Name: transplantation of adipose tissue-derived stem cell sheet to reduce leakage after partial colectomy in a rat model
Uploaded: 2018-08-11T12:30:00
Description: Watch this Scientific Journal Video about Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model at JoVE.com

Les auteurs sont reconnaissants aux Prof. Dr. S.E.R. Hovius, Dr m.a.M. Mureau et tous les chirurgiens du département de chirurgie plastique de la collection du tissu adipeux sous-cutané. Sukho P. est soutenu par une subvention du programme The Netherlands Fellowship (NFP-12/435), pendant le déroulement de l’étude. Y.M. Bastiaansen-Jenniskens est pris en charge par des subventions de la Fondation néerlandaise de l’arthrite (LP11).

Du tissu adipeux sous-cutané abdominal proviennent de donneurs humains avec l’approbation du Comité d’éthique médicale (#MEC-2014-092), Erasmus MC University Medical Center, Rotterdam, Pays-Bas. Toutes les expériences animales ont été approuvées par le Comité éthique d’expérimentation animale, centre médical universitaire Erasmus MC, Rotterdam, Pays-Bas (133-14-01).
1. culture et l’isolement de l’ASCs humaines
<ol><li>Préparer 3 mL de milieu d’isolement pour 1 g de t...

<ol>
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Cell Harvesting by Temperature Reduction Available from: <a target="_blank" href="https://tools.thermofisher.com/content/sfs/brochures/AppNote-Nunc-UpCell-N20230.pdf">https://tools.thermofisher.com/content/sfs/brochures/AppNote-Nunc-UpCell-N20230.pdf</a> (2008)</li><li>Wu, Z., 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=Colorectal+anastomotic+leakage+caused+by+insufficient+suturing+after+partial+colectomy:+a+new+experimental+model.">Colorectal anastomotic leakage caused by insufficient suturing after partial colectomy: a new experimental model.</a> Surg Infect (Larchmt). 15 (6), 733-738 (2014).</li><li>Wu, Z., 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=Reducing+colorectal+anastomotic+leakage+with+tissue+adhesive+in+experimental+inflammatory+bowel+disease.">Reducing colorectal anastomotic leakage with tissue adhesive in experimental inflammatory bowel disease.</a> Inflamm Bowel Dis. 21 (5), 1038-1046 (2015).</li><li>Wu, Z., 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=Reducing+anastomotic+leakage+by+reinforcement+of+colorectal+anastomosis+with+cyanoacrylate+glue.">Reducing anastomotic leakage by reinforcement of colorectal anastomosis with cyanoacrylate glue.</a> Eur Surg Res. 50 (3-4), 255-261 (2013).</li><li>Aysan, E., Dincel, O., Bektas, H., Alkan, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polypropylene+mesh+covered+colonic+anastomosis.+Results+of+a+new+anastomosis+technique.">Polypropylene mesh covered colonic anastomosis. 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Une fuite anastomotique est l’évènement perturbateur plus grave après la résection du côlon avec une anastomose primaire. Techniques optimales pour éviter les fuites et rupture anastomotique manquent encore. L’application locale d’un tableau des biomatériaux a été réalisée, avec des résultats25,26,27. Le but des thérapies cellulaires est de faciliter la réparation des tissus de remplacement de tissus ou de la...

Colectomie partielle avec une anastomose primaire est une intervention couramment pratiquée qui peut être faite pour de nombreuses maladies touchant le côlon telles que le cancer colorectal, maladie de Crohn et diverticulite1,2. Le plus redoutable complication après que anastomose colorectal est fuite anastomotique3. Bien que plusieurs facteurs de risque associés aux fuites anastomotiques ont été identifiées, des solutions pour prévenir les fuites restent inconnu4,5.
Cellules stromales dérivées de tissu adipeux (ASCs) sont associés aux propriétés anti-inflammatoires et trophiques6,7, qui rend ces cellules promettant des candidats pour les thérapies régénératives8. L’efficacité des CRA pour favoriser la guérison des tissus a été montrée dans différents tissus comme le muscle cardiaque, peau et oesophage9,10,11,12,13. Cependant, il y a peu de rapports sur l’utilisation des CRA pour favoriser la guérison intestinale. Locale transplantation des CRA à expérimentales anastomoses colorectal via biosutures ASC-enduit ou injections séreux des CRA ont montré ni aucune amélioration dans la guérison14 ou n’a pas empêché les fuites anastomotiques malgré un résultat plus favorable anastomotique 15la guérison.
Locale transplantation des CRA en suspension ou combinée avec des biomatériaux peut-être être associés à une rétention cellulaire insuffisante ou une répartition inadéquate des cellules transplantées11. Cellules feuille technologie16,17 propose une méthode innovante de NCP livraison18,19. Par conséquent, dans une étude antérieure, une nouvelle approche a été proposée dans lequel ASCs organisés dans une cellule feuille pourrait être appliquée sur expérimental colorectaux anastomose20. Cette étude a démontré que transplantation feuille ASC a réussi à réduire les fuites colorectaux anastomose après colectomie partielle dans un modèle de rat. Cet article rapporte ASC feuille préparation et technique de la transplantation chirurgicale.

<table border="0" cellpadding="0" cellspacing="0" width="689">
	<tbody>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">LG DMEM</td>
			<td style="width:115px;">Gibco, Life technologies</td>
			<td style="width:191px;">22320022</td>
			<td style="width:167px;">ASC isolation and culture</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Collagenase type I</td>
			<td style="width:115px;">Gibco, Life technologies</td>
			<td style="width:191px;">17100-01</td>
			<td style="width:167px;">ASC Isolation</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Bovine Serum Albumin</td>
			<td style="width:115px;">Sigma-Aldrich</td>
			<td style="width:191px;">A9418</td>
			<td style="width:167px;">ASC Isolation</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Fetal bovine serum</td>
			<td style="width:115px;">Gibco, Life technologies</td>
			<td style="width:191px;">10270-106</td>
			<td style="width:167px;">FBS, ASC isolation and culture</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">3% acetic acid with methylene blue</td>
			<td style="width:115px;">Stemcell Technologies</td>
			<td style="width:191px;">7060</td>
			<td style="width:167px;">ASC Isolation</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Gentamicin</td>
			<td style="width:115px;">Gibco, Life technologies</td>
			<td style="width:191px;">15750-037</td>
			<td style="width:167px;">ASC isolation and culture</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Ampothericin B&#160;</td>
			<td style="width:115px;">Gibco, Life technologies</td>
			<td style="width:191px;">15290-018</td>
			<td style="width:167px;">ASC isolation and culture</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:216px;">Shaker (Gallenkamp Environmental Shaker Model 10X 400)</td>
			<td style="width:115px;">Akribis Scientific</td>
			<td style="width:191px;">F240</td>
			<td style="width:167px;">ASC Isolation</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Centrifuge&#160;</td>
			<td style="width:115px;">Hettichlab&#160;</td>
			<td style="width:191px;">Rotina380</td>
			<td style="width:167px;">ASC isolation and culture</td>
		</tr>
		<tr height="62">
			<td height="62" style="height:62px;width:216px;">Phosphate buffer saline&#160;</td>
			<td style="width:115px;">Gibco, Life technologies</td>
			<td style="width:191px;">14190-094</td>
			<td style="width:167px;">PBS, ASC isolation and culture</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ascorbic acid-2-phosphate</td>
			<td style="width:115px;">Sigma-Aldrich</td>
			<td style="width:191px;">A8960</td>
			<td style="width:167px;">ASC isolation and culture</td>
		</tr>
		<tr height="37">
			<td height="37" style="height:37px;width:216px;">Human recombinant fibroblast growth factor 2&#160;</td>
			<td style="width:115px;">AbD Serotec</td>
			<td style="width:191px;">AF-100-15</td>
			<td style="width:167px;">FGF2, ASC isolation and culture</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Trypsin EDTA</td>
			<td style="width:115px;">Gibco, Life technologies</td>
			<td style="width:191px;">25200-056</td>
			<td style="width:167px;">ASC culture</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Dimethyl sulfoxide</td>
			<td style="width:115px;">Sigma-Aldrich</td>
			<td style="width:191px;">D2650-5x</td>
			<td style="width:167px;">DMSO, ASC freezing</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Thermo-responsive culture dishes</td>
			<td style="width:115px;">Nunc Thermo scientific</td>
			<td style="width:191px;">174904</td>
			<td style="width:167px;">ASC sheet preperation</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Non-absorbable braided silk 4/0</td>
			<td style="width:115px;">B Braun</td>
			<td style="width:191px;">21151042</td>
			<td style="width:167px;">surgery</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Non-absorbable monofilament polyamide&#160; 8/0</td>
			<td style="width:115px;">B Braun</td>
			<td style="width:191px;">G1118170</td>
			<td style="width:167px;">surgery</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Absorbable braided polyglycolic acid 5/0</td>
			<td style="width:115px;">B Braun</td>
			<td style="width:191px;">C1048207</td>
			<td style="width:167px;">surgery</td>
		</tr>
	</tbody>
</table>

transplantation of adipose tissue-derived stem cell sheet to reduce leakage after partial colectomy in a rat model

Une fuite anastomotique est une complication désastreuse après chirurgie colorectale. Bien que les méthodes actuelles pour la prévention de fuites ont différents niveaux d’efficacité clinique, ils sont jusqu'à présent imparfaites solutions. Thérapie de cellules souches en utilisant des feuilles de l’ASC pourrait constituer une solution à ce problème. CRA est considérés comme des candidats aussi prometteurs pour la promotion des tissus de guérison en raison de leurs propriétés trophiques et immunomodulatrices. Ici, nous fournissent des méthodes pour produire des feuilles de ASC haute densités, qui sont transplantés sur une anastomose colorectal dans un modèle de rat pour réduire les fuites. CRA formé de feuilles de cellule dans les récipients de culture thermo sensible qui pourraient facilement être enlevés. Le jour de la transplantation, une colectomie partielle avec une anastomose colorectal 5-suture a été réalisée. Animaux ont été transplantés immédiatement avec 1 feuille de NCP par rat. Feuilles de l’ASC ont adhéré spontanément à l’anastomose sans colle, suture ou tout biomatériau. Groupes d’animaux ont été sacrifiés 3 et 7 jours après l’opération. Par rapport aux animaux transplantés, l’incidence d’abcès anastomotiques et la fuite était plus élevé chez les animaux témoins. Dans notre modèle, la transplantation de feuilles ASC après colorectaux anastomose était réussie et associé à un débit de fuite inférieur.

Un organigramme de cette étude décrivant les ASC fiche culture et la procédure de la résection du côlon et anastomose est illustré à la Figure 1. La figure 2 illustre la morphologie microscopique des ASC feuille et l’aspect macroscopique de la feuille de l’ASC pendant et après le détachement. La figure 3 montre les différentes étapes du détachement de feuille d’ASC et de la transpla...

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Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Transplantation de la feuille les cellules souches dérivées du tissu adipeux pour réduire les fuites après colectomie partielle dans un modèle de Rat

La préparation et la transplantation de feuilles (ASC) les cellules souches dérivé du tissu adipeux sur les anastomoses colorectal insuffisamment suturées dans un modèle de rat est présenté. Cette nouvelle application montre que feuilles de NCP peuvent réduire les fuites d’anastomose colorectal.

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Research

JoVE Journal

Medicine

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Orthotopique aortique transplantation: un modèle de rat pour étudier le développement de la vasculopathie chronique

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

Recherche

Médecine

A Mouse Model of in Utero Transplantation

A Mouse Model of in Utero Transplantation

Un modèle murin de In utero Transplantation

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus.  For example, in ...

Stem Cell Transplantation in an in vitro Simulated Ischemia/Reperfusion Model

Stem Cell Transplantation in an in vitro Simulated Ischemia/Reperfusion Model

Transplantation de cellules souches dans un In vitro Simulation d&#39;ischémie / reperfusion modèle

Stem cell transplantation protocols are finding their way into clinical practice1,2,3. Getting better results, making the protocols more robust, and ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Interventions chirurgicales pour un modèle de rat de transplantation hépatique orthotopique partielle avec reconstruction artérielle hépatique

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Neural Stem Cell Transplantation in Experimental Contusive Model of Spinal Cord Injury

Neural Transplantation de cellules souches dans la partie expérimentale Modèle contusion de la moelle épinière des blessures

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can ...

Creation and Transplantation of an Adipose-derived Stem Cell (ASC) Sheet in a Diabetic Wound-healing Model

Creation and Transplantation of an Adipose-derived Stem Cell ASC Sheet in a Diabetic Wound-healing Model

Création et la Transplantation d’une feuille adipeux dérivées de cellules souches (ASC) dans un modèle de cicatrisation diabétique

Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients ...

ADSC-sheet Transplantation to Prevent Stricture after Extended Esophageal Endoscopic Submucosal Dissection

ADSC-feuille Transplantation à prévenir Rétrécissement après oesophagienne prolongée Endoscopic dissection sous-muqueuse

In past years, the cell-sheet construct has spurred wide interest in regenerative medicine, especially for reconstructive surgery procedures. The ...

Intrasplenic Transplantation of Hepatocytes After Partial Hepatectomy in NOD.SCID Mice

Intra-splénique Transplantation d’hépatocytes après hépatectomie partielle en clin de œil. Souris SCID

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological ...

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Re-artérialisé Transplantation de foie partielle de Rat in vivo axée sur le navire 70 % hépatectomie

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

Complications réduites après reconnexion artérielle dans un modèle de rat de transplantation orthotopique de foie

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...