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。ミックス低血糖ダルベッコの変更イーグル培地 (LG DMEM) 10 G/L のウシ血清アルブミン (BSA) と 1 グラム/L のコラゲナーゼ タイプ 1。</li>
	<li>人間の皮下の腹部の脂肪組織を解剖 (n = 8、すべての女性、平均年齢 40 ± 9 歳) 一個 (0.5 cm) の滅菌生物学的 SafetyCabinet の生殖不能外科ブレード サイズ 10、アドソン ブラウン組織鉗子および Metzenbaum はさみを使用してにします。</li>
	<li>切り裂かれたティッシュを滅菌瓶メディア ストレージに格納します。切り裂かれたティッシュの総量の重量を量る、準備された分離培地 (150 ml のボトルあたりの分離媒体の脂肪組織の 50 g) で消化を始めます。プロトコルは、一晩ローラー ミキサーに切り裂かれた脂肪と 4 ° C で分離培地ボトルを格納することによってここで一時停止できます。次の手順を続行する前に 15 分間室温 25 ° C に次の日にボトル、prewarm します。 注: 承認エラスムス MC 医療倫理委員会 (# メック-200) の倫理規定によるとで乳房再建手術を受けるドナーから残り物の材料としてその腹部皮下脂肪の得られた:「適切な二次利用人間の組織」の (< http://www.federa.org>)。残った脂肪はないオプトアウトを二次利用ドナーから使われました。</li>
	<li>シェーカー ・ インキュベータで 150 rpm と 1 h の 37 ° C で滅菌ガラス メディア ストレージ ボトルに切り裂かれた脂肪を消化します。</li>
	<li>50 mL チューブに消化ソリューションを分割し、390 x g で 10 分間のチューブを遠心分離機します。</li>
	<li>培地の準備: 10% 牛胎児血清 (FBS)、50 μ g/mL ゲンタマイシン 1.5 μ G/ml ampothericin B. と LG DMEM</li>
	<li>以下の遠心分離、上澄みを除去し、培養液 (10% 牛胎児血清 (FBS)、50 μ g/mL ゲンタマイシン 1.5 μ G/ml ampothericin B と補われる LG DMEM) 20 mL で細胞ペレットを再懸濁します。390 x g で 5 分間のために再度細胞を遠心し、上清を除去します。</li>
	<li>10 ml の培地 (lg 電子-DMEM 10 %fbs、50 μ g/mL ゲンタマイシン 1.5 μ G/ml ampothericin B) の細胞ペレットを再懸濁し、100 μ m のフィルターを通して細胞懸濁液をフィルターします。</li>
	<li>細胞懸濁液を 50 μ l 添加する (赤血球溶解) の 3% 酢酸/メチレン ブルー溶液 50 μ L を追加、ソリューションをミックス、セルをカウントする 10 μ L を使用します。細胞診断とカウントを実行します。プレート細胞培養培地 (lg 電子-DMEM 30 %fbs、50 μ g/mL ゲンタマイシン 1.5 μ G/ml ampothericin B と) で 40,000 セル/cm2の密度で。</li>
	<li>37 ° c 5% CO2 24 h の湿気のある大気でセルを孵化させなさい。</li>
	<li>アスコルビン酸-2-リン酸 (25 μ g/mL) とひと遺伝子組換え繊維芽細胞成長新鮮なインキュベーション、洗浄暖かい (37 ° C) リン酸の細胞緩衝生理食塩水 (PBS) 細胞の残骸を削除し 10 %fbs 文化メディアとメディアを置き換えると、次追加2 (FGF2、1 ng/mL) を考慮します。</li>
	<li>標準 0.25% トリプシン EDTA 溶液 (T175 フラスコ 3 mL) を使用して 90% 合流でサブカルチャー Asc。3-5 分後、10 ml の培地 (lg 電子-DMEM 10 %fbs、50 μ g/mL ゲンタマイシン 1.5 μ G/ml ampothericin B) の 0.25% トリプシン EDTA 溶液を中和します。 注: の流れ cytometry による一般的な ASC 表面マーカーとトライ系統分化アッセイを以前行ったし、Asc がこの表示プロトコル ASC 特徴21,22を使用してを分離したことを明らかにしました。</li>
	<li>10 ml T175 フラスコ (LG DMEM 添加 10 %fbs、50 μ g/mL ゲンタマイシン 1.5 μ G/ml ampothericin B) 培養液中の細胞を洗うし、遠心分離機 250 x g. 削除で 8 分間細胞の上清 1 ml 培養液中の細胞を再懸濁します。診断とセルをカウントします。</li>
	<li>8,000 セル/cm2の密度で T175 培養フラスコで細胞をプレートします。</li>
	<li>さらに使用する前に培地の 10% ジメチルスルホキシド (DMSO) と液体窒素で残りの Asc を凍結します。</li>
</ol>2. ASC シート準備
<ol><li>FBS、1 mL でプレコート熱応答性培養皿 (直径 3.5 cm)、細胞を播種する前に少なくとも 30 分の 37 ° C でインキュベーター内料理を配置します。</li>
	<li>Asc 標準 0.25% トリプシン EDTA 溶液を用いた 3-5 分 (T175 培養用フラスコの 3 mL) を trypsinize します。Trypsinization 後いずれかの細胞の塊がある場合は、カウントする前に 100 μ m のフィルターを通してセルをフィルターします。</li>
	<li>トリプシン溶液を中和する 10% LG DMEM で FBS (T175 フラスコ 10 mL) と 250 x g に 8 分間遠心する細胞懸濁液上澄みを除去し、10% LG DMEM の 1 mL の細胞を再懸濁します FBS。</li>
	<li>FBS と温度応答性培養皿をコーティングしたインキュベーターから 37 ° c の温暖化のプレートに皿を移動し、お皿から FBS を削除します。</li>
	<li>シード Asc 400,000 セル/cm2 (合計で、3.52 × 106細胞培地皿あたり 2 mL で希釈した)。</li>
	<li>慎重に優しく料理を振ることによって細胞をできるだけ均等に配布します。</li>
	<li>5% CO2と湿気のある大気で 37 ° C で 48 時間培養 ASC シート。 注: は、ASC シート形成を妨げる可能性がありますまたは形成された ASC シートの早期剥離を引き起こす温度低下を防ぐために Asc を播種後インキュベーターのドアを開いて最小化しようと。</li>
</ol>3. 部分的な結腸切除術と大腸吻合
<ol><li>誘導し、雄 Wistar ラット (250-350 g の間の重量を量る) イソフルランの 1 L/分を維持 (1.5-5%)/oxygen 吸入とブプレノルフィン 0.05 mg/kg を筋肉内注入します。</li>
	<li>動物が全身麻酔下で、反射テストを用いた麻酔の深さを評価します。乾燥を防ぐため目にビタミン A 含有の眼軟膏を適用します。麻酔は手術のために十分に考えられている無菌で毛を剃ると、70% エタノールで 2 回外科領域をスプレーで腹部を準備します。滅菌紙カーテン腹部をおいましょう。無菌技術を使用し、全体の手術中に滅菌野を維持します。</li>
	<li>滅菌手術ブレード サイズ 10 で 5 cm の正中線腹部切開を行い Metzenbaum ハサミで切開部を拡張します。識別およびコロンを外在生理食塩水湿らせたガーゼで腹腔内の残りの部分から、コロンをパックします。右、真ん中を特定し、左結腸動脈、腸間膜、ぶっきらぼうホルステッド蚊を使用して各容器の周りを解剖し非吸収性の編みシルク 4/0 各個別容器を縛る。</li>
	<li>結腸盲腸 aborally 1.0 cm と 0.5 cm 尾側腸間膜動脈の間を分離します。Metzenbaum はさみを使用して健康なコロンを縦断面します。結腸切除後 2 つの長い綿棒遠位結腸を通じてトランス肛門し、近位結腸の最後に導入することにより、結腸の近位および遠位端を一緒にもたらします。</li>
	<li>手術顕微鏡を使用して、1 つレイヤー反転 5 縫合 (非吸収性のモノフィラメント ポリアミド 8/0) を使用して縫合で縫合が不十分なエンド ツー エンドの吻合を作成します。コロンの壁のすべての層の縫合を置き、ノット イレウスを配置します。</li>
	<li>ラットを制御または ASC シート グループにランダムに分割します。</li>
</ol>4. ASC シート移植
<ol><li>ASC シートの剥離を容易にする、各種の生体内移植前に 40 分室温に冷却する培養皿を許可します。 注: 剥離後、ASC シートは直径約 2 cm に縮小します。ASC シート生存率は剥離20後少なくとも 3-6 h の高いまま。</li>
	<li>血清 1 ml 無料 LG DMEM 培地を取り外し。</li>
	<li>ゆっくり非外科的鉗子で ASC のシートの縁をつかむと、皿側吻合部のラインの上に ASC シートの。</li>
	<li>慎重に ASC を伸ばしシート約 0.25 cm 非外科的鉗子とコロンのまわりにシートのラップを使用して縫合線の上下です。ラップで、背側 ASC シートまでコロンを持ち上げます。 注: ASC シート付着自発的に吻合部のラインに、組織接着剤を使用する必要があるまたは他の生体材料はありません。コントロール グループは、任意の追加治療を受信しません。</li>
	<li>手袋と楽器を変更したり、綿棒を削除します。腹部にコロンを交換してください。吸収性の縫合糸を使用してを実行している 1 つの層に近い腹部リネア アルバは、ポリグ リコール酸 5/0 を編組。皮下を縫合し、皮膚吸収性の縫合糸を使用してを実行している 1 つの層で一緒に編みポリグ リコール酸 5/0。</li>
</ol>5. 術後評価とフォロー アップ
<ol><li>すぐに 5 mL の温かい生理食塩水の皮下注入と動物を術後水分補給します。体温を維持し、動物が胸骨の横臥を維持するために十分な意識を取り戻すまでバイタル サインを監視するための熱ランプの下で動物を配置します。 注: 手術を受けた動物が他の動物を完全に回復するまでの会社には返されません。回復後、水への無料アクセスと通常ラットの食事を許可します。手術後の痛みを軽減するため 3 日間のすべての 6-8 h 皮下 0.05 mg/kg ブプレノルフィンを管理します。本研究では任意の時に抗生剤を投与しません。</li>
	<li>健康と体重の毎日の臨床評価を取得します。非常に低い健康スコアや重度の体重減少の場合動物は麻酔下にいながら心臓穿刺による急激で安楽死する必要があります。</li>
	<li>術後 3 または日の 7 日目、イソフルランの 1 L/分で再びラットの麻酔 (ブプレノルフィン、なし 1.5-5%)/oxygen 吸入 U 字切開で再手術を行い。腹膜炎、狭窄、線維性の癒着、膿瘍の存在の兆候の腹部をチェックしてください。癒着の重症度をスコアし、腹部、吻合部の地域両方の膿瘍します。</li>
	<li>吻合部を含む、コロンの閉じたセグメント内の空気の吸入による吻合部破裂圧力を決定します。時破裂圧力として最初の空気が漏れる空気の圧力と破裂の場所を記録します。</li>
	<li>各犠牲動物からコロンを削除し、パラフィン埋め込むと 4 μ m 厚いセクションにコロンを切断して次のバッファー 4% ホルムアルデヒド - 大腸吻合を含む - コロンのカットのセグメントを修正します。</li>
	<li>心臓穿刺による急激で吻合セグメントのコレクションの後に直接麻酔中の動物を安楽死させます。呼吸運動、心臓の鼓動の不在と動物の死を確認します。</li>
	<li>ヘマトキシリン ・ エオシンと赤、Picro シリウスなど組織化学的染色を行い、人間のミトコンドリア、ラットの血管内皮細胞 (抗 cd34 陽性) と (CD3 +、CD163 +) 免疫系の細胞に対する抗体で免疫組織化学的染色。</li>
	<li>コンピューター分析のためのスライドをデジタル化し、動物のグループを比較します。</li>
</ol>

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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. 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=Fibrinogen-thrombin+collagen+patch+reinforcement+of+high-risk+colonic+anastomoses+in+rats.">Fibrinogen-thrombin collagen patch reinforcement of high-risk colonic anastomoses in rats.</a> World J Gastrointest Surg. 8 (9), 627-633 (2016).</li><li>Kobayashi, J., 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=Fabrication+of+a+thermoresponsive+cell+culture+dish:+a+key+technology+for+cell+sheet+tissue+engineering.">Fabrication of a thermoresponsive cell culture dish: a key technology for cell sheet tissue engineering.</a> Sci Technol Adv Mat. 11 (1), 014111 (2010).</li><li>Elloumi-Hannachi, I., Yamato, M., 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:+a+unique+nanotechnology+for+scaffold-free+tissue+reconstruction+with+clinical+applications+in+regenerative+medicine.">Cell sheet engineering: a unique nanotechnology for scaffold-free tissue reconstruction with clinical applications in regenerative medicine.</a> J Intern Med. 267 (1), 54-70 (2010).</li><li>Miyahara, Y., 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=Monolayered+mesenchymal+stem+cells+repair+scarred+myocardium+after+myocardial+infarction.">Monolayered mesenchymal stem cells repair scarred myocardium after myocardial infarction.</a> Nat Med. 12 (4), 459-465 (2006).</li><li>Makarevich, P. 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=Enhanced+angiogenesis+in+ischemic+skeletal+muscle+after+transplantation+of+cell+sheets+from+baculovirus-transduced+adipose-derived+stromal+cells+expressing+VEGF165.">Enhanced angiogenesis in ischemic skeletal muscle after transplantation of cell sheets from baculovirus-transduced adipose-derived stromal cells expressing VEGF165.</a> Stem Cell Res Ther. 6, 204 (2015).</li></ol>

著者らは、調査した潜在的な利益相反として解釈される可能性が任意の商業や金融関係の不在を宣言します。

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 ...

transplantation-adipose-tissue-derived-stem-cell-sheet-to-reduce

Research

JoVE Journal

Medicine

7.5K ビュー.  Utrecht University. 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。

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

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

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

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 ...

医学

A Mouse Model of 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 utero transplantation (IUT) of hematopoietic stem cells has been used to successfully treat patients with severe combined immunodeficiency.1,2 In several other conditions, however, IUT has been attempted without success.3 Given these mixed results, the availability of an efficient non-human model to study the biological sequelae of stem cell transplantation and gene therapy is critical to advance this field. We and others have used the mouse model of IUT to study factors affecting successful engraftment of in utero transplanted hematopoietic stem cells in both wild-type mice4-7 and those with genetic diseases.8,9 The fetal environment also offers considerable advantages for the success of in utero gene therapy. For example, the delivery of adenoviral10, adeno-associated viral10, retroviral11, and lentiviral vectors12,13 into the fetus has resulted in the transduction of multiple organs distant from the site of injection with long-term gene expression. in utero gene therapy may therefore be considered as a possible treatment strategy for single gene disorders such as muscular dystrophy or cystic fibrosis. Another potential advantage of IUT is the ability to induce immune tolerance to a specific antigen. As seen in mice with hemophilia, the introduction of Factor IX early in development results in tolerance to this protein.14 
In addition to its use in investigating potential human therapies, the mouse model of IUT can be a powerful tool to study basic questions in developmental and stem cell biology. For example, one can deliver various small molecules to induce or inhibit specific gene expression at defined gestational stages and manipulate developmental pathways. The impact of these alterations can be assessed at various timepoints after the initial transplantation. Furthermore, one can transplant pluripotent or lineage specific progenitor cells into the fetal environment to study stem cell differentiation in a non-irradiated and unperturbed host environment.
The mouse model of IUT has already provided numerous insights within the fields of immunology, and developmental and stem cell biology. In this video-based protocol, we describe a step-by-step approach to performing IUT in mouse fetuses and outline the critical steps and potential pitfalls of this technique.

A Mouse Model of in Utero Transplantation

The mouse model of in utero transplantation is a versatile tool that can be used to study the potential clinical applications of stem cell transplantation and gene therapy in the fetus. In this protocol, we present a general approach to performing this technique

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 protocols are finding their way into clinical practice1,2,3. Getting better results, making the protocols more robust, and finding new sources for implantable cells are the focus of recent research4,5. Investigating the effectiveness of cell therapies is not an easy task and new tools are needed to investigate the mechanisms involved in the treatment process6. We designed an experimental protocol of ischemia/reperfusion in order to allow the observation of cellular connections and even subcellular mechanisms during ischemia/reperfusion injury and after stem cell transplantation and to evaluate the efficacy of cell therapy. H9c2 cardiomyoblast cells were placed onto cell culture plates7,8. Ischemia was simulated with 150 minutes in a glucose free medium with oxygen level below 0.5%. Then, normal media and oxygen levels were reintroduced to simulate reperfusion. After oxygen glucose deprivation, the damaged cells were treated with transplantation of labeled human bone marrow derived mesenchymal stem cells by adding them to the culture. Mesenchymal stem cells are preferred in clinical trials because they are easily accessible with minimal invasive surgery, easily expandable and autologous. After 24 hours of co-cultivation, cells were stained with calcein and ethidium-homodimer to differentiate between live and dead cells. This setup allowed us to investigate the intercellular connections using confocal fluorescent microscopy and to quantify the survival rate of postischemic cells by flow cytometry. Confocal microscopy showed the interactions of the two cell populations such as cell fusion and formation of intercellular nanotubes. Flow cytometry analysis revealed 3 clusters of damaged cells which can be plotted on a graph and analyzed statistically. These populations can be investigated separately and conclusions can be drawn on these data on the effectiveness of the simulated therapeutical approach.

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

We demonstrate how to set up an in vitro ischemia/reperfusion model and how to evaluate the effect of stem cell therapy on postischemic cardiac cells.

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

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

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

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can be used both to investigate the biological responses to injury and to test potential therapies. Contusion or compression injury delivered to the surgically exposed spinal cord are the most widely used models of the pathology. In this report the experimental contusion is performed by using the Infinite Horizon (IH) Impactor device, which allows the creation of a reproducible injury animal model through definition of specific injury parameters. Stem cell transplantation is commonly considered a potentially useful strategy for curing this debilitating condition. Numerous studies have evaluated the effects of transplanting a variety of stem cells. Here we demonstrate an adapted method for spinal cord injury followed by tail vein injection of cells in CD1 mice. In short, we provide procedures for: i) cell labeling with a vital tracer, ii) pre-operative care of mice, iii) execution of a contusive spinal cord injury, and iv) intravenous administration of post mortem neural precursors. This contusion model can be utilized to evaluate the efficacy and safety of stem cell transplantation in a regenerative medicine approach.

Spinal cord injury is a traumatic condition that causes severe morbidity and high mortality. In this work we describe in detail a contusion model of spinal cord injury in mice followed by a transplantation of neural stem cells.

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

Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients with impeded blood flow or with large wounds might be prolonged. Cell-based therapies have appeared as a new technique for the treatment of diabetic ulcers, and cell-sheet engineering has improved the efficacy of cell transplantation. A number of reports have suggested that adipose-derived stem cells (ASCs), a type of mesenchymal stromal cell (MSC), exhibit therapeutic potential due to their relative abundance in adipose tissue and their accessibility for collection when compared to MSCs from other tissues. Therefore, ASCs appear to be a good source of stem cells for therapeutic use. In this study, ASC sheets from the epididymal adipose fat of normal Lewis rats were successfully created using temperature-responsive culture dishes and normal culture medium containing ascorbic acid. The ASC sheets were transplanted into Zucker diabetic fatty (ZDF) rats, a rat model of type 2 diabetes and obesity, that exhibit diminished wound healing. A wound was created on the posterior cranial surface, ASC sheets were transplanted into the wound, and a bilayer artificial skin was used to cover the sheets. ZDF rats that received ASC sheets had better wound healing than ZDF rats without the transplantation of ASC sheets. This approach was limited because ASC sheets are sensitive to dry conditions, requiring the maintenance of a moist wound environment. Therefore, artificial skin was used to cover the ASC sheet to prevent drying. The allogenic transplantation of ASC sheets in combination with artificial skin might also be applicable to other intractable ulcers or burns, such as those observed with peripheral arterial disease and collagen disease, and might be administered to patients who are undernourished or are using steroids. Thus, this treatment might be the first step towards improving the therapeutic options for diabetic wound healing.

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

Adipose-derived stem cells (ASCs) are easily isolated and harvested from the fat of normal rats. ASC sheets can be created using cell-sheet engineering and can be transplanted into Zucker diabetic fatty rats exhibiting full-thickness skin defects with exposed bone and then covered with a bilayer of artificial skin.

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 ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

静脈介在モデル：バイパスグラフト開存を研究するための適切なモデル

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

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

In past years, the cell-sheet construct has spurred wide interest in regenerative medicine, especially for reconstructive surgery procedures. The development of diversified technologies combining adipose tissue-derived stromal cells (ADSCs) with various biomaterials has led to the construction of numerous types of tissue-engineered substitutes, such as bone, cartilage, and adipose tissues from rodent, porcine, or human ADSCs. Extended esophageal endoscopic submucosal dissection (ESD) is responsible for esophageal stricture formation. Stricture prevention remains challenging, with no efficient treatments available. Previous studies reported the effectiveness of mucosal cell-sheet transplantation in a canine model and in humans. ADSCs are attributed anti-inflammatory properties, local immune modulating effects, neovascularization induction, and differentiation abilities into mesenchymal and non-mesenchymal lineages. This original study describes the endoscopic transplantation of an ADSC tissue-engineered construct to prevent esophageal stricture in a swine model. The ADSC construct was composed of two allogenic ADSC sheets layered upon each other on a paper support membrane. The ADSCs were labeled with the PKH67 fluorophore to allow probe-based confocal laser endomicroscopy (pCLE) monitoring. On the day of transplantation, a 5-cm and hemi-circumferential ESD known to induce esophageal stricture was performed. Animals were immediately endoscopically transplanted with 4 ADSC constructs. The complete adhesion of the ADSC constructs was obtained after 10 min of gentle application. Animals were sacrificed on day 28. All animals were successfully transplanted. Transplantation was confirmed on day 3 with a positive pCLE evaluation. Compared to transplanted animals, control animals developed severe strictures, with major fibrotic tissue development, more frequent alimentary trouble, and reduced weight gain. In our model, the transplantation of allogenic ADSCs, organized in double cell sheets, after extended ESD was successful and strongly associated with a lower esophageal stricture rate.

This study reports a successful method of endoscopical adipose tissue-derived stromal cell (ADSC)-sheet transplantation for esophageal stricture prevention after an extended endoscopic submucosal dissection (ESD) in a swine model.

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

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological conditions. Partial hepatectomy also facilitates the increased engraftment and proliferation of transplanted cells by accelerating neovascularization and cell migration towards the liver. Here, we describe a simple protocol for performing 30% hepatectomy and transplantation of cells in the spleen of a non-obese diabetic/severe combined immunodeficient NOD.SCID (NOD.CB17-Prkdcscid/J) mouse. 
In this procedure, two small incisions are made. The first incision is to expose and resect the left lobe of the liver, and another small incision is made to expose the spleen for the intrasplenic transplantation of cells. This procedure does not require any specialized surgical skills, and it can be completed in 5-7 minutes with less stress and pain, faster recovery, and better survival. We have demonstrated the transplantation of hepatocytes isolated from a green fluorescent protein (GFP) expressing mouse (Transgenic C57BL/6-Tg (UBC-GFP) 30Scha/J), as well as hepatocyte like cells of human origin (NeoHep) in partially hepatectomized NOD.SCID mice.

We have described a protocol for performing partial hepatectomy (PHx) and cell transplantation via spleen in NOD.SCID (NOD.CB17-Prkdcscid/J) mice. In this protocol, an incision is made to expose and resect the left lobe of the liver followed by another incision for the intrasplenic transplantation of cells.

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

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. To better understand the mechanism behind these surgical procedures, a rat partial liver transplantation (PLTx) model was established for relevant surgical studies. Because of the complexity of the rat PLTx model, a protocol with detailed descriptions is required. An article published previously reported a protocol in which ex vivo hepatectomy was used to achieve 50% rat PLTx. In contrast to this protocol, we introduced a re-arterialized PLTx with an in vivo 70% hepatectomy. An updated vessel-oriented hepatectomy was incorporated into the rat PLTx to refine the microsurgical procedure. The portal veins and hepatic arteries of the left lateral lobe and the median lobe were individually dissected and ligated before removal of the liver parenchyma, thereby decreasing the probability of bleeding in the remnant liver stump. Furthermore, an end-to-side vessel anastomosis between the common hepatic artery and the enlarged proper hepatic artery was introduced to re-arterialize the hepatic artery. By using this end-to-side vessel anastomosis technique, the diameter of the anastomosis was enlarged, thereby decreasing the difficulty of hand suture and maintaining a high rate of anastomotic patency. Moreover, the cuff anastomosis of the infrahepatic vena cava was slightly modified. A section of circumferential liver parenchyma around the vena cava of a recipient was preserved during cuff anastomosis to maintain the three-dimensional shape of the vascular lumen. This section of liver parenchyma was removed after completing the anastomosis. With this modification, the step involving placement of stay sutures was omitted, thereby further shortening the cuff anastomosis time. By using this protocol of rat PLTx, a low liver enzyme level, an intact liver lobular architecture and a high survival rate were achieved after microsurgery.

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

Here, a protocol involving re-arterialized rat partial liver transplantation is presented. Specifically, 70% liver was resected in vivo by using an updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner. The cuff technique was modified to shorten the anastomosis time of the infrahepatic vena cava.

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. ...