description: Watch this Scientific Journal Video about Tissue Engineering of Tumor Stromal Microenvironment with Application to Cancer Cell Invasion at JoVE.com

APS er støttet af Debra International og British Skin Foundation. YZN understøttes af A * STAR - University of Dundee Partnership ph.d.-studiet.

Denne undersøgelse blev udført i overensstemmelse med Helsinki-deklarationen Principper og blev godkendt af de relevante etiske komitéer. 1.. Udarbejdelse af medier og reagenser <ol><li> Fremstilling af 200x af L-ascorbinsyre-2-phosphat-lager <ol><li> 29 mg L-ascorbinsyre 2-phosphat pr 5 ml Dulbeccos Modified Eagle Medium (DMEM) opløses og filtreres gennem 0,22 um membranfilter. Store som 0,25 ml sterile portioner i -20 ° C. </li><li> Tilføj 0,25 ml alikvot af 200x L-ascorbinsyre 2-p...

<ol>
	<li>Bell, E., Ehrlich, H. P., Buttle, D. J., Nakatsuji, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Living+tissue+formed+in+vitro+and+accepted+as+skin-equivalent+tissue+of+full+thickness.">Living tissue formed in vitro and accepted as skin-equivalent tissue of full thickness.</a> Science. 211 (4486), 1052-1054 (1981).</li><li>Asselineau, D., Bernhard, B., Bailly, C., Darmon, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidermal+morphogenesis+and+induction+of+the+67+kD+keratin+polypeptide+by+culture+of+human+keratinocytes+at+the+liquid-air+interface.">Epidermal morphogenesis and induction of the 67 kD keratin polypeptide by culture of human keratinocytes at the liquid-air interface.</a> Exp. Cell Res. 159 (2), 536-539 (1985).</li><li>Pruni&#233;ras, M. M., R&#233;gnier, M. M., Woodley, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+Cultivation+of+Keratinocytes+with+an+Air-Liquid+Interface.">Methods for Cultivation of Keratinocytes with an Air-Liquid Interface.</a> J. Invest. Dermatol. 81 (1 Suppl), 28-33 (1983).</li><li>Prunieras, M., R&#233;gnier, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+procedure+for+culturing+human+epidermal+cells+on+allogenic+or+xenogenic+skin:+preparation+of+recombined+grafts.">New procedure for culturing human epidermal cells on allogenic or xenogenic skin: preparation of recombined grafts.</a> Ann. Chir. Plast. 24 (4), 357-362 (1979).</li><li>Gaggioli, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibroblast-led+collective+invasion+of+carcinoma+cells+with+differing+roles+for+RhoGTPases+in+leading+and+following+cells.">Fibroblast-led collective invasion of carcinoma cells with differing roles for RhoGTPases in leading and following cells.</a> Nature. 9 (12), 1392-1400 (2007).</li><li>Bhowmick, N. A., Neilson, E. G., Moses, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stromal+fibroblasts+in+cancer+initiation+and+progression.">Stromal fibroblasts in cancer initiation and progression.</a> Nature. 432 (7015), 332-337 (2004).</li><li>Erez, N., Truitt, M., Olson, P., Hanahan, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer-Associated+Fibroblasts+Are+Activated+in+Incipient+Neoplasia+to+Orchestrate+Tumor-Promoting+Inflammation+in+an+NF-&#954;B-Dependent.">Cancer-Associated Fibroblasts Are Activated in Incipient Neoplasia to Orchestrate Tumor-Promoting Inflammation in an NF-&#954;B-Dependent.</a> Manner. Cancer Cell. 17 (2), 135-147 (2010).</li><li>Harkness, R. D., Marko, A. M., Muir, H. M., Neuberger, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+metabolism+of+collagen+and+other+proteins+of+the+skin+of+rabbits.">The metabolism of collagen and other proteins of the skin of rabbits.</a> Biochem. J. 56 (4), 558-569 (1954).</li><li>Ehrmann, R. L., Gey, G. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+growth+of+cells+on+a+transparent+gel+of+reconstituted+rat-tail+collagen.">The growth of cells on a transparent gel of reconstituted rat-tail collagen.</a> J. Natl. Cancer Inst. 16 (6), 1375-1403 (1956).</li><li>Kleinman, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Basement+membrane+complexes+with+biological+activity.">Basement membrane complexes with biological activity.</a> Biochemistry. 25 (2), 312-318 (1986).</li><li>Vukicevic, S., Kleinman, H. K., Luyten, F. P., Roberts, A. B., Roche, N. S., Reddi, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+multiple+active+growth+factors+in+basement+membrane+Matrigel+suggests+caution+in+interpretation+of+cellular+activity+related+to+extracellular+matrix+components.">Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation of cellular activity related to extracellular matrix components.</a> Exp. Cell Res. 202 (1), 1-8 (1992).</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=BD+Biosciences+-+Discover+Labware.">BD Biosciences - Discover Labware.</a> SPC-356230 Rev 5.0 at Rev 5.0. , .</li><li>Hughes, C. S., Postovit, L. M., Lajoie, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrigel:+a+complex+protein+mixture+required+for+optimal+growth+of+cell+culture.">Matrigel: a complex protein mixture required for optimal growth of cell culture.</a> Proteomics. 10 (9), 1886-1890 (2010).</li><li>Ng, Y. Z., Dayal, J. H., South, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+Predisposition+to+Cutaneous+Squamous+Cell+Carcinoma.">Genetic Predisposition to Cutaneous Squamous Cell Carcinoma.</a> , .</li><li>Christiano, A. M. A., Greenspan, D. S. D., Lee, S. S., Uitto, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cloning+of+human+type+VII+collagen.+Complete+primary+sequence+of+the+alpha+1(VII)+chain+and+identification+of+intragenic+polymorphisms.">Cloning of human type VII collagen. Complete primary sequence of the alpha 1(VII) chain and identification of intragenic polymorphisms.</a> J. Biol. Chem. 269 (32), 20256-20262 (1994).</li><li>Hovnanian, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+linkage+of+recessive+dystrophic+epidermolysis+bullosa+to+the+type+VII+collagen+gene.">Genetic linkage of recessive dystrophic epidermolysis bullosa to the type VII collagen gene.</a> J. Clin. Invest. 90 (3), 1032-1036 (1992).</li><li>Ryyn&#228;nen, M. M., Ryyn&#228;nen, J. J., Sollberg, S. S., Iozzo, R. V. R., Knowlton, R. G. R., Uitto, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+linkage+of+type+VII+collagen+(COL7A1)+to+dominant+dystrophic+epidermolysis+bullosa+in+families+with+abnormal+anchoring+fibrils.">Genetic linkage of type VII collagen (COL7A1) to dominant dystrophic epidermolysis bullosa in families with abnormal anchoring fibrils.</a> J. Clin. Invest. 89 (3), 974-980 (1992).</li><li>Ng, Y. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibroblast-derived+dermal+matrix+drives+development+of+aggressive+cutaneous+squamous+cell+carcinoma+in+patients+with+recessive+dystrophic+epidermolysis+bullosa.">Fibroblast-derived dermal matrix drives development of aggressive cutaneous squamous cell carcinoma in patients with recessive dystrophic epidermolysis bullosa.</a> Cancer Res. 72 (14), 3522-3534 (2012).</li><li>Larouche, D., Paquet, C., Fradette, J., Carrier, P., Auger, F. A., Germain, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+15.">Chapter 15.</a> Methods Mol. Biol. 482, 233-256 (2009).</li><li>Pouliot, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reconstructed+human+skin+produced+in+vitro+and+grafted+on+athymic+mice).">Reconstructed human skin produced in vitro and grafted on athymic mice).</a> Transplantation. 73 (11), 1751-1757 (2002).</li><li>Purdie, K. J., Pourreyron, C., South, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+culture+of+squamous+cell+carcinoma+lines.">Isolation and culture of squamous cell carcinoma lines.</a> Methods Biol. 731, 151-159 (2011).</li><li>Pinnell, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+collagen+biosynthesis+by+ascorbic+acid:+a+review.">Regulation of collagen biosynthesis by ascorbic acid: a review.</a> Yale J. Biol. Med. 58 (6), 553-559 (1985).</li><li>Hata, R., Senoo, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=L-ascorbic+acid+2-phosphate+stimulates+collagen+accumulation,+cell+proliferation,+and+formation+of+a+three-dimensional+tissuelike+substance+by+skin+fibroblasts.">L-ascorbic acid 2-phosphate stimulates collagen accumulation, cell proliferation, and formation of a three-dimensional tissuelike substance by skin fibroblasts.</a> J. Cell. Physiol. 138 (1), 8-16 (1988).</li></ol>

På grund af karakteren af ​​dette forsøg, kan den samlede tid, der kræves for færdiggørelse være op til to måneder. I hele denne gang, skal yderste omhu og sterile vævskultur praksis anvendes til at forhindre mikrobiel kontaminering. Bortset fra sin rolle som en cofaktor i syntesen af hydroxyprolin og hydroxylysin af kollagener, ascorbinsyre stimulerer collagen specifik mRNA-ekspression i fibroblaster 22. Ascorbinsyre valg her er den mere st...

Anvendelse af biomateriale i 3D vævskultur har gjort det muligt for forskerne at studere celle adfærd i laboratoriet under fysiologiske betingelser mere beslægtet med et in vivo-miljøet end én gentaget med 2D-adhæsion og et plastunderlag. Især fremad store fremskridt der er gjort i modellering lagdelt epiteler med vedtagelsen af 3D dyrkningsmetoder på luft-væske grænsefladen 1-4. Sådanne teknikker trofast efterligner keratinocytdifferentiering og tumorcelleinvasion muliggør større fleksibilitet og troskab for forskere studerer disse processer. Valget af biomateriale substrat til at efterligne det stromale miljø har primært involveret anvendelse af type I collagen, Engelbreth-Holm-Swarm mus sarkom matrix og de-epidermized dermis. For eksempel har cancerassocierede fibroblaster vist sig at bidrage mod cancer invasion 5, initiering og progression via stromale-epitel interaktioner 6,7 when dyrkes i sådanne substrater. Den gyldne standard for at efterligne det stromale miljø i huden, de største og mest udbredte studerede stratificerede epitel ved hjælp af sådanne teknikker, anses for at være de-epidermized menneskelige dermis (DED). Udarbejdelse af DED omfatter fjernelse af overhuden via trypsinisering eller fysisk afstandtagen fra humant kadaverhud 3,4. Dog kan adgang til en sådan hud være meget svært for laboratorierne ikke er associeret med kliniske institutioner, og syge dermis er nær umuligt at opnå. Som et alternativ, laboratorier ofte bruger en kombination af type I kollagen (isoleret fra rotte haler) og / eller Engelbreth-Holm-Swarm mus sarkom matrix. Efter opdagelsen i 1927 af Nageotte 8 at kollagen let kan isoleres under anvendelse af eddikesyre og saltudfældning, blev dets anvendelse på vævskultur senere udviklet af Huzella og kolleger 9.. Kollagen overtræk viste sig at være overlegen i forhold til glas til cellekultur af 29 stammer og vævseksplantater som interrogerede af Ehrmann og Gey 9. I øjeblikket er den største type af kollagen anvendes i vævskultur isoleret fra rotte-hale sener, og er normalt købt fra kommercielle kilder. Den ulempe til trofast substrat sammenfatning er imidlertid, at rotte hale kollagen er ikke identisk med human collagen, eller de humane dermis, hvor type I og III collagens er til stede som hovedbestanddele, og isolerede rotte hale kollagen er uvægerligt fragmenteret. Engelbreth-Holm-Swarm mus sarkom matrix er en gelatinøs proteinblanding udskilles af dyrkede Engelbreth-Holm-Swarm muse sarcomaceller 10. De vigtigste bestanddele er laminin, type IV collagen, heparinsulfat proteoglycan entactin og nidogen og den nøjagtige mængde af disse proteiner vil variere fra batch til batch. Bortset fra strukturelle proproteiner, indeholder denne matrix også betydelige niveauer af vækstfaktorer, såsom transformerende vækstfaktor β, epidermal vækstfaktor, insulin-lignende vækst faktor 1, kvæg fibroblast-vækstfaktor, og trombocyt-afledt vækst faktor, som ville ændre cellulære adfærd 11,12. Peger mod det store kompleksitet Engelbreth-Holm-Swarm mus sarkom matrix, blev identificeret i alt 1.851 proteiner i en nylig proteom undersøgelse 13. I lyset af de rige og komplekse karakter af denne matrix, er forsigtighed blevet rådgivet ved fortolkning og sammenligning af forskellige eksperimenter med brug af det 11. Vores laboratorier har en stor interesse i genetiske hudsygdomme, især dem med en disposition for at udvikle kutant planocellulært karcinom (CSCC) 14.. I tilfælde af recessive dystrofisk epidermolysis bullosa (RDEB), en alvorlig blærer sygdom med germline mutationer i COL7A1-genet <sop&gt; 15-17, har vi fastslået, at den dermale mikromiljø i disse patienter er tumorfremmende 18. I løbet af denne undersøgelse, vi var ude af stand til at vurdere tumorfremmende egenskaber dermalfibroblaster indlejret i kollagen I / Engelbreth-Holm-Swarm mus sarkom matrix og undersøgte metoder til vurdering cellernes egne, indfødte matrix. For at opnå dette, har vi ændret en tidligere teknik fra laboratoriet af Lucie Germain arbejder på menneskers hud ækvivalenter 19,20. Germain teknik var i stand til at rekonstruere den menneskelige hud med velorganiseret basalmembran med primære humane keratinocytkulturer og fibroblastkulturer i mangel af en syntetisk eller cadaveric stillads. I dette papir de trin, der anvendes til rekapitulere den kutane tumor stroma mikromiljø (native matrix) afledt direkte fra primære stromale fibroblaster in vitro beskrives 18. Native matricer produced af langvarig kultur af fibroblaster blev anvendt som svarer til assay for CSCC celleinvasion dermal. Vi præsenterer data ved hjælp af indfødte matrix stammer fra enten den ekstracellulære matrix, der udskilles af RDEB fibroblaster (med mangelfuld typen VII kollagen (C7)) eller fra RDEB fibroblaster retrovirustransformerede transduceret med en type VII collagen udtrykker konstruere og demonstrere dybtgående effekt af en enkelt collagen på tumor celleinvasion.

<table border="0" cellpadding="0" cellspacing="0" width="352">
	<colgroup>
		<col span="4" style="width:66pt" width="88" />
	</colgroup>
	<tbody>
		<tr height="64" style="height:48.0pt">
			<td class="xl63" height="64" style="height:48.0pt;width:66pt" width="88">L-Ascorbic acid 2-phosphate</td>
			<td class="xl63" style="width:66pt" width="88">Sigma</td>
			<td class="xl63" style="width:66pt" width="88">A8960</td>
			<td class="xl64" style="width:66pt" width="88"></td>
		</tr>
		<tr height="42" style="height:31.5pt">
			<td class="xl65" height="42" style="height:31.5pt;width:66pt" width="88">DMEM with l-glutamine,</td>
			<td class="xl67" rowspan="2" style="border-bottom:1.0pt solid black;
 border-top:none;width:66pt" width="88">Life Technologies</td>
			<td class="xl67" rowspan="2" style="border-bottom:1.0pt solid black;
 border-top:none;width:66pt" width="88">11995-073</td>
			<td class="xl67" rowspan="2" style="border-bottom:1.0pt solid black;
 border-top:none;width:66pt" width="88"></td>
		</tr>
		<tr height="106" style="height:79.5pt">
			<td class="xl63" height="106" style="height:79.5pt;width:66pt" width="88">4,500 mg/L d-glucose, 110 mg/L sodium pyruvate</td>
		</tr>
		<tr height="85" style="height:63.75pt">
			<td class="xl63" height="85" style="height:63.75pt;width:66pt" width="88">100x Penicillin-Streptomycin</td>
			<td class="xl63" style="width:66pt" width="88">Life Technologies</td>
			<td class="xl63" style="width:66pt" width="88">15070</td>
			<td class="xl64" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl63" height="43" style="height:32.25pt;width:66pt" width="88">Vaseline</td>
			<td class="xl63" style="width:66pt" width="88">VWR</td>
			<td class="xl63" style="width:66pt" width="88">PROL28908.290</td>
			<td class="xl64" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl63" height="43" style="height:32.25pt;width:66pt" width="88">Clonal cylinders</td>
			<td class="xl63" style="width:66pt" width="88">Sigma</td>
			<td class="xl63" style="width:66pt" width="88">Z370789</td>
			<td class="xl64" style="width:66pt" width="88"></td>
		</tr>
		<tr height="148" style="height:111.0pt">
			<td class="xl63" height="148" style="height:111.0pt;width:66pt" width="88">Nylon Net Filter Disc Hydrophilic 100um 25um diameter 100/pk</td>
			<td class="xl63" style="width:66pt" width="88">Millipore</td>
			<td class="xl63" style="width:66pt" width="88">NY1H02500</td>
			<td class="xl64" style="width:66pt" width="88"></td>
		</tr>
		<tr height="127" style="height:95.25pt">
			<td class="xl63" height="127" style="height:95.25pt;width:66pt" width="88">Bent stainless steel wire mesh support</td>
			<td class="xl63" style="width:66pt" width="88">Made in house</td>
			<td class="xl63" style="width:66pt" width="88"></td>
			<td class="xl66" style="width:66pt" width="88">Dimensions were made so that the mesh would fit into 6-well plates</td>
		</tr>
	</tbody>
</table>

tissue engineering of tumor stromal microenvironment with application to cancer cell invasion

3D organotypiske kulturer af epitelceller på en matrix indlejret med mesenchymale celler er almindeligt anvendt til at studere epitelial celledifferentiering og invasion. Rotte hale kollagen type I og / eller matrix afledt fra Engelbreth-Holm-Swarm muse sarcomaceller traditionelt er blevet anvendt som substrater til at modellere matrix eller stromale mikromiljø, hvori mesenchymale celler (sædvanligvis fibroblaster) er befolket. Selvom forsøg med sådanne matricer er meget informativ, kan det hævdes, at på grund af en mere tungtvejende tilstedeværelsen af ​​en enkelt protein (som i type I kollagen), eller et højt indhold af basalmembrankomponenter og vækstfaktorer (såsom i matrix stammer fra mus sarcomaceller), har disse substrater ikke bedst afspejler bidraget til matrixsammensætning fra stromacellerne selv. At studere indfødte matricer produceret af primær dermalfibroblaster isoleret fra patienter med en tumor udsat, genetisk blærer lidelse (recessive dystrofiskepidermolysis bullosa), har vi tilpasset en eksisterende native matrix protokol til at studere tumorcelleinvasion. Fibroblaster tilskyndet til at producere deres egen matrix over en længere periode i kultur. Denne native matrix derefter løsrevet fra dyrkningsskålen og epitelceller udsås på det før hele cokultur hæves til luft-væske-grænsefladen. Cellulær differentiering og / eller invasion kan derefter vurderes over tid. Denne teknik giver mulighed for at vurdere epitel-mesenchymale celle interaktioner i en 3D-indstilling uden behov for en syntetisk eller udenlandsk matrix med den eneste ulempe er den lange periode, der kræves for at producere den native matrix. Her beskrives anvendelsen af ​​denne teknik til at vurdere evnen af ​​et enkelt molekyle udtrykt af fibroblaster, type VII collagen, til at inhibere tumorcelleinvasion.

<html> Denne teknik åbner mulighed for at undersøge og sammenligne den invasive opførsel af tumorceller (i dette tilfælde CSCC) under forskellige 3D stromale miljøer. Ved hjælp af denne teknik, kan ikke blot C7-deficiente native matricer, gentaget den RDEB dermal miljø dannes, men også yderligere matricer, som er blevet genetisk manipuleret til over-express C7 18. Som det ses i figur 2, invasion af RDEB CSCC keratinocytter var signifikant forsinket i C7-overudtrykker native matricer i ...

Watch this Scientific Journal Video about Tissue Engineering of Tumor Stromal Microenvironment with Application to Cancer Cell Invasion at JoVE.com

Tissue Engineering of Tumor Stromal Microenvironment with Application to Cancer Cell Invasion

Tissue Engineering af Tumor Stromal mikromiljø med Application til cancercelleinvasion

Manipuleret væv fibroblast-afledte native matrix er en ny værktøj til at generere en stromal substrat, som understøtter epitelcelleproliferation og differentiering. Her en protokol at anvende denne metode til at vurdere virkningerne af forskellige stromale celletyper på tumor cellebiologi præsenteres.

3D organotypic cultures of epithelial cells on a matrix embedded with mesenchymal cells are widely used to study epithelial cell differentiation and ...

Research

JoVE Journal

Bioengineering

Microfluidic Device for Recreating a Tumor Microenvironment in Vitro

Microfluidic Device for Recreating a Tumor Microenvironment in Vitro

We have developed a microfluidic device that mimics the delivery and systemic clearance of drugs to heterogeneous three-dimensional tumor tissues in ...

Directed Cellular Self-Assembly to Fabricate Cell-Derived Tissue Rings for Biomechanical Analysis and Tissue Engineering

Instrueret Cellular saml-selv at fabrikere Cell-afledt Tissue Ringe til Biomekanisk analyse og Tissue Engineering

Each year, hundreds of thousands of patients undergo coronary artery bypass surgery in the United States.1 Approximately one third of these patients do ...

Three-dimensional Cell Culture Model for Measuring the Effects of Interstitial Fluid Flow on Tumor Cell Invasion

Tredimensional Cell Culture Model for måling af virkningerne af interstitielvæske Flow på tumorcelleinvasion

The growth and progression of most solid tumors depend on the initial transformation of the cancer cells and their response to stroma-associated signaling ...

Engineering Skeletal Muscle Tissues from Murine Myoblast Progenitor Cells and Application of Electrical Stimulation

Engineering skeletmuskel Væv fra murine myoblast progenitorceller og anvendelse af elektrisk stimulering

Engineered muscle tissues can be used for several different purposes, which include the production of tissues for use as a disease model in vitro, e.g. to ...

Tissue Engineering of a Human 3D in vitro Tumor Test System

Tissue Engineering of a Human 3D in vitro Tumor Test System

Tissue Engineering af et menneske 3D<em&gt; In vitro</em&gt; Tumor Test System

Cancer is one of the leading causes of death worldwide.  Current therapeutic strategies are predominantly developed in 2D culture  systems, which ...

Engineering Fibrin-based Tissue Constructs from Myofibroblasts and Application of Constraints and Strain to Induce Cell and Collagen Reorganization

Engineering Fibrin-baserede vævskonstruktioner fra myofibroblasts og Application of Begrænsninger og stamme at fremkalde Cell og kollagen (Re) Organisation

Collagen content and organization in developing collagenous tissues can be influenced by local tissue strains and tissue constraint. Tissue engineers aim ...

Long-term Intravital Immunofluorescence Imaging of Tissue Matrix Components with Epifluorescence and Two-photon Microscopy

Langsigtet Intravital Immunofluorescens Imaging af Tissue Matrix Komponenter med epifluorescens og to-foton mikroskopi

Besides being a physical scaffold to maintain tissue morphology, the extracellular matrix (ECM) is actively involved in regulating cell and tissue ...

Concentric Gel System to Study the Biophysical Role of Matrix Microenvironment on 3D Cell Migration

Koncentrisk Gel System til Undersøgelse den biofysiske rolle Matrix mikromiljø på 3D Cell Migration

The ability of cells to migrate is crucial in a wide variety of cell functions throughout life&#160;from embryonic development and wound healing&#160;to ...

Sandwich-like Microenvironments to Harness Cell/Material Interactions

Sandwich-lignende mikromiljøer at udnytte Cell / Materiale Interactions

Cell culture has been traditionally carried out on bi-dimensional (2D) substrates where cells adhere using ventral receptors to the biomaterial surface. ...

In Vitro Model of Physiological and Pathological Blood Flow with Application to Investigations of Vascular Cell Remodeling

In Vitro Model of Physiological and Pathological Blood Flow with Application to Investigations of Vascular Cell Remodeling

In vitro model af fysiologiske og patologiske Blood Flow med Application til Undersøgelser af vaskulær Cell Remodeling

Vascular disease is a common cause of death within the United States. Herein, we present a method to examine the contribution of flow dynamics towards ...