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

Die Autoren sind dankbar, dass Prof. Dr. S.E.R Hovius, Dr. M.A.M. Mureau und alle Chirurgen der Abteilung für plastische Chirurgie für die Sammlung des subkutanen Fettgewebes. P. Sukho wird durch einen Zuschuss aus den Niederlanden Fellowship-Programm (NFP-12/435), während der Durchführung der Studie unterstützt. Y.M. Bastiaansen-Jenniskens wird unterstützt durch Zuschüsse aus niederländischen Arthritis Foundation (LP11).

Subkutane abdominalen Fettgewebe erhielt von menschlichen Spendern mit Genehmigung der medizinischen Ethik-Ausschuß (#MEC-2014-092), Erasmus MC University Medical Center, Rotterdam, Niederlande. Alle Tierversuche stimmten mit den ethischen Komitees der Tier experimentieren, Erasmus MC University Medical Center, Rotterdam, Niederlande (133-14-01).
1. menschliche ASCs Isolation und Kultur
<ol><li>Bereiten Sie 3 mL Isolierung Medium für 1 g des Fettgewebes. Niedrige Glukose Mischung Dulbecco'...

<ol>
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Anastomosen Leckage ist die am meisten Schwerwiegendes unerwünschtes Ereignis nach Kolon-Resektion mit eine primäre Anastomose. Optimale Techniken zur Vermeidung von Anastomosen Störung und Leckage fehlen noch. Lokale Anwendung eines Arrays von Biomaterialien wurde mit unterschiedlichen Ergebnisse25,26,27durchgeführt. Zelltherapien soll Gewebereparatur von Gewebeersatz oder die Stimulation der lokalen Heilung durch Parakrine...

Partielle Colectomy mit eine primäre Anastomose ist eine häufig durchgeführte Operation, die für viele Erkrankungen des Dickdarms wie Darmkrebs, Morbus Crohn und Divertikulitis1,2getan werden kann. Die am meisten gefürchtete Komplikation nach kolorektalen Anastomose Anastomosen Leckage3ist. Obwohl mehrere Risikofaktoren im Zusammenhang mit Anastomosen Lecks identifiziert wurden, sind Lösungen zur Vermeidung von Leckagen Unkown4,5.
Fettgewebe gewonnenen Stromazellen Zellen (ASC) werden entzündungshemmende und trophischen Eigenschaften6,7, wodurch diese vielversprechende Kandidaten für regenerative Therapien8Zellen zugeordnet. Die Wirksamkeit des ASC, die Heilung des Gewebes zu fördern wurde in verschiedenen Geweben wie z. B. des Herzmuskels, Haut und Speiseröhre9,10,11,12,13gezeigt. Allerdings gibt es einige Berichte über den Einsatz von ASC, intestinale Heilung zu fördern. Lokale Transplantation von ASC, experimentelle kolorektalen Anastomosen über ASC-beschichtete Biosutures oder serosal Injektionen von ASC zeigte sich entweder keine Besserung bei der Heilung14 nicht verhindern, dass Anastomosen Leckage trotz ein günstigeres Anastomosen 15zu heilen.
Lokale Transplantation des ASC in der Schwebe oder in Kombination mit Biomaterialien möglicherweise unzureichend Zelle Zurückbehaltungs- oder eine unzureichende Verteilung der transplantierten Zellen11zugeordnet. Zelle Blatt Technologie16,17 bietet eine innovative Methode der ASC Lieferung18,19. Daher wurde in einer früheren Studie, ein neuartiger Ansatz vorgeschlagen, in welche ASCs organisiert in einer Zelle auf experimentelle kolorektalen Anastomose20Blatt angewendet werden kann. Diese Studie zeigte, dass ASC Blatt Transplantation erfolgreich bei der Verringerung der kolorektalen Anastomose Leckage nach teilweiser Colectomy in einem Rattenmodell ist. Dieser Artikel berichtet ASC Blatt Vorbereitung und chirurgische Transplantation Technik.

<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

Anastomosen Leckage ist eine katastrophale Komplikation nach Kolorektale Chirurgie. Obwohl aktuelle Methoden zur Leckage Prävention unterschiedlicher klinischer Wirksamkeit haben, sind sie bis jetzt unvollkommene Lösungen. Stammzell-Therapie mit ASC Sheets könnte eine Lösung für dieses Problem bieten. ASC gelten als aussichtsreiche Kandidaten für die Förderung der Gewebe, die Heilung aufgrund ihrer trophischen und immunmodulatorische Eigenschaften. Hier bieten wir Methoden, um High-Density-ASC Blätter zu produzieren, die auf einem kolorektalen Anastomose in einem Rattenmodell der Leckage reduzieren verpflanzt werden. ASC gebildet Zelle Blätter in Thermo-responsive Kultur-Gerichte, die leicht abgestellt werden können. Am Tag der Transplantation erfolgte eine teilweise Colectomy mit einem 5-Naht kolorektalen Anastomose. Tiere wurden sofort mit 1 ASC Blatt pro Ratte transplantiert. ASC Blätter eingehalten spontan die Anastomose ohne Klebstoff, Naht oder irgendwelche Biomaterial. Tiergruppen wurden geopfert, 3 bis 7 Tagen nach der Operation. Im Vergleich zu transplantierten Tieren, lag die Inzidenz von Anastomosen Abszesse und Leckage in Kontrolltieren. In unserem Modell war die Transplantation von ASC Blätter nach kolorektalen Anastomose erfolgreich und mit einer geringeren Leckrate verbunden.

Ein Flussdiagramm der Studie Darstellung ASC Blatt Kultur und das Verfahren der Kolon-Resektion und Anastomose ist in Abbildung 1dargestellt. Abbildung 2 zeigt ASC Blatt mikroskopische Morphologie und das makroskopische Erscheinungsbild der ASC-Blatt während und nach der Ablösung. Abbildung 3 zeigt die verschiedenen Stufen des ASC Blatt ablösen und Transplantation. Abbildung 4...

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

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

Transplantation von Fettgewebe-Derived Stem Cell Blatt, Leckage nach teilweiser Colectomy in einem Rattenmodell zu reduzieren

Die Vorbereitung und die Transplantation von abgeleitet Fettgewebe Stammzellen (ASC) Blätter auf unzureichend genähten kolorektalen Anastomosen in einem Rattenmodell wird vorgestellt. Diese neuartige Anwendung zeigt, dass ASC Blätter kolorektalen Anastomose Leckage reduziert werden können.

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

Orthotopen Aorten-Transplantation: Ein Rattenmodell die Entwicklung von chronischen Vaskulopathie Study

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

Forschung

Medizin

A Mouse Model of in Utero Transplantation

A Mouse Model of in Utero Transplantation

Einem Mausmodell der 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

Stem Cell Transplantation in einem In-vitro- Simulierte Ischämie / Reperfusion-Modell

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

Chirurgische Verfahren für eine Ratte Modell Teilweise Lebertransplantation mit arteriellen Rekonstruktion

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 Stem Cell Transplantation in Experimental Gequetscht Modell von Rückenmarksverletzungen

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

Schaffung und Transplantation von einem Fettgewebe-derived Stem Cell (ASC) Blatt in einem diabetischen Wundheilung Modell

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-sheet Transplantation Stricture nach Erweiterte Speiseröhren Endoskopische Submukosa Dissektion zu verhindern

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

Intrasplenic Transplantation von Leberzellen nach teilweiser Hepatektomie in NOD. SCID-Mäuse

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 Arterialized Ratte teilweise Leber Transplantation mit einem in Vivo Schiff-orientierte 70 % Hepatektomie

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

Reduzierte Komplikationen nach arterieller Wiederverbindung in einem Rattenmodell der orthotopischen Lebertransplantation

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