Name: constructing a collagen hydrogel for the delivery of stem cell-loaded chitosan microspheres
Uploaded: 2012-06-01T12:00:00
Description: Watch this Scientific Journal Video about Constructing a Collagen Hydrogel for the Delivery of Stem Cell-loaded Chitosan Microspheres at JoVE.com

DOZ understøttes af et tilskud fra Genève Foundation. SN blev understøttet af en postdoc Fellowship Grant fra Pittsburgh Tissue Engineering Initiative.

1. Isolerende adipøst afledte stamceller (ASC) Bemærk: Alle procedurer blev udført ved stuetemperatur, medmindre andet er angivet. <ol><li> Isoler rotte perirenalt og epididymalt fedt-og vaskes med steril Hanks pufrede saltopløsning (HBSS) indeholdende 1% føtalt bovint serum (FBS) som tidligere beskrevet 6. </li><li> Hakket vævet og overføre 1-2 g i 25 ml HBSS indeholdende 1% FBS i et 50 ml rør og centrifugeres ved 500 g i 8 minutter ved stuetemperatur. ...

<ol>
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(Praha). 57, 47-56 (2011).</li><li>Baruch, L., Machluf, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alginate-chitosan+complex+coacervation+for+cell+encapsulation:+effect+on+mechanical+properties+and+on+long-term+viability.">Alginate-chitosan complex coacervation for cell encapsulation: effect on mechanical properties and on long-term viability.</a> Biopolymers. 82, 570-579 (2006).</li><li>Wei, Y., Gong, K., Zheng, Z., Wang, A., Ao, Q., Gong, Y., Zhang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chitosan/silk+fibroin-based+tissue-engineered+graft+seeded+with+adipose-derived+stem+cells+enhances+nerve+regeneration+in+a+rat+model.">Chitosan/silk fibroin-based tissue-engineered graft seeded with adipose-derived stem cells enhances nerve regeneration in a rat model.</a> J. Mater. Sci. Mater. Med. , (2011).</li><li>Wang, Q., Jamal, S., Detamore, M. S., Berkland, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PLGA-chitosan/PLGA-alginate+nanoparticle+blends+as+biodegradable+colloidal+gels+for+seeding+human+umbilical+cord+mesenchymal+stem+cells.">PLGA-chitosan/PLGA-alginate nanoparticle blends as biodegradable colloidal gels for seeding human umbilical cord mesenchymal stem cells.</a> J. Biomed. Mater. Res. A. 96, 520-527 (2011).</li><li>Alves da Silva, M. L., Martins, A., Costa-Pinto, A. R., Correlo, V. M., Sol, P., Bhattacharya, M., Faria, S., Reis, R. L., Neves, N. 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N., Alt, E. U., Mathur, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adhesion,+migration+and+mechanics+of+human+adipose-tissue-derived+stem+cells+on+silk+fibroin-chitosan+matrix.">Adhesion, migration and mechanics of human adipose-tissue-derived stem cells on silk fibroin-chitosan matrix.</a> Acta Biomater. 6, 1388-1397 (2010).</li><li>Altman, A. M., Yan, Y., Matthias, N., Bai, X., Rios, C., Mathur, A. B., Song, Y. H., Alt, E. 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En stor hurdle i stamcelle-baseret terapi er ved at udvikle effektive metoder til levering af celler til de angivne områder til reparation. På grund af patient til patient variabilitet, den vævstype, tilskadekomst størrelse og dybde, skal metode til levering af stamceller afgøres fra sag til sag. Selv indlejring stamceller i en matrix og leverer dem til sårstedet synes at være en næste logiske fremgangsmåde til vævsmanipulering, nogle tekniske forhindringer tilbage. Dette omfatter muligheden af de indlejrede c...

<table border="1" ><tbody><tr><td> Navn på reagenset / udstyr </td><td> Firma </td><td> Katalog nummer </td><td> Kommentarer </td></tr><tr><td> Hanks BalancedSalt Solution (HBSS) </td><td> Gibco </td><td> 14175 </td><td> Varen </td></tr><tr><td> Føtalt bovint serum </td><td> Hyclone </td><td> SH30071.03 </td><td> Varen </td></tr><tr><td> Collagenase Type II </td><td> Sigma-Aldrich </td><td> C6685 </td><td> Varen </td></tr><tr><td> 70 um nylon mesh filter </td><td> BD Biosciences </td><td> 352350 </td><td> Varen </td></tr><tr><td> 100 um nylon mesh filter </td><td> BD Biosciences </td><td> 352360 </td><td> Varen </td></tr><tr><td> MesenPRO vækstmedium System </td><td> Invitrogen </td><td> 12746-012 </td><td> Varen </td></tr><tr><td> L-glutamin </td><td> Gibco </td><td> 25030 </td><td> Varen </td></tr><tr><td> T75 vævsdyrkningskolbe </td><td> BD Biosciences </td><td> 137787 </td><td> Varen </td></tr><tr><td> Chitosan </td><td> Sigma-Aldrich </td><td> 448869 </td><td> Varen </td></tr><tr><td> Eddikesyre </td><td> Sigma-Aldrich </td><td> 320099 </td><td> Varen </td></tr><tr><td> N-octanol </td><td> Acros Organics </td><td> 150630025 </td><td> Varen </td></tr><tr><td> Sorbitan-mono-oleat </td><td> Sigma-Aldrich </td><td> S6760 </td><td> Varen </td></tr><tr><td> Kaliumhydroxid </td><td> Sigma-Aldrich </td><td> P1767 </td><td> Varen </td></tr><tr><td> Acetone </td><td> Fisher Scientific </td><td> L-4859 </td><td> Varen </td></tr><tr><td> Ethanol </td><td> Sigma-Aldrich </td><td> 270741 </td><td> Varen </td></tr><tr><td> Trinitro benzensulfonsyre </td><td> Sigma-Aldrich </td><td> P2297 </td><td> Varen </td></tr><tr><td> Saltsyre </td><td> Sigma-Aldrich </td><td> 320331 </td><td> Varen </td></tr><tr><td> Ethylether </td><td> Sigma-Aldrich </td><td> 472-484 </td><td> Varen </td></tr><tr><td> 8-um vævskulturplade Inserts </td><td> BD Biosciences </td><td> 353097 </td><td> Varen </td></tr><tr><td> 1,5 ml-mikrocentrifugerør </td><td> Fisher </td><td> 05-408-129 </td><td> Varen </td></tr><tr><td> MTT Reagens </td><td> Invitrogen </td><td> M6494 </td><td> Varen </td></tr><tr><td> Dimethylsulfoxid </td><td> Sigma-Aldrich </td><td> D8779 </td><td> Varen </td></tr><tr><td> Qtracker Cell Labeling Kit (Q tracker 655) </td><td> Molekylære prober </td><td> Q2502PMP </td><td> Varen </td></tr><tr><td> Type 1 Collagen </td><td> Travigen </td><td> 3447-020-01 </td><td> Varen </td></tr><tr><td> Natriumhydroxid </td><td> Sigma-Aldrich </td><td> S8045 </td><td> Varen </td></tr><tr><td> 12-brønds vævskulturplader </td><td> BD Biosciences </td><td> 353043 </td><td> Varen </td></tr><tr><td> Centrifuger </td><td> Eppendorf </td><td> 5417R </td><td> Udstyr </td></tr><tr><td> Orbital Shaker </td><td> New Brunswick Scienctific </td><td> C24 </td><td> Udstyr </td&gt; </tr><tr><td> Befugtet inkubator med luft-5% CO2 </td><td> Thermo Scientific </td><td> Model 370 </td><td> Udstyr </td></tr><tr><td> Omrører </td><td> IKA </td><td> Visc6000 </td><td> Udstyr </td></tr><tr><td> Magnetomrører </td><td> Corning </td><td> PC-210 </td><td> Udstyr </td></tr><tr><td> Vakuumeksikator </td><td> - </td><td> - </td><td> Udstyr </td></tr><tr><td> Particle Size Analyzer </td><td> Malvern </td><td> STP2000 Spraytec </td><td> Udstyr </td></tr><tr><td> Vandbad </td><td> Fisher Scientific </td><td> Isotemp210 </td><td> Udstyr </td&gt; </tr><tr><td> Spektrofotometer </td><td> Beckman </td><td> Beckman Coulter DU800UV/Visible Spektrofotometer </td><td> Udstyr </td></tr><tr><td> Vortex </td><td> Diagger </td><td> 3030a </td><td> Udstyr </td></tr><tr><td> Mikropladelæser </td><td> Molecular Devices </td><td> SpectraMax M2 </td><td> Udstyr </td></tr><tr><td> Lys / fluorescensmikroskop </td><td> Olympus </td><td> IX71 </td><td> Udstyr </td></tr><tr><td> Konfokalt mikroskop </td><td> Olympus </td><td> FV-500 laser konfokal mikroskop </td><td> Udstyr </td></tr><tr><td> Scanning Electron Microscope </td><td> Carl Zeiss MicroImaging </td><td> Leo 435 VP </td><td> Udstyr </td></tr><tr><td> Transmission Electron Microscope </td><td> JEOL </td><td> JEOL 1230 </td><td> Udstyr </td></tr></tbody></table>

constructing a collagen hydrogel for the delivery of stem cell-loaded chitosan microspheres

Multipotente stamceller har vist sig at være yderst nyttig i forbindelse med regenerativ medicin 1-3. Imidlertid, for at bruge disse celler effektivt til vævsregenerering, skal et antal variable tages i betragtning. Disse variabler indbefatter: den samlede mængde og overfladeareal implantationsstedet, de mekaniske egenskaber af vævet, og vævet mikromiljø, som omfatter mængden af ​​vaskularisering og komponenterne i den ekstracellulære matrix. Derfor skal de materialer der anvendes til at levere disse celler være biokompatible med en defineret kemisk sammensætning under opretholdelse af en mekanisk styrke, som efterligner værtsvævet. Disse materialer skal også være permeabel for oxygen og næringsstoffer for at give en gunstig mikromiljø for celler til at vedhæfte og proliferere. Chitosan, en kationisk polysaccharid med fremragende bioforligelighed, kan let modificeres kemisk og har en høj affinitet til at binde med in vivo macromolecules 4-5. Chitosan efterligner glycosaminoglycan del af den ekstracellulære matrix, således at den fungerer som et substrat for celleadhæsion, migrering og proliferation. I denne undersøgelse har vi anvender chitosan i form af mikrosfærer til levering adipøst afledte stamceller (ASC) i en collagen baseret tredimensional stillads 6. En ideel celle-til-mikrosfære-forholdet blev bestemt med hensyn til inkuberingstid og celledensitet på at opnå maksimale antal celler, der kan indlæses. Når ASC udsås på chitosanmikrosfærer (CSM), er de indlejret i en collagen stillads og kan opretholdes i kultur i længere perioder. Sammenfattet tilvejebringer denne undersøgelse en fremgangsmåde til nøjagtig levering stamcellerne i en tredimensional biomateriale stillads.

Watch this Scientific Journal Video about Constructing a Collagen Hydrogel for the Delivery of Stem Cell-loaded Chitosan Microspheres at JoVE.com

Constructing a Collagen Hydrogel for the Delivery of Stem Cell-loaded Chitosan Microspheres

En stor hurdle i de nuværende stamceller er at bestemme den mest effektive metode til at levere disse celler til værtsvævene. Her beskriver vi en chitosan-baserede leveringsmetode som er effektiv og enkel fremgangsmåde, samtidig med at adipøst afledte stamceller for at bevare deres multipotency.

Konstruktion af et Collagen hydrogel til levering af stamcelle-loaded chitosanmikrosfærer

Multipotent stem cells have been shown to be extremely useful in the field of regenerative medicine1-3. However, in order to use these cells effectively ...

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Biomimetic enamel reconstruction is a significant topic in material science and dentistry as a novel approach for the treatment of dental caries or ...

Microscale Vortex-assisted Electroporator for Sequential Molecular Delivery

Individuel Vortex-assisteret elektroporator til sekventiel Molekylær Levering

Electroporation has received increasing attention in the past years, because it is a very powerful technique for physically introducing non-permeant ...

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Regeneration of lost myocardium is an important goal for future therapies because of the increasing occurrence of chronic ischemic heart failure and the ...

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