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

Gli autori desiderano ringraziare Kent State University (assegno di ricerca in collaborazione e il supporto per la Medicina Rigenerativa iniziativa alla Kent State - ReMedIKS) per il sostegno finanziario di questo progetto.

NOTA: Le seguenti operazioni per la preparazione 3D LCE gomma-come utilizzando il blocco stella 3-braccio copolimero sono mostrati in Figura 1. Per la caratterizzazione Risonanza Magnetica Nucleare (NMR), gli spettri vengono registrati in cloroformio deuterato (CDCl 3) a temperatura su uno strumento Bruker DMX 400 MHz camera e internamente riferimento picchi residui a 7.26. Fourier Transform Infrared spettri (FT-IR) vengono registrati con un Bruker Vector 33 FT-IR spettrometro u...

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elastomeri liquido cristallini sono stati recentemente studiati come scaffold cellulari biocompatibili a causa della loro reattività stimoli. Essi hanno dimostrato di essere le piattaforme ideali come ponteggi cellulari. Tuttavia, un fattore importante da tenere a mente durante la preparazione e la progettazione di un nuovo scaffold LCE è la porosità. L&#39;incorporazione di solidi lisciviabili 23 o gas non sempre comporta porosità omogenea o pori completamente interconnessi. L&#39;uso di un ...

Esistono numerosi esempi di materiali sintetici biologici e biocompatibili destinati ad essere applicati in studi cellulari e per la rigenerazione tissutale obiettivo di adesione cellulare e la proliferazione 1, 2, 3, 4, 5. Ci sono stati alcuni esempi di materiali biocompatibili, noti come elastomeri a cristalli liquidi (LCEs), che potrebbe rispondere a stimoli esterni con anisotropico molecolare ordinare 6, 7. LCEs sono materiali di stimoli reattivi che combinano le proprietà meccaniche ed elastiche elastomeri con la funzionalità ottica e ordine molecolare di cristalli liquidi 8, 9. LCEs possono sperimentare cambiamenti di forma, deformazione meccanica, comportamento elastico, e proprietà ottiche in risposta a STIM esterniuli (es., calore, stress, luce, ecc) 10, 11, 12, 13, 14, 15, 16. Studi precedenti hanno dimostrato che i cristalli liquidi (LC) possono percepire la crescita e l&#39;orientamento delle cellule 4, 17. È quindi possibile supporre che LCEs possono essere adatti per applicazioni biologicamente e clinicamente rilevante, compresi ponteggi cellulare e allineamento. Abbiamo già riferito la preparazione di smectiche film biocompatibili, biodegradabili, cast-stampati, e LCEs sottili con un &quot;tipo Swiss-formaggio&quot; morfologia porosa 6, 18. Abbiamo anche preparato LCEs biocompatibili nematici con morfologia sferica come supporti per la crescita cellulare 19 &lt;sup&gt;, 20. Il nostro lavoro è stato rivolto a punto delle proprietà meccaniche dei materiali da corrispondere a quelli del tessuto di interesse 21. Inoltre, questi studi si concentrano sulla comprensione interazioni elastomero-cellula, così come risposta cellulare quando gli elastomeri sono soggetti a stimoli esterni. Le sfide principali erano in parte per adattare la porosità dei LCEs per permettere l&#39;attacco cellulare e permeazione attraverso la matrice elastomerica e per un migliore trasporto di massa. La porosità di questi film sottili 6 consentito per permeazione attraverso la massa delle cellule della matrice, ma non tutti i pori erano completamente interconnesso o aveva una dimensione più regolare (omogenea) poro. Abbiamo poi riportato su biocompatibili elastomeri nematici LCE con morfologie globulari. Questi elastomeri nematici consentiti per il fissaggio e la proliferazione delle cellule, ma la dimensione dei pori compresa soltanto 10-30 um, che ha impedito o limitato l&#39;uso di questielastomeri con una più ampia varietà di linee cellulari 19, 20. Precedente lavoro di Kung et al. relativa alla formazione di schiume grafene utilizzando un modello metal &quot;sacrificale&quot; si evince che la schiuma grafene ottenuto aveva una morfologia porosa molto regolare dettata dal modello metallo scelto 22. Questa metodologia offre il pieno controllo di porosità e dimensione dei pori. Allo stesso tempo, la malleabilità e flessibilità del modello metallo consentono la formazione di diverse forme template prima della preparazione di schiuma. Altre tecniche, come lisciviazione materiale 23, templating gas 24, o fibre elettro-filato 25, 26 offrono anche il potenziale per la preparazione di materiali porosi, ma sono più tempo e, in alcuni casi, la dimensione dei pori è limitata a solo pochi micrometri. Schiuma-come LCEs 3D preparati utilizzando forme in metallo consentono una cella di carico più elevato; un tasso di proliferazione migliorata; co-coltura; e, non ultimo, una migliore gestione del trasporto di massa (cioè, nutrienti, gas e rifiuti) per assicurare sviluppo tessuto pieno 27. LCEs 3D gomma-come appaiono anche per migliorare l&#39;allineamento delle cellule; questo è più probabile in relazione alle pendenti LC sensing crescita cellulare e l&#39;orientamento delle cellule. La presenza di frazioni LC all&#39;interno LCE sembra aumentare allineamento delle celle rispetto alla posizione della cella all&#39;interno del scaffold LCE. Cellule allineati entro i puntoni della LCE, mentre nessun chiaro orientamento si osserva in cui i puntoni uniscono (giunzioni) 27. Nel complesso, la nostra piattaforma impalcatura cella LCE fungere da supporto cellulare offre la possibilità di regolare la morfologia elastomero e proprietà elastiche e di indirizzare specificamente l&#39;allineamento (individuali) tipi cellulari per creare un ordine, disposizioni spaziali ocellule F simili ai sistemi viventi. Oltre ad un&#39;impalcatura in grado di sostenere e dirigere la crescita cellulare a lungo termine e la proliferazione, LCEs anche consentire esperimenti dinamici, in cui l&#39;orientamento delle cellule e interazioni possono essere modificate al volo.

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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>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-propanol&#160;</td>
			<td>Sigma Aldrich</td>
			<td>278475</td>
			<td>Solvent</td>
		</tr>
		<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>
		</tr>
		<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>
		<tr height="21">
			<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

Qui vi presentiamo una preparazione step-by-step di un 3D, biodegradabili, gomma-come impalcatura delle cellule. Tali ponteggi sono stati preparati mediante reticolazione blocchi a stella copolimeri dotati di unità di colesterolo come gruppi pendenti catena laterale, con conseguente smectic-A (SMA) elastomeri a cristalli liquidi (LCEs). impalcature Schiuma-like, preparati utilizzando forme in metallo, sono dotate di microcanali interconnessi, che li rende adatti come impalcature di coltura cellulare 3D. Le proprietà combinate della struttura regolare della schiuma metallo e del risultato elastomero in un&#39;impalcatura cellule 3D che favorisce non solo superiore proliferazione cellulare rispetto alle tradizionali pellicole templated porosi, ma anche una migliore gestione del trasporto di massa (cioè, nutrienti, gas, rifiuti , ecc). La natura del modello di metallo permette una facile manipolazione del materiale espanso (cioè, rotoli o film) e per la preparazione di supporti di differenti dimensioni dei pori per diversi studi sulle cellule preservando di interfacciamentoted natura porosa del modello. Il processo di incisione non influenza la chimica degli elastomeri, conservando la loro natura biocompatibile e biodegradabile. Abbiamo dimostrato che questi LCEs smectiche, quando coltivato per lunghi periodi di tempo, permettono lo studio di clinicamente rilevanti e complessi costrutti di tessuto, favorendo la crescita e la proliferazione delle cellule.

Questo rapporto mostra il metodo di preparazione di un LCE 3D poroso come impalcatura per la coltura cellulare utilizzando un modello di nichel metallico. Il ottenuto 3D LCE dimostra una complessa rete di canali interconnessi che permette una facile infiltrazione di cellule, così come più adatto trasporto di massa 27. Si è constatato che le cellule sono in grado di penetrare completamente la rete di canali interconnessi e sono anche in grado di allineare all&#3...

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

Questo studio presenta una metodologia per preparare 3D, biodegradabili, schiuma come scaffold cellulari basate su elastomeri cristalli liquidi catena laterale biocompatibili (LCEs). esperimenti di microscopia confocale mostrano che LCEs schiuma come permettere l&#39;attacco, proliferazione, e l&#39;allineamento spontaneo dei mioblasti C2C12s.

Sintesi di biocompatibili a cristalli liquidi in elastomero Schiume come impalcature cellulari per 3D spaziali colture cellulari

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

Microcapsule di alginato come piattaforma 3D per la propagazione e la differenziazione delle cellule staminali embrionali umane (hESC) per diverse linee

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

Ricerca

Bioingegneria

Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications

Preparazione del 3D fibrina ponteggi per le applicazioni delle cellule staminali Cultura

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

Ponteggi self-reporting per coltura cellulare 3-Dimensional

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

Fabbricazione di meccanicamente Tunable e Bioactive metallo Ponteggi per applicazioni biomediche

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

Alternando microgel ibrida Gelatina di campo magnetico-Responsive per Controlled Drug uscita

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

Preparazione di Monodomain Liquid Crystal Elastomeri e cristallo elastomero Nanocompositi Liquid

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 e montaggio dei microvasi 3D

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

Fabbricazione di Schiume matrice extracellulare-derivati ​​e microcarriers come tessuto-specifica delle cellule, Cultura e piattaforme di distribuzione

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

Un microcontrollore azionato il dispositivo per la generazione di estratti liquidi da Aerosol elettronico e fumo di sigaretta sigaretta convenzionale

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

Analisi 3D delle risposte multi-cellulari ai gradienti chemoattractant

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