Name: tissue engineering of the intestine in a murine model
Uploaded: 2012-12-01T14:00:00
Description: Watch this Scientific Journal Video about Tissue Engineering of the Intestine in a Murine Model at JoVE.com

Tracy C. Grikscheit, Erik R. Barthel, og Frédéric G. Sala støttes av California Institute for regenerativ medisin (CIRM), Grant tall RN2-00946-1 (TCG) og TG2-01168 (ERB, FGS). Allison L. Speer er en forening av University kirurger Ethicon lærd. Yashuhiro Torashima er finansiert av et barnesykehus i Los Angeles Saban Forskning Career Development Fellowship.

1. Organoid Units Forberedelse <ol><li> Instrumenter som passer for mus disseksjon (saks og tang) bør steriliseres ved autoklavering. </li><li> Humant avlive donor mus i henhold til lokale IACUC protokoller. Sørge for at dyret er dødt før du fortsetter. </li><li> Foreta en midtlinjesnitt å få tilgang til bukhulen. Hud flaps kan bli reflektert som nødvendig for å forbedre eksponeringen. </li><li> Eviscerate tynntarmen og dele det bare distalt for ligament av Treitz. Separer tynntarmen fra sin mesentery hjelp sk...

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Vi presenterer en protokoll for å produsere vev-konstruert tarmen hos mus ved hjelp av en organoid enheter-on-stillas tilnærming. De mest kritiske trinn er de av organoid enheter forberedelse. Care må tas for å tilstrekkelig rent og mekanisk behandle vevet, men like omsorg må tas for ikke å overdigest eller overtriturate de organoid enheter etter fordøyelsen er utført (trinn 1.11). Hvis dette er gjort, kan de organoid enhetene reduseres til enkle celler, som kan gå tapt i supernatanten fra trinn 1,12, og er nep...

<table border="1"><tbody><tr><td> Navnet på reagensen </td><td> Selskapet </td><td> Katalognummer </td><td> Kommentarer (valgfritt) </td></tr><tr><td> HBSS </td><td> Gibco </td><td> 114170-112 </td><td></td></tr><tr><td> Antibiotika-antimycotic 100X </td><td> Invitrogen </td><td> 15240-062 </td><td></td></tr><tr><td> Dispase </td><td> Gibco </td><td> 17105-041 </td><td></td></tr><tr><td> Collagenase Type 1 </td><td> Worthington </td><td> LS004194 </td><td></td></tr><tr><td> DMEM Høy Glukose 1X </td><td> Gibco </td><td> 11995-065 </td><td></td></tr><tr><td> Varmeinaktivert FBS </td><td> Invitrogen </td><td> 16140-071 </td><td></td></tr><tr><td> Biofelt 100% PGA </td><td> Concordia Medical </td><td> FELT01-1005 </td><td> For polymer forberedelse som i Ref.4 </td></tr><tr><td> Poly-L-melkesyre </td><td> Durect </td><td> B6002-1 </td><td> For polymer forberedelse som i Ref. 4 </td></tr><tr><td> Type I kollagen, rotte hale </td><td> Sigma-Aldrich </td><td> C3867-1VL </td><td> For polymer forberedelse som i Ref. 4 </td></tr><tr><td> Ketoprofen 100 mg / ml </td><td> Fort Dodge Animal Health </td><td> 71-KETOI-100-50 </td><td></td></tr><tr><td> LabDiet 5001 gnager chow </td><td> LabDiet </td><td> 5001 </td><td></td></tr><tr><td> Septra 200 mg / 40 mg pr 5 ml, USP </td><td> Hi-Tech Pharmacal </td><td> 50383-824-16 </td><td></td></tr><tr><td> Isofluran, USP </td><td> Phoenix Pharmaceuticals </td><td> 57319-507-06 </td><td></td></tr></tbody></table>

tissue engineering of the intestine in a murine model

Vev-konstruert tynntarmen (Tesi) har med hell blitt brukt til å redde Lewis rotter etter massive tynntarmen reseksjon, noe som resulterer i retur til preoperativ vekter innen 40 dager. 1 Hos mennesker kan massive tynntarm reseksjon resultere i kort tarm-syndrom, en funksjonell malabsorptive staten som confers betydelig sykelighet, dødelighet og helsekostnader inkludert parenteral ernæring avhengighet, leversvikt og cirrhose, og behovet for multivisceral organtransplantasjon. 2 I denne artikkelen beskriver vi og dokumentere vår protokoll for å skape vev-konstruert tarmen i en mus modell med et flercellet organoid enheter-on-stillaset tilnærming. Organoid enheter er flercellede tilslag som stammer fra tarmen som inneholder både slimhinnene og mesenchymale elementer, 3 forholdet mellom som bevarer intestinal stamcelleforskningen nisje. 4 I pågående og fremtidig forskning, overgangen med teknikken vår imus vil gi rom for undersøkelse av prosessene under Tesi formasjon ved å utnytte de transgene verktøy tilgjengelig i denne arten. 5 Tilgjengeligheten av immunsupprimerte muselinjer vil også tillate oss å bruke teknikken til menneskelig intestinal vev og optimalisere dannelsen av menneskelig Tesi som mus xenograft før sin overgang til mennesker. Vår metode benytter Good Manufacturing Practice (GMP) reagenser og materialer som allerede er godkjent for bruk hos mennesker, og derfor gir en betydelig fordel i forhold tilnærminger som er avhengige decellularized animalsk vev. Det endelige målet med denne metoden er dens oversettelse til mennesker som regenerativ medisin terapeutisk strategi for kort tarm-syndrom.

Figur 1 viser en generell skjema for protokollen dokumentert her. Sluttresultatet av denne protokollen er en globus eller sfærisk struktur av vev-konstruert murine tarmen med en lumen, slimhinner, submucosa, og omkringliggende muskularis. Figur 2A viser en typisk verden i forhold til en start polymer stillas. Figur 2B viser samme konstruksjon kraftig bivalved å avsløre dens lumen. Figur 3 viser en hematoxylin / eosin-beiset parafin-montert tverrsnitt...

Watch this Scientific Journal Video about Tissue Engineering of the Intestine in a Murine Model at JoVE.com

Tissue Engineering of the Intestine in a Murine Model

Denne artikkelen og den medfølgende videoen presenterer vår protokoll for å generere vev-konstruert tarmen hos mus ved hjelp av en organoid enheter-on-stillas tilnærming.

Tissue Engineering av tarmen i en murine modell

Tissue-engineered small intestine (TESI) has successfully been used to rescue Lewis rats after massive small bowel resection, resulting in return to ...

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Bioengineering

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