Name: synthesis of biocompatible liquid crystal elastomer foams as cell scaffolds for 3d spatial cell cultures
Uploaded: 2017-04-11T15:00:00
Description: Watch this Scientific Journal Video about Synthesis of Biocompatible Liquid Crystal Elastomer Foams as Cell Scaffolds for 3D Spatial Cell Cultures at JoVE.com

Forfatterne ønsker å takke Kent State University (forskningssamarbeid stipend og støtte for Regenerative Medicine Initiative ved Kent State - ReMedIKS) for økonomisk støtte fra dette prosjektet.

MERK: Følgende trinn for 3D-LCE skumlignende preparat ved hjelp av tre-armet stjerne-blokk-kopolymer er vist i figur 1. For kjernemagnetisk resonans (NMR) karakterisering, er spektrene tatt opp i deuterert kloroform (CDCI3) ved romtemperatur på et Bruker DMX 400 MHz instrument og internt referert rest topper ved 7,26. Fourier transform infrarød (FT-IR) spektra ble registrert med en Bruker Vector 33 FT-IR spektrometer ved anvendelse av attenuert total reflektans. For hvert tri...

<ol>
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Flytende krystallinske elastomerer har nylig blitt studert som biokompatible cellestillasene på grunn av deres reaksjonsevne stimuli. De har vist seg å være ideelle plattformer som celle stillaser. Men en viktig faktor å huske på når du forbereder og utforme en ny LCE stillaset er porøsitet. Inkorporeringen av utlutbare faststoffer 23 eller gasser ikke alltid resulterer i homogen porøsitet eller fullstendig innbyrdes forbundet porer. Bruken av et metall mal som kan være etset seg ikke ba...

Det er flere eksempler på biologiske og biokompatible syntetiske materialer beregnet for anvendelse i cellestudier og for vevsregenerering sikte på celleadhesjon og celleformering 1, 2, 3, 4, 5. Det har vært noen få eksempler på biokompatible materialer, kjent som flytende krystall elastomerer (LCEs), som kan reagere på ytre stimuli med anisotropisk molekyl bestilling 6, 7. LCEs er stimuli-responsive materialer som kombinerer mekaniske og elastiske egenskaper for elastomerer med optisk funksjonalitet og molekyl ordning av flytende krystaller 8, 9. LCEs kan oppleve endringer i form, mekanisk deformasjon, elastisk oppførsel, og optiske egenskaper som respons på ytre stimuli (f.eks., varme, stress, lys, etc.) 10, 11, 12, 13, 14, 15, 16. Tidligere studier har vist at flytende krystaller (LCS) kan ane vekst og orientering av celler 4, 17. Det er mulig deretter å anta at LCEs kan være egnet for biologisk og medisinsk relevant applikasjoner, inkludert celle stillas og innretting. Vi har tidligere rapportert fremstillingen av smektiske biokompatible, bionedbrytbare, cast-formete, og tynne LCEs filmer med en &quot;Swiss-ostetype&quot; porøs morfologi 6, 18. Vi har også fremstilt nematiske biokompatible LCEs med kuleformede morfologi som stillas for cellevekst 19 &lt;sup&gt; 20. Vårt arbeid var rettet mot tuning de mekaniske egenskapene til materialer for å matche de av vevet av interesse 21. Også disse studier fokuserer på å forstå elastomer-celleinteraksjoner, så vel som den cellulære reaksjonen når elastomerene er utsatt for eksterne stimuli. De største utfordringene ble delvis å skreddersy porøsiteten til LCEs for å gi rom for cellebinding og gjennomtrengning gjennom det elastomere matrise og for bedre massetransport. Porøsiteten av disse tynne filmer 6 er tillatt for cellegjennomtrengning gjennom hoveddelen av matrisen, men ikke alle porer var fullstendig innbyrdes forbundet, eller hadde en mer vanlig (homogen) er porestørrelsen. Vi rapporterte på biokompatible nematiske LCE elastomerer med kule morfologi. Disse nematisk elastomerer er tillatt for festing og formering av celler, men porestørrelsen varierte bare 10-30 um, som hindret eller begrenset bruken av disseelastomerer med et bredere utvalg av cellelinjer 19, 20. Tidligere arbeid av Kung et al. i forbindelse med dannelsen av graphene skum ved anvendelse av en &quot;offer&quot; metall mal viste at det oppnådde graphene Skummet hadde en meget vanlig porøs morfologi bestemt av det valgte metall malen 22. Denne metodikken gir full kontroll av porøsitet og pore størrelse. På samme tid, formbarhet og fleksibilitet av metallet malen muliggjøre dannelse av annen mal former før skumfremstilling. Andre teknikker, slik som materiale utvasking 23, gass sjablonmiddel 24, eller elektro-spunnet fiber 25, 26 har også potensialet for fremstilling av porøse materialer, men de er mer tidkrevende og, i noen tilfeller, er porestørrelsen begrenset til bare noen få mikrometer. skum-lignende 3D LCEs fremstilt ved anvendelse av metall-maler tillate en høyere celle belastning; en forbedret spredning hastighet; co-dyrkning; og, sist men ikke minst, bedre massetransport management (dvs. næringsstoffer, gasser og avfall) sikre full vev utvikling 27. Skum-lignende 3D LCEs synes også å forbedre celle justering; Dette er mest sannsynlig i forhold til de LC anheng avføler cellevekst og celle orientering. Tilstedeværelsen av LC grupper innenfor LCE ser ut til å forbedre celle innretting med hensyn til celle sted inne i LCE stillaset. Celler justeres innenfor avstiverne av LCE, mens ingen klar orientering observeres hvor stiverne går sammen (kryss) 27. Samlet vår LCE celle stillasgulv som en celle-støttemedium gir muligheter for å justere elastomer morfologi og elastiske egenskaper og for spesifikt å styre innrettingen av (enkelt) celletyper for å skape en ordnet, romlige arrangementer of celler som ligner på levende systemer. Bortsett fra å tilveiebringe et stillas i stand til å støtte og lede langvarig cellevekst og proliferasjon, LCEs også tillate dynamiske eksperimenter, hvor cellen orientering og interaksjoner kan modifiseres på fly.

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			<td height="21" style="height:21px;width:291px;">1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane</td>
			<td style="width:148px;">Alfa Aesar</td>
			<td style="width:132px;">L16606</td>
			<td style="width:153px;">Silanizing agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-bis(4-hydroxy-cyclohexyl)propane</td>
			<td>TCI</td>
			<td><a href="http://www.tcichemicals.com/eshop/en/us/commodity/B0928/">B0928</a></td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-chlorohexanone&#160;</td>
			<td>Alfa Aesar</td>
			<td>A18613</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-heptanone&#160;</td>
			<td>Sigma Aldrich</td>
			<td>W254401</td>
			<td>Solvent</td>
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		<tr height="21">
			<td height="21" style="height:21px;">2-propanol&#160;</td>
			<td>Sigma Aldrich</td>
			<td>278475</td>
			<td>Solvent</td>
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		<tr height="21">
			<td height="21" style="height:21px;">3-chloroperbenzoic acid, m-CPBA</td>
			<td>Sigma Aldrich</td>
			<td>273031</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4-dimethylaminopyridine</td>
			<td>Alfa Aesar</td>
			<td>A13016</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4&#39;,6-diamidino-2-phenylindole, DAPI&#160;</td>
			<td>Invitrogen</td>
			<td>D1306</td>
			<td>Nuclear Stain</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5-hexynoic acid&#160;</td>
			<td>Alfa Aesar</td>
			<td>B25132-06</td>
			<td>Reagent</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Acetic acid</td>
			<td>VWR</td>
			<td>36289</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Acetone</td>
			<td>Sigma Aldrich</td>
			<td>34850</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alcohol 200 proof ACS Grade&#160;</td>
			<td>VWR</td>
			<td>71001-866</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Benzene</td>
			<td>Alfa Aesar</td>
			<td>AA33290</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#949;-caprolactone&#160;</td>
			<td>Alfa Aesar</td>
			<td>A10299-0E</td>
			<td>Reagent</td>
		</tr>
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			<td height="21" style="height:21px;">Chloroform</td>
			<td>VWR</td>
			<td>BDH1109</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cholesterol</td>
			<td>Sigma Aldrich</td>
			<td>C8503</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Chromium(VI) oxide</td>
			<td>Sigma Aldrich</td>
			<td>232653</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Copper (I) iodide</td>
			<td>Strem Chemicals</td>
			<td>100211-060</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">D,L-Lactide&#160;</td>
			<td>Alfa Aesar</td>
			<td>L09026</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dichloromethane</td>
			<td>Sigma Aldrich</td>
			<td>320269</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Diethyl ether&#160;</td>
			<td>Emd Millipore</td>
			<td>EX0190</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">N,N-Dimethylformamide</td>
			<td>Sigma Aldrich</td>
			<td>270547</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dulbecco&#8217;s modified Eagle medium, DEME&#160;</td>
			<td>CORNING Cellgo</td>
			<td>10-013</td>
			<td>Cell Media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethanol</td>
			<td>Alfa Aesar</td>
			<td>33361</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Formaldehyde&#160;</td>
			<td>SIGMA Life Science</td>
			<td>F8775</td>
			<td>Fixative</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fetal bovine serum, FBS&#160;</td>
			<td>HyClone</td>
			<td>SH30071.01</td>
			<td>Media Component</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Filter paper, Grade 415, qualitative, crepe</td>
			<td>VWR</td>
			<td>28320</td>
			<td>Filtration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glycerol</td>
			<td>Sigma Aldrich</td>
			<td>G5516</td>
			<td>Central node (3-arm)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hexamethylene diisocyanate, HDI</td>
			<td>Sigma Aldrich</td>
			<td>52649</td>
			<td>Crosslinker</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Iron(III) chloride&#160;</td>
			<td>Alfa Aesar</td>
			<td>12357</td>
			<td>Etching agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isopropyl alcohol</td>
			<td>VWR</td>
			<td>BDH1133</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Methanol</td>
			<td>Alfa Aesar</td>
			<td>L13255</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">N,N&#39;-dicyclohexylcarbodiimide</td>
			<td>Aldrich</td>
			<td>D80002</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">N,N-Dimethylformamide</td>
			<td>Sigma Aldrich</td>
			<td>270547</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nickel metal template</td>
			<td>American Elements</td>
			<td>Ni-860</td>
			<td>Foam template</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Neuroblastomas cells (SH-SY5Y)</td>
			<td>ATCC</td>
			<td>CRL-2266</td>
			<td>Cell line</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Penicillin streptomycin&#160;</td>
			<td>Thermo SCIENTIFIC</td>
			<td>15140122</td>
			<td>Antibiotics</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polyethylene glycol 2000, PEG</td>
			<td>Alfa Aesar</td>
			<td>B22181</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium azide&#160;</td>
			<td>VWR</td>
			<td>97064-646</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium bicarbonate</td>
			<td>AMRESCO</td>
			<td>865</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium chloride</td>
			<td>BDH</td>
			<td>BDH9286</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium phosphate dibasic heptahydrate</td>
			<td>Fisher Scientific</td>
			<td>S-374</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium phosphate monobasic monohydrate</td>
			<td>Sigma Aldrich</td>
			<td>S9638</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium sulfate</td>
			<td>Sigma Aldrich</td>
			<td>239313</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tetrahydrofuran</td>
			<td>Alfa Aesar</td>
			<td>41819</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Thiosulfate de sodium</td>
			<td>AMRESCO</td>
			<td>393</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tin(II) 2-ethylhexanoate</td>
			<td>Aldrich</td>
			<td>S3252</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Toluene&#160;</td>
			<td>Alfa Aesar</td>
			<td>22903</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triethylamine</td>
			<td>Sigma Aldrich</td>
			<td>471283</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Trypsin&#160;</td>
			<td>HyClone</td>
			<td>SH30042.01</td>
			<td>Cell Detachment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Olympus FV1000</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

synthesis of biocompatible liquid crystal elastomer foams as cell scaffolds for 3d spatial cell cultures

Her presenterer vi en trinn-for-trinn fremstillingen av en 3D, bionedbrytbar, skumlignende celle stillaset. Disse stillaser ble fremstilt ved tverrbinding av stjerne-blokk-kopolymerer Med kolesterolheter som sidekjede utstikkende grupper, noe som resulterer i smektisk A-(SMA) med flytende krystall-elastomer (LCEs). Skumlignende stillaser, fremstilt ved anvendelse av metallrammer, har innbyrdes forbundne mikrokanaler, noe som gjør dem egnet som 3D-cellekultur stillaser. De kombinerte egenskapene til den vanlige struktur av metallskum og av elastomeren resultat i et 3D-celle stillas som fremmer ikke bare høyere celleproliferasjon sammenlignet med konvensjonelle porøse malbasert filmer, men også bedre styring av massetransport (dvs. næringsstoffer, gasser, avfalls , etc.). Naturen av metall malen gir en enkel manipulering av skum former (dvs. ruller eller filmer) og for fremstilling av stillaser med forskjellige porestørrelser for forskjellige cellestudier og samtidig bevare interconnected porøse natur av malen. Etseprosessen har ingen innvirkning på kjemien av elastomerene, bevare deres biokompatible og biologisk nedbrytbare natur. Vi viser at disse smektiske LCEs, når dyrket for omfattende tidsperioder, aktiverer studier av klinisk relevante og komplekse vev konstruerer mens fremme vekst og spredning av celler.

Denne rapporten viser fremgangsmåten for fremstilling av en porøs 3D LCE som et stillas for cellekultur ved anvendelse av en nikkel-metall-mal. Den oppnådde 3D LCE viser et komplekst sammenkoplet kanal nettverk som gir mulighet for enkel celleinfiltrasjon, så vel som mer egnet massetransporten 27. Det ble funnet at celler som er i stand til fullt ut trenge inn i det sammenkoplede kanalnettet og er også i stand til å justere innenfor LCE. Her, et metall nikke...

Watch this Scientific Journal Video about Synthesis of Biocompatible Liquid Crystal Elastomer Foams as Cell Scaffolds for 3D Spatial Cell Cultures at JoVE.com

Synthesis of Biocompatible Liquid Crystal Elastomer Foams as Cell Scaffolds for 3D Spatial Cell Cultures

Denne studien viser en metode for å fremstille 3D, biologisk nedbrytbare, skumlignende cellestillasene basert på biokompatible sidekjede flytende krystall-elastomerer (LCEs). Konfokalmikroskopi forsøk viser at skumlignende LCEs at cellene skulle feste, proliferasjon, og den spontane innretting av C2C12s myoblaster.

Syntese av Biokompatible Liquid Crystal Elastomer Skum som Cell Stillas for 3D Spatial cellekulturer

Here, we present a step-by-step preparation of a 3D, biodegradable, foam-like cell scaffold. These scaffolds were prepared by cross-linking star block ...

Research

JoVE Journal

Bioengineering

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells (hESC) to Different Lineages

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages

Alginat Microcapsule som en 3D-plattform for Formering og Differensiering av humane embryonale stamceller (hESC) til ulike linjene

Human embryonic stem cells (hESC) are emerging as an attractive alternative source for cell replacement therapy since they can be expanded in culture ...

Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications

Utarbeidelse av 3D Fibrin stillaser for Stem Cell Kultur Applications

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells ...

Self-reporting Scaffolds for 3-Dimensional Cell Culture

Egenrapportering Stillaser for 3-dimensjonal Cell Culture

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting ...

Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

Fabrikasjon av mekanisk Tunable og Bioaktive Metal Stillas for Biomedical Applications

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness ...

Alternating Magnetic Field-Responsive Hybrid Gelatin Microgels for Controlled Drug Release

Vekslende magnetfelt-Responsive Hybrid Gelatin mikrogeler for kontrollert legemiddeldosering

Magnetically-responsive nano/micro-engineered biomaterials that enable a tightly controlled, on-demand drug delivery have been developed as new types of ...

Preparation of Monodomain Liquid Crystal Elastomers and Liquid Crystal Elastomer Nanocomposites

Utarbeidelse av monodomene flytende krystall elastomerer og flytende krystall Elastomer Nanocomposites

LCEs are shape-responsive materials with fully reversible shape change and potential applications in medicine, tissue engineering, artificial muscles, and ...

Micropatterning and Assembly of 3D Microvessels

Micropatterning og montering av 3D microvessels

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

Fabrication of Extracellular Matrix-derived Foams and Microcarriers as Tissue-specific Cell Culture and Delivery Platforms

Fabrikasjon av ekstracellulære matrise-avledet Skum og Mikrocarriers som Tissue-spesifikke cellekultur og Delivery plattformer

Cell function is mediated by interactions with the extracellular matrix (ECM), which has complex tissue-specific composition and architecture. The focus ...

A Microcontroller Operated Device for the Generation of Liquid Extracts from Conventional Cigarette Smoke and Electronic Cigarette Aerosol

En Microcontroller drives enhet for generering av flytende ekstrakter fra konvensjonelle sigarett røyk og elektronisk sigarett Aerosol

Electronic cigarettes are the most popular tobacco product among middle and high schoolers and are the most popular alternative tobacco product among ...

3D Analysis of Multi-cellular Responses to Chemoattractant Gradients

3D-analyse av multi-Cellular Svar å Chemoattractant graderinger

Various limitations of 2D cell culture systems have sparked interest in 3D cell culture and analysis platforms, which would better mimic the spatial and ...