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 er understøttet af California Institute til regenerativ medicin (CIRM), tilskud numre RN2-00.946-1 (TCG) og TG2-01.168 (ERB, FGS). Allison L. Speer er en Society of University Surgeons Ethicon lærd. Yashuhiro Torashima er finansieret af et børnehospital Los Angeles Saban Institute Research Career Development Fellowship.

1. Organoid Units Forberedelse <ol><li> Instrumenter egnede til muse-dissektion (saks og pincet) bør steriliseres ved autoklavering. </li><li> Humant aflive donormus efter lokale IACUC protokoller. Sørg for, at dyret er dødt, før du fortsætter. </li><li> Foretag en midtlinjeincision for at få adgang til bughulen. Hudlapper kan reflekteres efter behov for at forbedre eksponering. </li><li> Eviscerate tyndtarmen og dividere det bare distalt for ligament af Treitz. Adskil tyndtarmen fra sit mesenterium med skarp og...

<ol>
	<li>Grikscheit, T. C., Siddique, A., Ochoa, E. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue-engineered+small+intestine+improves+recovery+after+massive+small+bowel+resection.">Tissue-engineered small intestine improves recovery after massive small bowel resection.</a> Ann. Surg. 240, 748-754 (2004).</li><li>Wales, P. W., Christison-Lagay, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short+bowel+syndrome:+epidemiology+and+etiology.">Short bowel syndrome: epidemiology and etiology.</a> Sem. Ped. Surg. 19, 3-9 (2010).</li><li>Evans, G. S., Flint, N., Somers, A. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+of+a+method+for+the+preparation+of+rat+intestinal+epithelial+cell+primary+cultures.">The development of a method for the preparation of rat intestinal epithelial cell primary cultures.</a> J. Cell Sci. 101, 219-231 (1992).</li><li>Sala, F. G., Matthews, J. A., Speer, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+multicellular+approach+forms+a+significant+amount+of+tissue-engineered+small+intestine+in+the+mouse.">A multicellular approach forms a significant amount of tissue-engineered small intestine in the mouse.</a> Tiss. Eng. Part A. 17, 1841-1850 (2011).</li><li>Speer, A. L., Sala, F. G., Matthews, J. A., Grikscheit, T. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+tissue-engineered+stomach+demonstrates+epithelial+differentiation.">Murine tissue-engineered stomach demonstrates epithelial differentiation.</a> J. Surg. Res. 171, 6-14 (2011).</li><li>Haxhija, E. Q., Yang, H., Spencer, A. U., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+epithelial+cell+proliferation+is+dependent+on+the+site+of+massive+small+bowel+resection.">Intestinal epithelial cell proliferation is dependent on the site of massive small bowel resection.</a> Pediatr. Surg. Int. 23, 379-390 (2007).</li><li>Zhao, L., Cheng, Z., Dhall, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+corrective+pullthrough+surgery+in+a+mouse+model+of+Hirschsprung's+disease.">A novel corrective pullthrough surgery in a mouse model of Hirschsprung's disease.</a> J. Pediatr. Surg. 44, 759-766 (2009).</li><li>Petrosyan, M., Guner, Y. S., Williams, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+concepts+regarding+the+pathogenesis+of+necrotizing+enterocolitis.">Current concepts regarding the pathogenesis of necrotizing enterocolitis.</a> Ped. Surg. Int. 25, 309-318 (2009).</li><li>Shew, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+concerns+in+malrotation+and+midgut+volvulus.">Surgical concerns in malrotation and midgut volvulus.</a> Ped. Radiol. 39, S167-S171 (2009).</li><li>Sampietro, G. M., Corsi, F., Maconi, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prospective+study+of+long-term+results+and+prognostic+factors+after+conservative+surgery+for+small+bowel+Crohn's+disease.">Prospective study of long-term results and prognostic factors after conservative surgery for small bowel Crohn's disease.</a> Clin. Gastroenterol. Hepatol. 7, 183-191 (2009).</li><li>Klempnauer, J., Grothues, F., Bektas, H., Pichlmayr, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+results+after+surgery+for+acute+mesenteric+ischemia.">Long-term results after surgery for acute mesenteric ischemia.</a> Surgery. , 121-239 (1997).</li><li>Fitzgibbons, S. C., Jones, B. A., Hull, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+biopsy-proven+parenteral+nutrition-associated+liver+fibrosis+and+biochemical+cholestasis+in+children+with+short+bowel+syndrome.">Relationship between biopsy-proven parenteral nutrition-associated liver fibrosis and biochemical cholestasis in children with short bowel syndrome.</a> J. Ped. Surg. 45, 95-99 (2010).</li><li>Spencer, A. U., Kovacevich, D., McKinney-Barnett, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pediatric+short+bowel+syndrome:+the+cost+of+comprehensive+care.">Pediatric short bowel syndrome: the cost of comprehensive care.</a> Am. J. Clin. Nutr. 88, 1552-1559 (2008).</li><li>Kato, T., Tzakis, A. G., Selvaggi, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+and+multivisceral+transplantation+in+children.">Intestinal and multivisceral transplantation in children.</a> Ann. Surg. 243, 756-766 (2006).</li><li>Reyes, J., Bueno, J., Kocoshis, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+status+of+intestinal+transplantation+in+children.">Current status of intestinal transplantation in children.</a> J. Ped. Surg. 33, 243-254 (1998).</li></ol>

Vi præsenterer en protokol til fremstilling af vævs-manipuleret tarmen hos mus ved hjælp af en organoid enheder-on-stillads tilgang. De mest kritiske trin er sådanne med organoid enheder præparatet. Der skal udvises forsigtighed for tilstrækkeligt rene og mekanisk behandle væv, men lige man skal passe på ikke at overdigest eller overtriturate de organoid enheder efter fordøjelsen udføres (trin 1,11). Hvis dette sker, kan de organoid enheder reduceres til enkelte celler, som kan gå tabt i supernatanten af ​?...

<table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer (valgfrit) </td></tr><tr><td> HBSS </td><td> Gibco </td><td> 114170-112 </td><td></td></tr><tr><td> Antibiotika-antimykotisk 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> Kollagenase type 1 </td><td> Worthington </td><td> LS004194 </td><td></td></tr><tr><td> DMEM High Glucose 1X </td><td> Gibco </td><td> 11995-065 </td><td></td></tr><tr><td> Varmeinaktiveret 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> Til fremstilling af polymer som i ref.4 </td></tr><tr><td> Poly-L-mælkesyre </td><td> Durect </td><td> B6002-1 </td><td> Til fremstilling af polymer som i ref. 4 </td></tr><tr><td> Type I kollagen, rotte hale </td><td> Sigma-Aldrich </td><td> C3867-1VL </td><td> Til fremstilling af polymer 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 gnaverfoder </td><td> LabDiet </td><td> 5001 </td><td></td></tr><tr><td> Septra 200 mg / 40 mg per 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

Tissue-engineered tyndtarmen (Tesi) har med held blevet anvendt til at redde Lewis-rotter efter massiv resektion af tyndtarmen, hvilket medfører tilbagevenden til præoperative vægte inden for 40 dage. En Hos mennesker kan massiv resektion af tyndtarmen resultere i korttarmssyndrom, en funktionel malabsorptive stat, der giver høj sygelighed, dødelighed og udgifter til sundhedsvæsenet, herunder parenteral ernæring afhængighed, leversvigt og skrumpelever, og behovet for multivicerale organtransplantation. 2 I dette papir, vi beskrive og dokumentere vores protokol for at skabe væv-manipuleret tarmen i en musemodel med en multicellulær organoid enheder-on-skelet fremgangsmåde. Organoid enheder er flercellede aggregater stammer fra tarmen, der indeholder både mucosale og mesenchymal elementer, 3 forholdet mellem der bevarer den intestinale stamceller niche. 4 I igangværende og fremtidige forskning, overgangen fra vores teknik tilmus vil give mulighed for undersøgelse af de processer, der er involveret i løbet af Tesi dannelse ved at udnytte de transgene tilgængelige værktøjer i denne art. 5 Tilgængeligheden af immunkompromitterede musestammer vil også tillade os at anvende teknikken til menneskelig tarmvæv og optimere dannelsen af menneskelig Tesi som en mus xenograft før dens overgang til mennesker. Vores metode anvender god fremstillingspraksis (GMP) reagenser og materialer, der allerede er godkendt til brug i humane patienter, og derfor giver en betydelig fordel i forhold til tilgange, der er baseret på decellulariseret dyrevæv. Det endelige mål med denne metode er dens translation til mennesker som en regenerativ medicin terapeutisk strategi for korttarmssyndrom.

Figur 1 viser et samlet skema for protokollen dokumenteret her. Slutresultatet af denne protokol er en globus eller sfærisk struktur af vævs-engineering murint tarmen med et lumen, slimhinder, submucosa, og omgivende muscularis. Figur 2A viser en typisk globus i forhold til et udgangspunkt polymerskelet. Figur 2B viser den samme konstruktion kraftigt bivalved at afsløre dets lumen. Figur 3 viser et hematoxylin / eosin-stained paraffin-monteret tværs...

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 artikel og den ledsagende video præsentere vores protokol for frembringelse tissue-engineered tarmen hos mus under anvendelse af en organoid enheder-on-skelet fremgangsmåde.

Tissue Engineering af tarmen i en murin model

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

Research

JoVE Journal

Bioengineering

Postproduction Processing of Electrospun Fibres for Tissue Engineering

Postproduktion Behandling af Electrospun Fibre til Tissue Engineering

Electrospinning is a commonly used and versatile method to produce  scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo ...

Engineering Skeletal Muscle Tissues from Murine Myoblast Progenitor Cells and Application of Electrical Stimulation

Engineering skeletmuskel Væv fra murine myoblast progenitorceller og anvendelse af elektrisk stimulering

Engineered muscle tissues can be used for several different purposes, which include the production of tissues for use as a disease model in vitro, e.g. to ...

Design of a Biaxial Mechanical Loading Bioreactor for Tissue Engineering

Design af en Biaksial Mekanisk Loading bioreaktor for Tissue Engineering

We  designed a loading device that is capable of applying uniaxial or biaxial  mechanical strain to a tissue engineered biocomposites fabricated for  ...

Tissue Engineering of a Human 3D in vitro Tumor Test System

Tissue Engineering of a Human 3D in vitro Tumor Test System

Tissue Engineering af et menneske 3D<em&gt; In vitro</em&gt; Tumor Test System

Cancer is one of the leading causes of death worldwide.  Current therapeutic strategies are predominantly developed in 2D culture  systems, which ...

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

Generation af en tre-dimensionel fuld tykkelse Skin Equivalent og Automated Anskydning

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

Production, Characterization and Potential Uses of a 3D Tissue-engineered Human Esophageal Mucosal Model

Produktion, karakterisering og potentielle anvendelse af en 3D-Tissue-manipuleret humant Esophageal Mucosal Model

The incidence of both esophageal adenocarcinoma and its precursor, Barrett&#8217;s Metaplasia, are rising rapidly in the western world. Furthermore ...

Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration

Engineering 3D Cellularized kollagengeler for Vascular Tissue Regeneration

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. ...

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

Tissue Engineering af Intrinsic Vaskularisering i en<em&gt; In vivo</em&gt; Tissue Engineering Chamber

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

The Arteriovenous (AV) Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

The Arteriovenous AV Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

Den arteriovenøse (AV) Loop i en lille Animal Model at studere angiogenese og vaskulariseret Tissue Engineering

A functional blood vessel network is a prerequisite for the survival and growth of almost all tissues and organs in the human body. Moreover, in ...

Treatment of Ligament Constructs with Exercise-conditioned Serum: A Translational Tissue Engineering Model

Behandling af ligamentkonstruktioner med træningskonditioneret serum: en translational tissue engineering model

In vitro experiments are essential to understand biological mechanisms; however, the gap between monolayer tissue culture and human physiology is large, ...