Name: improving 2d and 3d skin in vitro models using macromolecular crowding
Uploaded: 2016-08-22T13:00:00
Description: Watch this Scientific Journal Video about Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding at JoVE.com

This work was supported by the Biomedical Research Council, Singapore through core support to the Institute of Medical Biology and grant SPF2013/005. M.R. was supported by a NUS Faculty Research Committee Grant (Engineering in Medicine) (M.R.) R-397-000-081-112, and the NUS Tissue Engineering Program (NUSTEP). The authors would like to thank Professor Irene Leigh for providing the collagen VII antibody. Electron microscopy work was carried out at the National University of Singapore Electron Microscopy Unit.

1. Makromolekylær Fortrængning i 2D hudceller Cultures <ol><li> Seed 50.000 celler (primære fibroblaster eller primære keratinocytter eller co-kultur af primære fibroblaster og keratinocytter) pr brønd af en 24-brønds plade. Seed celler i 1 ml af den tilsvarende celletype vækstmedier. </li><li> Tillad celler til at adhærere natten over, i et 37 ° C inkubator ved 5% CO2. </li><li> Kassér gamle medier og erstatte med 1 ml frisk medium indeholdende makromolekylære Crowders og 100 uM ascorbinsyre. B...

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Enhanced extracellular matrix is obtained upon introduction of macromolecular crowders to cell culture, owing to the excluded volume effect which increases the propeptide cleavage by proteinases. This results in extracellular matrix, in particular collagen, to be processed faster and deposited on the culture surface. While other groups have obtained thick fibroblast cell layers, it involved a culture time of several weeks20. In contrast, MMC described in this Protocol, dramatically shortens the culture time wh...

Huden danner en beskyttende barriere ved at forhindre vandtab og patogen post. Den består af tre hovedkomponenter; de stromale rige dermis 1, en ​​stratificeret epidermis 2,3,4 på toppen af det, og dermo-epidermal junction i mellem 5,6. Dermis består hovedsageligt af kollagen og elastiske fibre og er tyndt befolket med fibroblaster 7. I modsætning hertil er den celle-rige epidermis består af flere lag af keratinocytter. Keratinocytterne af inderste lag er proliferativ og tilvejebringe nye basale celler, som fornyer og erstatte terminalt differentierende keratinocytter, der konstant flytter til den udvendige mest lag af huden og har mistet deres kerner og cytoplasmatiske materiale, hvilket resulterer i en forhornede lag, som undergår afskalning. Den dermale-epidermale krydset, en specifik type basalmembran, er en kompleks struktur bestående af indbyrdes forbundne matrixmolekyler, der tethers epidermis til dermis. Kollagen I fibre af dermis interlace med kollagen VII forankring fibriller, som er forankret til collagen IV rige lamina densa. Forankring filamenter (laminin 5, collagen XVII og integriner) til gengæld forbinde lamina densa med hemidesmosomer af basale keratinocytter. Basale keratinocytter (stratum basale) har kapacitet til at formere sig og forny, samt differentiere og stratificere at danne suprabasallagene; stratum spinosum, stratum granulosum, og endelig stratum corneum, som repræsenterer kontakt overfladen af huden med miljøet. Undervejs fra det basale lag til forhornede lag, keratinocytter skifter ekspressionsmønstre for cytokeratiner og endelig undergår apoptose og indkapsle sig i forhornede kuverter, skrog specifikke proteiner, som er kovalent tværbundet ved transglutaminase aktivitet. Genskabelse hud og dets lag in vitro, herunder de komplekse strukturer<html> dermal-epidermal krydset og dermal ekstracellulære matrix, og at emulere cornificering proces, har længe fascineret forskere og bioengineers som en udfordrende opgave. Der har været betydelige fremskridt i huden tissue engineering, for eksempel, den vellykkede udvinding af hudceller fra patientens biopsier og generering af huden organotypiske kulturer ved hjælp patient-afledte hudceller 8. Dog forbliver uløste problemer vedrørende dårlig sekretion af ekstracellulære matrix proteiner ved hudceller selv og resulterer i sub-optimale hudmodeller. Desuden den nødvendige tid til at generere 3D organotypisk hud co-kultur ved hjælp af de nuværende protokoller varierer mellem fire til otte uger, en tidsramme, der potentielt kan afkortes med indarbejdelsen af ​​makromolekylære Crowders. Reduktion kultur tid sparer reagens omkostninger, reducerer forekomsten af ​​celle ældning og reducerer ventetiden for patienten skal produktet bruges i klinikken. t &quot;&gt; Makromolekylær fortrængning (MMC) involverer om særlige makromolekyler til dyrkningsmediet at generere ekskluderede volumen effekter. Disse påvirker enzymatiske reaktionshastigheder herunder den proteolytiske spaltning af prokollagen, der er langsomme under standard vandige dyrkningsbetingelser 9-13. Under MMC, enzymatiske reaktioner er fremskyndet uden at øge mængden af reagenser 14,15 resulterer i, i tilfælde af prokollagen spaltning, en forøget mængde af kollagen i-molekyler i overfyldte kulturer i sammenligning med Uncrowded kontroller 10. da omdannelsen af procollagen til collagen tillader dannelsen af kollagen samlinger, fibroblaster dyrket med MMC i 48 timer gav signifikant mere kollagen i sammenlignet med Uncrowded fibroblastkulturer overvåges i op til fire uger 11,16,17. Udover virkninger på enzymatiske aktiviteter, som påvirker dannelsen, stabilisering og remodellering af ECM, MMC også har vist sig at direkte forbedre og modulere kollagenfiberdannelse 18,19. Vi præsenterer her en metode til at forbedre den ekstracellulære matrix (ECM) produktion af hudceller, navnlig dermale fibroblaster og epidermale keratinocytter. Derudover viser vi, at den berigede ECM produceret under MMC i monolagskulturer kan decellulariserede og anvendt som rent celleafledt matrix (CDM). Vi bruger en ikke-konventionelle tilgang til at visualisere og fuldt ud forstår ECM deponeret af hudceller dyrket med MMC. Interferens refleksioner mikroskopi anvendes typisk til at studere celle-matrix interaktioner eller celle-til-glas kontaktpunkter. Denne teknik blev anvendt i vores system til at se den samlede matrix aflejres på glasoverfladen. Interferens refleksioner mikroskopi blev koblet med fluorescerende immunfarvning for at opnå den mest mængden af ​​information i form af ekstracellulær matrix komposition og mønster, i nærvær og fravær af MMC. Organotypic hud co-kulturer er en klassisk metode til at modellere huden in vitro i en tredimensional kontekst. Mens todimensionale co-kulturer kan give væsentlige oplysninger, er det begrænset, når oversætte disse data og anvende det tilbage til et in vivo miljø, der er i sagens natur en tredimensionel struktur. Hud keratinocytter, i særdeleshed, er polariseret og indeholder apikale og basale segmenter, som er afgørende for homeostase og celle vedhæftning. Derudover er ekspressionen af ​​typiske suprabasale proteiner i keratinocytter over det basale lag, såsom keratin 1, keratin 10 og filaggrin er kun til stede i løbet af lagdeling og terminal differentiering af keratinocytter. Som terminal differentiering er næppe til stede i typiske monolagskulturer er suprabasale proteinekspression normalt ikke nået i denne kultur-system. Derfor begynder organotypiske kulturer nedsænket i dyrkningsmediet, men derefter løftet til en luft-væske grænsefladen til at køre keratinocyte differentiering. Dette resulterer i ekspressionen af ​​stratification markører, selv horndannelse og en generelt bedre afspejling af epidermal fysiologi. Mens andre grupper tidligere har genereret organotypisk hud co-kulturer med succes, har etableringen af ​​en funktionel dermo-epidermal krydset zone været et problem. Her præsenteres en ny fremgangsmåde til dyrkning af organotypiske hud co-kulturer med en forbedret basalmembran, i sammentrængt tidsramme og uden at kompromittere modenhed af disse konstruktioner. Dette ville give huden mimetika til in vitro-modellering, studiet af huden biologi og et sortiment af screening-assays.

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Ascorbic acid</td>
			<td style="width:181px;">Wako</td>
			<td style="width:175px;">013-12061</td>
			<td style="width:499px;">Cell culture media additive</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cell culture inserts&#160; (6-well format)</td>
			<td style="width:181px;">Greiner Bio-One</td>
			<td style="width:175px;">657610</td>
			<td>Organotypic cultures</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Citrate solution</td>
			<td style="width:181px;">Dako</td>
			<td style="width:175px;">S2369</td>
			<td style="width:499px;">DakoCytomation Target Retrieval Solution Citrate, pH 6 (x10)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Collagen (rat tail)</td>
			<td style="width:181px;">Corning</td>
			<td style="width:175px;">354236</td>
			<td>Organotypic cultures</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">CnT-57&#160;</td>
			<td style="width:181px;">CELLnTEC</td>
			<td style="width:175px;">CnT-57</td>
			<td>Cell culture media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">DAB Substrate + chromogen kit</td>
			<td style="width:181px;">Dako</td>
			<td style="width:175px;">K3468</td>
			<td>Histology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Deep-well plate (6-well format)</td>
			<td style="width:181px;">Corning</td>
			<td style="width:175px;">355467</td>
			<td>Organotypic cultures</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">ECL detection reagent</td>
			<td style="width:181px;">GE Healthcare Life Sciences</td>
			<td style="width:175px;">RPN2106</td>
			<td>Amersham ECL Western Blotting Detection Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Electron microscope (TEM)</td>
			<td>&#160;JEOL&#160;</td>
			<td>&#160;JEM-1010 (100kV)</td>
			<td>Transmission electron microscopy</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Fibroblast media (FM)</td>
			<td></td>
			<td></td>
			<td>High Glucose-DMEM + 10% Fetal Bovine Serum + 1% Penicillin Streptomycin</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">Ficoll PM70</td>
			<td style="width:181px;">GE Healthcare Life Sciences</td>
			<td style="width:175px;">17-0310-05</td>
			<td>Macromolecular crowder</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">Ficoll PM400</td>
			<td style="width:181px;">GE Healthcare Life Sciences</td>
			<td style="width:175px;">17-0300-05</td>
			<td>Macromolecular crowder</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Keratinocyte serum-free media (KSFM)</td>
			<td style="width:181px;">Life Technologies</td>
			<td style="width:175px;">17005-042</td>
			<td>Cell culture media</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">Lysis buffer</td>
			<td style="width:181px;">ThermoFisher Scientific</td>
			<td style="width:175px;">89900</td>
			<td style="width:499px;">Lysis buffer for protein extraction. A protease inhibitor (Roche, #11836170001) was added to this lysis buffer.&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">Microscope</td>
			<td style="width:181px;">Zeiss</td>
			<td style="width:175px;">LSM510</td>
			<td style="width:499px;">Red, green and blue channels to visualize AF594, AF488 and DAPI fluorescent immunostainings (40X mag).</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Mounting media (Hydromount)</td>
			<td style="width:181px;">National Diagnostics</td>
			<td style="width:175px;">HS-106</td>
			<td>Fluorescent staining</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Mounting media (Cytoseal)</td>
			<td style="width:181px;">ThermoFisher Scientific</td>
			<td style="width:175px;">8310-16</td>
			<td>Histology (HRP)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">OCT compound (Tissue Tek)</td>
			<td style="width:181px;">Sakura</td>
			<td style="width:175px;">4583</td>
			<td>Embedding for cryotomy</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Penicillin-streptomycin antibiotics</td>
			<td style="width:181px;">Sigma Aldrich</td>
			<td style="width:175px;">A5955</td>
			<td>Cell culture media additive</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Primary antibodies</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Anti-Collagen I antibody</td>
			<td style="width:181px;">Abcam</td>
			<td style="width:175px;">#ab6308</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Anti-Collagen Type IV</td>
			<td style="width:181px;">Novocastra</td>
			<td style="width:175px;">#NCL-COLL-IV</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Anti-collagen VII</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>LH7.2 (in house)&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Anti-Fibronectin antibody</td>
			<td style="width:181px;">Abcam</td>
			<td style="width:175px;">#ab2413</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Reducing agent&#160;</td>
			<td style="width:181px;">ThermoFisher Scientific</td>
			<td style="width:175px;">NP0009</td>
			<td>Western blot</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Sample buffer</td>
			<td style="width:181px;">ThermoFisher Scientific</td>
			<td style="width:175px;">NP0008</td>
			<td>Western blot</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Secondary antibodies (Immunostaining)</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">4&#8242;,6-Diamidino-2-phenylindole dihydrochloride (DAPI)</td>
			<td style="width:181px;">Sigma Aldrich</td>
			<td style="width:175px;">#D9542</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">AlexaFluor 594 goat-anti-rabbit</td>
			<td style="width:181px;">ThermoFisher Scientific</td>
			<td style="width:175px;">#A-11037&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">AlexaFluor 594 goat-anti-mouse</td>
			<td style="width:181px;">ThermoFisher Scientific</td>
			<td style="width:175px;">&#160;#A-11005</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">AlexaFluor 488 chicken-anti-rabbit</td>
			<td style="width:181px;">ThermoFisher Scientific</td>
			<td style="width:175px;">#A-21441</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">AlexaFluor 488 goat-anti-mouse</td>
			<td style="width:181px;">ThermoFisher Scientific</td>
			<td style="width:175px;">#A-11001</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:299px;">Secondary antibodies (HRP)</td>
			<td style="width:181px;">Dako</td>
			<td style="width:175px;">Envision + System - HRP Labelled Polymer Anti-Rabbit and Anti-Mouse</td>
			<td>undiluted</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">Sodium deoxycholate</td>
			<td style="width:181px;">Prodotti Chimicie Alimentari</td>
			<td style="width:175px;"></td>
			<td>Decellularization</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Stratification media</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Dulbecco&#39;s Modified Eagle&#39;s Medium&#160;</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>Used during the air-liquid interface of organotypic cultures. This media is added to the outside of the cell-culture insert.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Ham&#8217;s F12&#160;&#160;&#160;</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>Used during the air-liquid interface of organotypic cultures. This media is added to the outside of the cell-culture insert.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">10% fetal bovine serum&#160;</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>Used during the air-liquid interface of organotypic cultures. This media is added to the outside of the cell-culture insert.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">100U/mL penicillin and 100U/mL streptomycin&#160;&#160;</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>Used during the air-liquid interface of organotypic cultures. This media is added to the outside of the cell-culture insert.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">0.4mg/mL hydrocortisone&#160;&#160;&#160;</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>Used during the air-liquid interface of organotypic cultures. This media is added to the outside of the cell-culture insert.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">5mg/mL insulin</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>Used during the air-liquid interface of organotypic cultures. This media is added to the outside of the cell-culture insert.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">1.8&#183;10-4 M adenine&#160;&#160;</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>Used during the air-liquid interface of organotypic cultures. This media is added to the outside of the cell-culture insert.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">5mg/mL transferrin&#160;</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>Used during the air-liquid interface of organotypic cultures. This media is added to the outside of the cell-culture insert.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">2&#183;10- 11 M triiodothyronine&#160;</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td>Used during the air-liquid interface of organotypic cultures. This media is added to the outside of the cell-culture insert.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">Trypsin</td>
			<td style="width:181px;">Biopolis Shared Facilities</td>
			<td style="width:175px;">0.125% Trypsin/Versene&#160;&#160; pH 7.0 + 0.3&#160;</td>
			<td>Used to trypsinize cells</td>
		</tr>
	</tbody>
</table>

improving 2d and 3d skin in vitro models using macromolecular crowding

Den glycoprotein familie af collagener repræsenterer de vigtigste strukturelle proteiner i den menneskelige krop, og er centrale elementer i biomaterialer, der anvendes i moderne tissue engineering. En teknisk flaskehals er aflejringen af collagen in vitro, som det er notorisk langsomt, hvilket resulterer i suboptimal dannelse af bindevæv og efterfølgende væv samhørighed, især i hudmodeller. Her beskriver vi en fremgangsmåde, der indebærer tilsætning af differentielt størrelse saccharose copolymerer i skindet kulturer til at generere makromolekylær fortrængning (MMC), hvilket resulterer i en dramatisk forøgelse af kollagen deposition. Især dermale fibroblaster deponeret en betydelig mængde af kollagen I / IV / VII og fibronectin under MMC sammenlignet med kontroller. Protokollen beskriver også en fremgangsmåde til decellularize overfyldte cellelag, udsætter betydelige mængder af ekstracellulær matrix (ECM), som blev tilbageholdt på kulturen overflade som vist ved immunocytochemistry. Total matrix masse og fordelingsmønster blev undersøgt under anvendelse interferens refleksioner mikroskopi. Interessant fibroblaster, keratinocytter og co-kulturer fremstillet celleafledte matricer (CDM) af varierende sammensætning og morfologi. CDM kunne bruges som &quot;bio-stilladser&quot; for sekundær celle såning, hvor den nuværende anvendelse af belægninger eller stilladser, typisk fra xenogene animalske kilder, kan undgås, og dermed bevæger sig i retning af mere klinisk relevant applikationer. Derudover denne protokol beskriver anvendelsen af ​​MMC i den neddykkede fase af en 3D-organotypiske hud co-kultur model, som var tilstrækkelig til at forøge ECM aflejring i dermo-epidermal junction (DEJ), især kollagen VII, hovedkomponenten forankringsorganer fibriller. Elektronmikroskopi bekræftede tilstedeværelsen af ​​forankring fibriller i kulturer udviklet med MMC, sammenlignet med kontroller. Dette er væsentligt, da forankringsorganer fibriller tether dermis til epidermis dermedhar en foruddannet modne DEJ kan drage fordel hudtransplantation modtagere i form af graft stabilitet og samlet sårheling. Endvidere blev dyrkningstid kondenseret fra 5 uger til 3 uger for at opnå en moden konstruktion, når der anvendes MMC, reducere omkostningerne.

Macromolecular fortrængning kunne forbedre ECM deposition, navnlig fibroblaster deponeret mere kollagen I, IV og fibronektin sammenlignet med kontrolkulturer (Figur 1, Cell lag; 1A, collagen I, 1B, collagen IV, 1C, fibronectin). Ved decellularization, det var tydeligt, at fibroblaster var de vigtigste indskydere kollagen I, IV og fibronektin i forhold til keratinocytter (figur 1, Matrix). <p class="...

Watch this Scientific Journal Video about Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding at JoVE.com

Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding

Vi præsenterer en protokol for at opnå celle-afledte matricer rige på ekstracellulære matrixproteiner, hjælp makromolekylære Crowders (MMC). Derudover præsenterer vi en protokol, som inkorporerer MMC i 3D organotypisk hud co-kultur generation, hvilket reducerer dyrkningstid samtidig opretholde løbetid på konstruktionen.

Forbedring 2D og 3D Skin<em&gt; In vitro</em&gt; Modeller Brug Macromolecular Fortrængning

The glycoprotein family of collagens represents the main structural proteins in the human body, and are key components of biomaterials used in modern ...

Research

JoVE Journal

Bioengineering

Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding

Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition

Visualisere Proteiner og makromolekylære Komplekser af negative Stain EM: fra Grid Forberedelse til Image Acquisition

Single particle electron microscopy (EM), of both negative stained or frozen hydrated biological samples, has become a versatile tool in structural ...

Electric Field-controlled Directed Migration of Neural Progenitor Cells in 2D and 3D Environments

Elektrisk felt-styret Instrueret Migration af neurale stamceller i 2D-og 3D-miljøer

Endogenous electric fields (EFs) occur naturally in vivo and play a critical role during tissue/organ development and regeneration, including that of the ...

Cell Co-culture Patterning Using Aqueous Two-phase Systems

Celle co-kultur Patterning hjælp Vandige tofasesystemer

Cell patterning  technologies that are fast, easy to use and affordable will be required for the  future development of high throughput cell assays, ...

Longitudinal Measurement of Extracellular Matrix Rigidity in 3D Tumor Models Using Particle-tracking Microrheology

Longitudinal Måling af ekstracellulær matrix Stivhed i 3D tumormodeller med Partikel-tracking Microrheology

The mechanical microenvironment has been shown to act as a crucial regulator of tumor growth behavior and signaling, which is itself remodeled and ...

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

Øjenirritation Test (EIT) for Hazard Identifikation af Eye Irriterer Kemikalier hjælp rekonstrueret human Cornea-lignende Epithelial (RHCE) Tissue Model

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

2D and 3D Matrices to Study Linear Invadosome Formation and Activity

2D og 3D matricer til at studere lineær invadosomdannelse og aktivitet

Cell adhesion, migration, and invasion are involved in many physiological and pathological processes. For example, during metastasis formation, tumor ...

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

Bioprinting af brusk og huden væv analoger udnytter en roman passiv blanding enhed teknik for Bioink Precellularization

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

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

Forbedret 3D Hydrogel kulturer af primære Glial celler for In Vitro modellering af Neuroinflammation

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

In Vitro Model of Human Cutaneous Hypertrophic Scarring using Macromolecular Crowding

In Vitro Model af Human Kutan hypertrofisk ardannelse ved hjælp af makromolekylære fortrængning

It has been shown that in vivo tissues are highly crowded by proteins, nucleic acids, ribonucleoproteins, polysaccharides, etc. The following protocol ...

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

Avancerede 3D-levermodeller til in vitro genotoksicitetstest efter langvarig nanomaterialeeksponering

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