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

著者は、皮下脂肪組織のコレクションの外科の部門のすべての外科医、博士 M.A.M。 Mureau 教授博士 S.E.R. 活発に感謝しています。P. Sukho は、試験の実施中にオランダのフェローシップ プログラム (非営利団体-12/435) からの助成金によってサポートされます。Y. m. Bastiaansen-Jenniskens は、オランダの関節炎財団 (LP11) からの助成金によってサポートされます。

腹部皮下脂肪は医療倫理委員会 (#MEC-2014-092)、エラスムス MC 大学医療センター、ロッテルダム、オランダの承認を得て人間のドナーから得られました。すべての動物実験は、動物実験倫理委員会、エラスムス MC 大学医療センター、ロッテルダム、オランダ (133-14-01) によって承認されました。
1. 人間の Asc の分離と培養
<ol><li>脂肪の 1 グラムの分離培地 3 mL を準備します...

<ol>
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T., 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=Human+adipose+stem+cells+cell+sheet+constructs+impact+epidermal+morphogenesis+in+full-thickness+excisional+wounds.">Human adipose stem cells cell sheet constructs impact epidermal morphogenesis in full-thickness excisional wounds.</a> Biomacromolecules. 14 (11), 3997-4008 (2013).</li><li>Perrod, 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=Cell+Sheet+Transplantation+for+Esophageal+Stricture+Prevention+after+Endoscopic+Submucosal+Dissection+in+a+Porcine+Model.">Cell Sheet Transplantation for Esophageal Stricture Prevention after Endoscopic Submucosal Dissection in a Porcine Model.</a> PLoS One. 11 (3), e0148249 (2016).</li><li>Pascual, I., 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=Adipose-derived+mesenchymal+stem+cells+in+biosutures+do+not+improve+healing+of+experimental+colonic+anastomoses.">Adipose-derived mesenchymal stem cells in biosutures do not improve healing of experimental colonic anastomoses.</a> Brit J Surg. 95 (9), 1180-1184 (2008).</li><li>Yoo, J. H., 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=Adipose-tissue-derived+Stem+Cells+Enhance+the+Healing+of+Ischemic+Colonic+Anastomoses:+An+Experimental+Study+in+Rats.">Adipose-tissue-derived Stem Cells Enhance the Healing of Ischemic Colonic Anastomoses: An Experimental Study in Rats.</a> J Korean Soc Coloproctol. 28 (3), 132-139 (2012).</li><li>Okano, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+sheet+engineering.">Cell sheet engineering.</a> Rinsho Shinkeigaku. 46 (11), 795-798 (2006).</li><li>Chen, 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=Application+of+the+cell+sheet+technique+in+tissue+engineering.">Application of the cell sheet technique in tissue engineering.</a> Biomed Rep. 3 (6), 749-757 (2015).</li><li>Labbe, B., Marceau-Fortier, G., Fradette, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+sheet+technology+for+tissue+engineering:+the+self-assembly+approach+using+adipose-derived+stromal+cells.">Cell sheet technology for tissue engineering: the self-assembly approach using adipose-derived stromal cells.</a> Methods Mol Biol. 702, 429-441 (2011).</li><li>Sukho, P., 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=Adipose+Tissue-Derived+Stem+Cell+Sheet+Application+for+Tissue+Healing+In+Vivo:+A+Systematic+Review.">Adipose Tissue-Derived Stem Cell Sheet Application for Tissue Healing In Vivo: A Systematic Review.</a> Tissue Eng Part B-Re. , (2017).</li><li>Sukho, P., 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=Effects+of+adipose+stem+cell+sheets+on+colon+anastomotic+leakage+in+an+experimental+model:+Proof+of+principle.">Effects of adipose stem cell sheets on colon anastomotic leakage in an experimental model: Proof of principle.</a> Biomaterials. 140, 69-78 (2017).</li><li>Verseijden, F., 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=Angiogenic+capacity+of+human+adipose-derived+stromal+cells+during+adipogenic+differentiation:+an+in+vitro+study.">Angiogenic capacity of human adipose-derived stromal cells during adipogenic differentiation: an in vitro study.</a> Tissue Eng Pt A. 15 (2), 445-452 (2009).</li><li>Sukho, P., 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=Effect+of+Cell+Seeding+Density+and+Inflammatory+Cytokines+on+Adipose+Tissue-Derived+Stem+Cells:+an+in+Vitro+Study.">Effect of Cell Seeding Density and Inflammatory Cytokines on Adipose Tissue-Derived Stem Cells: an in Vitro Study.</a> Stem Cell Rev. 13 (2), 267-277 (2017).</li><li>. 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. Results of a new anastomosis technique.</a> Int J Surg. 6 (3), 224-229 (2008).</li><li>Suarez-Grau, J. 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縫合は、プライマリ吻合による結腸切除術後最も重篤な有害事象です。吻合部の中断や漏れを防ぐために最適な技術はまだ欠けています。生体材料の配列のローカル アプリケーションはさまざまな結果25,26,27と、実施されています。細胞療法の目的は、組織置換による組織修復やパラクリン分泌を介してローカル癒しの刺...

主な吻合を持つ部分的な結腸切除術は、大腸癌、クローン病、憩室炎1,2など大腸に影響を与える多くの病気のため行うことができます一般的に行わ手術です。大腸吻合は縫合3後、最も恐ろしい合併症です。吻合部リークに関連付けられているいくつかの危険因子が同定されている、不明4、5のまま漏洩防止ソリューションです。
脂肪組織由来の間質細胞 (Asc) は、抗炎症および栄養特性6、7、再生治療8候補者を約束これらの細胞を作るに関連付けられます。心臓の筋肉、皮膚、食道9,10、11,12,13など様々 な組織に組織の治癒を促進するため Asc の有効性を示した。ただし、腸治癒を促進する Asc の使用に関するほとんどのレポートがありません。ASC コーティング biosutures を介して実験的大腸吻合に Asc のローカル移植または Asc の漿膜注射癒しの14のいずれかなしの改善を示した吻合部をより好ましいにもかかわらず縫合を予防しなかった15をの癒し。
懸濁液の Asc のローカル移植併用または生体が不十分なセルの保持または移植細胞11の不十分な分布に関連付けられた可能性があります。細胞シート技術16,17 ASC 配信18,19の革新的な方法を提供しています。したがって、以前の研究では、新しいアプローチは実験的大腸吻合20で適用できるシートのセルで構成されている Asc を提案しました。この研究は、ASC シート移植がラットモデルにおける部分的な術後後大腸吻合の漏れを減らすに成功したことを示した。この記事は、ASC シート準備と外科的移植法を報告します。

<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

縫合は、大腸手術後悲惨な合併症です。漏洩防止のための現在の方法は、臨床的有効性の異なるレベルを持って、彼らは今不完全なソリューションまでです。ASC のシートを使用して幹細胞療法は、この問題への解決策を提供できます。Asc は、組織のための栄養と免疫調節特性治癒を促進するために有望な候補者と見なされます。ここでは、漏れを減らすためラットのモデル大腸吻合移植された高密度の ASC シートを生産するためのメソッドを提供します。Asc は、容易に切り離すことができる温度応答性培養皿の細胞シートを形成しました。移植の日、5 縫合大腸吻合による部分的な結腸切除術を行った.動物はラット ASC 1 枚入ですぐに定植。ASC シートは、任意の接着剤、縫合、または任意の生体材料なし吻合に自発的に付着しました。動物のグループが 3 と 7 日を犠牲になった術後。移植した動物に比べると、対照動物の吻合部膿瘍と漏れの発生率は高かった。私たちのモデル大腸吻合後 ASC シートの移植は成功し漏れ率が低いに関連付けられています。

ASC シート文化と結腸切除および吻合術の手順を描いたこの研究の流れは、図 1に示すです。図 2は、ASC シート顕微鏡的形態と中および剥離後 ASC シートの巨視的な外観を示しています。図 3は、ASC シート剥離・移植の別の手順を示します。図 4示します吻合と漏洩防止の ASC シー?...

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

ラットモデルにおける部分的な術後後の漏れを減らすために脂肪組織由来幹細胞シートの移植

準備・ ラットのモデル大腸吻合の縫合が不十分な上に脂肪組織由来幹細胞 (ASC) シートの移植が表示されます。この新しいアプリケーションは、ASC シートが大腸吻合の漏れを減らすことができますを示しています。

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

同所性大動脈移植：慢性血管障害の発生研究のラットモデル

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

A Mouse Model of in Utero Transplantation

A Mouse Model of in Utero Transplantation

のマウスモデル子宮内で移植

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

における幹細胞移植 in vitroで模擬虚血/再灌流モデル

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

肝動脈再建を用いた部分同所性肝移植のラットモデルのための外科的処置

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

脊髄損傷の実験的挫傷モデルにおける神経幹細胞移植

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

作成および糖尿病創傷治癒モデルにおける脂肪由来幹細胞 (ASC) シートの移植

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

拡張食道内視鏡的粘膜下層切開後に狭窄を防止するためのADSC-シート移植

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

うなずくにおける肝切除後肝脾内移植。SCID マウス

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

再門脈部分肝移植のラット体内血管指向 70% 肝切除

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

ラット型異形肝移植モデルにおける動脈リコネクション後の合併症の減少

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