The authors would like to thank the many individuals who have worked on and perfected this technique and have been responsible for the generation of many manuscripts both in this lab and others. This work was supported by National Institutes of Health Grant EY12707 (PMS) and an unrestricted grant from Research to Prevent Blindness to Department of Ophthalmology.

注：この手順で使用するすべての動物は、ビジョンの研究のための協会と眼科文の眼科と視覚研究における動物の使用のためだけでなく、セントルイス大学で動物の監督委員会によって定めるガイドラインに従って扱われます。 注：すべての手術器具やソリューションは、目の微生物感染を制限するために手術前に滅菌される。これは、動物は、この手順のいくつかの痛みを経験しない間、我々は、鎮痛薬を使用しないことに留意すべきである。すべての鎮痛薬は、抗炎症性であり、角膜移植応答は、炎症を伴うため、系抗炎症薬の使用は、角膜移植不全に関与しているかの要因を決定する我々の能力を危うくするので、この理由による。 1.麻酔<ol style="margin-left: 40px;"><li>ケタミン（86mgのIP注射によって、全身麻酔下で、ドナーとレシピエントマウスを置きます。98ミリグラム/ kg）およびキシラジン（13.04 mg / kgを）。 </li><li>通常、時間に30分を要する作業中は麻酔下でレシピエントマウスにしてください。その結果、常に意識を取り戻すの兆候のためのマウスを監視します。 </li><li>動物は乾燥を防ぐために麻酔をかけた後に手術を受けることはありません目にpuralube軟膏を適用します。 </li></ol> 2。角膜移植<ol style="margin-left: 40px;"><li>ドナー角膜ボタンを取得する。 <ol><li>動物が完全に麻酔したら、1％トロピカミドおよび2.5％塩酸フェニレフリンの点眼剤のカップルの投与により十分な散瞳を達成。 </li><li>水平に頑丈な可動支持体の上に置かれ、ボード上のドナー動物の頭を置きます。目は操作全体を通して水平な位置にあることを保証するために、首全体でテープのストリップで頭を固定します。 </li><li>アウト、先端がメチルブルーで染色した直径2mmのトレフィンを使用して、中央角膜移植部位を裏打ちする。 </li><li>鋭利な刃で、角膜に浸透し、ドナー内皮および基盤となるレンズへの損傷の可能性を減らすためにそれを深めるために前房内にのHealonを注入する。 </li><li> vannasはさみを持つドナー移植片を切除し、使用するまでハンクス平衡塩溶液を含む皿に置く。 </li><li>ドナー移植片が除去された後、CO 2吸入によりドナーマウスを安楽死させる。 </li></ol></li><li>移植床の準備。 <ol><li>受信者に対して2.1.2を通じて、2.1.1で説明したのと同じ手順を繰り返します。 </li><li>直径1.5mmのトレフィンを使用して、受信者の移植部位を概説。 </li><li>鋭利な刃で、角膜に浸透し、下層のレンズへの損傷の可能性を低減することを深めて前房へのHealonを注入する。 </li><li> vannasはさみを使って受信者から概説し、中央の角膜のボタンを外して捨てる。 </li></ol></li><li>縫合移植<ol><li>受取人の角膜のベッドの上で移植ドナーの角膜を置きます。十分なのHealonは、レンズとの直接接触による損傷からドナー内皮細胞を保護するために、ドナー角膜の下にあることを確認してください。 </li><li>超微細な先端マイクロピンセットを使用して、その後オフネクタイ、受信者の側への完全な厚さの90％の深さを持つドナーを通じて、ドナー側に11-0ナイロン縫合糸の最初の一口を置く。 </li><li>角膜が所定の位置に固定されると、角膜は8~10総縫合糸を有し、ドナー角膜がしっかりと整列し、受信者の角膜の移植床に取り付けられるようにmidcardinal結節縫合を行う。 </li></ol></li><li>前房の深化<ol><li>前房内にHBSSまたは気泡を注入することによって前房を深め、静かにセルローススポンジで漏れのため、角膜移植片の完全性をチェック。 注：前房を改質することができない場合、cは確率が高い移植された角膜の将来の評価が非常に困難になり、また、潜在的ドナー角膜内皮機能不全、したがって、移植の失敗につながるataract。 </li></ol></li><li>最終的評価<ol><li>瞳孔が丸いと前房の深さが正常であることを決定するために、目を守ってください。 注：瞳孔が丸いされていない場合、これは、虹彩を縫合中に破損したため、移植片が技術的失敗と見なされていることを示します。 </li><li>目に抗生物質軟膏を適用します。オプション：7-0絹縫合糸で眼瞼を閉じます。 </li><li>彼らは完全に目を覚ましていると、その後個別に手術後2日以上のためにそれらを収容するまで、マウスを観察します。 </li></ol></li></ol> 3.抜糸<ol style="margin-left: 40px;"><li>上記のように、蓋縫合糸が使用されているような場合には、マウスを麻酔し、48時間後に蓋縫合糸を除去する。 </li><li>術後7日目のマウスを麻酔。 suturを削除ES角膜移植を確保。縫合糸が除去され、動物が完全に目覚めた後は、そのケージに戻す。 </li></ol> 4.臨床評価<ol style="margin-left: 40px;"><li> 、白内障（レンズの曇り）、前房出血（前房中の血液）、適切な深さではない前房、あるいは角膜のかなりの不透明度を含む処置合併症の兆候に目を調べます。 「妥協」として、これらの合併症を証明するものを考えてみましょうとCO 2吸入によってそれらを安楽死させる。 <ol><li>無麻酔のマウスのすべての検査を実行します。一方、目のより良いビューを有効にするために目をproptoseできるように、このようにしてマウスを拘束片手でマウスを保持する。観測が完了すると、そのケージに動物を返す。 </li></ol></li><li>治療群に不慣れでは角膜移植の兆候を2〜3回、週に移植した角膜を評価するオブザーバを持っている拒絶反応または角膜移植の失敗。手術用顕微鏡またはこれらの観​​察のための水平細隙灯生体顕微鏡のいずれかを使用します。 <ol><li>以下のようにスケールが定義されている0〜5のスケールを使用して不透明度毎に角膜を評価する。 <ol><li>不透明度の兆候を持っていないもの角膜0~10のスコアを割り当てます。 </li><li>最小限の表面的な不透明性を示すもの角膜にスコア1を割り当てます。 </li><li>マイルドで深い不透明度を表示しますが根底にある瞳と虹彩はまだ識別可能である角膜へ2のスコアを割り当てます。 </li><li>虹彩は、瞳孔の余白を除いて詳細に見ることができない、請求間質不透明度を表示角膜3のスコアを割り当てる。 </li><li>密な間質不透明度を表示しない根本的な構造は閲覧できない場合角膜に4のスコアを割り当てます。 </li><li>完全に覆い隠さ瞳孔と虹彩との完全な不透明度と集中的な間質浮腫を表示角膜に5のスコアを割り当てます。 </li></ol></li><li><html>また、これを達成するために1〜8の評価尺度を使用して、血管浸潤（新血管形成）の程度ごとに角膜を評価する4つの等しい四分円からなるとして角膜を表示​​し、これらの象限のそれぞれの中の血管の量を決定するその象限の広範な血管新生のために0（無血管）から2の範囲で得点。最終的な血管新生スコアを計算するために各象限から個々のスコアを追加します。 </li><li>彼らは5週間までの時点で連続する2つの観察のための3のスコアを持っているかのように急性拒絶された角膜を分類します。 </li><li>その角膜5週目で透明であったマウスを分類しますが&gt; 45日の時点で混濁を開発後期長期角膜同種移植片拒絶を受けたように、2つの連続した​​時点で3のスコアで、移植を投稿してください。移植片生存を分析するために、カプラン - マイヤー生存曲線を使用する。 </li></ol></li></ol>モデル5.操作中左：40px; &quot;&gt; <li>脾臓からの単一細胞の調製。 <ol><li>単一細胞を調製するため、第一のドナーマウスを安楽死させる。その後、脾臓を除去します。 </li><li>セルストレーナーに脾臓を置き、それを3ミリリットルのシリンジからシリンジプランジャを混乱させる。 </li><li>ハンクス平衡塩類溶液10mlに再懸濁細胞を洗ってください。 </li><li>細胞懸濁液10μlを削除し、0.4％トリパンの10μlに青追加し、混ぜる。血球計数器にそれを追加し、中央のグリッド内のセルを数える。チューブ内の細胞数は、細胞数×10 4×2（トリパンブルーで希釈係数）10（チューブ内の容積）×である。 </li></ol></li><li>前房内への注入。 <ol><li>前述のようにマウスを麻酔。 </li><li>解剖顕微鏡を使用して注射を行う。各房内注入のために、10 6 0.005ミリリットルボリューム内の脾臓細胞と33 Gの針を取り付け0.25ミリリットルのマイクロシリンジを使用しています。 注：その他の操作モデルの特定の因子が、同所角膜同種移植手術後の果たし得る役割を決定するためのいずれかのアンタゴニストまたはアゴニストとして作用する試薬で動物を処理することによって行うことができる。 </li></ol></li></ol>

<ol>
	<li>Farooq, A. V., Shukla, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Herpes+simplex+epithelial+and+stromal+keratitis:+an+epidemiologic+update.">Herpes simplex epithelial and stromal keratitis: an epidemiologic update.</a> Surv. Ophthalmol. , 448-462 (2012).</li><li>Gipson, I. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-related+changes+and+diseases+of+the+ocular+surface+and.">Age-related changes and diseases of the ocular surface and.</a> Invest. Opthlamol. Vis. Sci. 54, 48-53 (2013).</li><li>Edwards, 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=Indications+for+corneal+transplantation+in+New+Zealand:+1991-1999.">Indications for corneal transplantation in New Zealand: 1991-1999.</a> Cornea. 21, 152-155 (2002).</li><li>Thompson, R. W., Price, M. O., Bowers, P. J., Price, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+survival+after+penetrating+keratoplasty.">Long-term survival after penetrating keratoplasty.</a> Ophthalmol. 110, 1396-1402 (2003).</li><li>Williams, K. A., Roder, D., Esterman, A., Muehlberg, S. M., Coster, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+predictive+of+corneal+graft+survival.+Report+form+the+Australian+Corneal+Graft+Registry.">Factors predictive of corneal graft survival. Report form the Australian Corneal Graft Registry.</a> Ophthalmology. 99, 403-414 (1992).</li><li>Larkin, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+allograft+rejection.">Corneal allograft rejection.</a> Br. J. Ophthalmol. 78, 649-652 (1994).</li><li>Boisjoly, H. 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=Risk+factors+of+corneal+graft+failure.">Risk factors of corneal graft failure.</a> Ophthalmol. 100, 1728-1735 (1993).</li><li>Sugar, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recipient+Risk+Factors+for+Graft+Failure+in+the+Cornea+Donor+Study.">Recipient Risk Factors for Graft Failure in the Cornea Donor Study.</a> Ophthalmol. 116, 1023-1028 (2009).</li><li>Namba, K., Kitaichi, N., Nishida, T., Taylor, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+regulatory+T+cells+by+the+immunomodulating+cytokines+alpha-melanocyte-stimulating+hormone+and+transforming+growth+factor-beta2.">Induction of regulatory T cells by the immunomodulating cytokines alpha-melanocyte-stimulating hormone and transforming growth factor-beta2.</a> J. Leukoc. Biol. 72, 946-952 (2002).</li><li>Taylor, A. W., Yee, D. G., Streilein, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+nitric+oxide+generated+by+inflammatory+macrophages+by+calcitonin+gene-related+peptide+in+aqueous+humor.">Suppression of nitric oxide generated by inflammatory macrophages by calcitonin gene-related peptide in aqueous humor.</a> Invest. Ophthalmol. Vis. Sci. 39, 1372-1378 (1998).</li><li>Wilbanks, G. A., Mammolenti, M., Streilein, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+on+the+induction+of+anterior+chamber-associated+immune+deviation+(ACAID).+III.+Induction+of+ACAID+depends+upon+intraocular+transforming+growth+factor-beta.">Studies on the induction of anterior chamber-associated immune deviation (ACAID). III. Induction of ACAID depends upon intraocular transforming growth factor-beta.</a> Eur. J. Immunol. 22, 165-173 (1992).</li><li>Volpert, O. V., 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=Inducer-stimulated+Fas+targets+activated+endothelium+for+destruction+by+anti-angiogenic+thrombospondin-1+and+pigment+epithelium-derived+factor.">Inducer-stimulated Fas targets activated endothelium for destruction by anti-angiogenic thrombospondin-1 and pigment epithelium-derived factor.</a> Nat. Med. , 8-349 (2002).</li><li>Apte, R. S., Sinha, D., Mayhew, E., Wistow, G. J., Niederkorn, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cutting+edge:+role+of+macrophage+migration+inhibitory+factor+in+inhibiting+NK+cell+activity+and+preserving+immune+privilege.">Cutting edge: role of macrophage migration inhibitory factor in inhibiting NK cell activity and preserving immune privilege.</a> J. Immunol. 160, 5693-5696 (1998).</li><li>Kennedy, M. C., 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=Novel+production+of+interleukin-1+receptor+antagonist+peptides+in+normal+human+cornea.">Novel production of interleukin-1 receptor antagonist peptides in normal human cornea.</a> J. Clin. Invest. 95, 82-88 (1995).</li><li>Shimmura-Tomita, M., Wang, M., Taniguchi, H., Akiba, H., Yagita, H., Hori, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Galectin-9-mediated+protection+from+allo-specific+T+cells+as+a+mechanism+of+immune+privilege+of+corneal+allografts.">Galectin-9-mediated protection from allo-specific T cells as a mechanism of immune privilege of corneal allografts.</a> PLoS One. 8, (2013).</li><li>Goldberg, M. F., Ferguson, T. A., Pepose, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+cellular+adhesion+molecules+in+inflamed+human+corneas.">Detection of cellular adhesion molecules in inflamed human corneas.</a> Ophthalmol. 101, 161-168 (1994).</li><li>Stuart, P. M., Griffith, T. S., Usui, N., Pepose, J. S., Yu, X., Ferguson, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD95+ligand+(FasL)-induced+apoptosis+is+necessary+for+corneal+allograft+survival.">CD95 ligand (FasL)-induced apoptosis is necessary for corneal allograft survival.</a> J Clin Invest. 99, 396-402 (1997).</li><li>Yamagami, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+Fas-Fas+ligand+interactions+in+the+immunorejection+of+allogeneic+mouse+corneal+transplants.">Role of Fas-Fas ligand interactions in the immunorejection of allogeneic mouse corneal transplants.</a> Transplantation. 64, 1107-1111 (1997).</li><li>Stuart, P. M., Pan, F., Plambeck, S., Ferguson, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fas/Fas+ligand+interactions+regulate+neovascularization+in+the+cornea.">Fas/Fas ligand interactions regulate neovascularization in the cornea.</a> Invest. Ophthalmmol. Vis. Sci. 44, 93-98 (2003).</li><li>Stuart, P. M., Yin, X. T., Pan, F., Haskova, Z., Plambeck, S., Ferguson, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibitors+of+matrix+metalloproteinases+activity+prolong+corneal+allograft+acceptance+by+increasing+FasL+expression.">Inhibitors of matrix metalloproteinases activity prolong corneal allograft acceptance by increasing FasL expression.</a> Invest. Ophthalmol. Vis. Sci. 45, 1169-1173 (2004).</li><li>Joo, C. -. K., Pepose, J. S., Stuart, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T-cell+mediated+responses+in+a+murine+model+of+orthotopic+corneal+transplantation.">T-cell mediated responses in a murine model of orthotopic corneal transplantation.</a> Invest.Ophthalmol. Vis. Sci. 36, 1530-1540 (1995).</li><li>Sonoda, Y., Sano, Y., Ksander, B., Streilein, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+cell-mediated+immune+responses+elicited+by+orthotopic+corneal+allografts+in+mice.">Characterization of cell-mediated immune responses elicited by orthotopic corneal allografts in mice.</a> Invest. Ophthalmol. Vis. Sci. 36, 427-434 (1995).</li><li>Sano, Y., Osawa, H., Sotozono, C., Kinoshita, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytokine+expression+during+orthotopiccorneal+allograft+rejection+in+mice.">Cytokine expression during orthotopiccorneal allograft rejection in mice.</a> Invest. Ophthalmol. Vis. Sci. 39, 1953-1957 (1998).</li><li>Haskova, Z., Usui, N., Ferguson, T. A., Pepose, J. S., Stuart, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD4++T+cells+are+critical+in+corneal+but+not+skin+allograft+rejection.">CD4+ T cells are critical in corneal but not skin allograft rejection.</a> Transplantation. 69, 483-488 (2000).</li><li>Tan, 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=Immunological+disruption+of+antiangiogenic+signals+by+recruited+allospecific+T+cells+leads+to+corneal+allograft+rejection.">Immunological disruption of antiangiogenic signals by recruited allospecific T cells leads to corneal allograft rejection.</a> J. Immunol. 188, 5962-5969 (2012).</li><li>Dana, M. R., Yamada, J., Streilein, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topical+interleukin-1+receptor+antagonist+promotes+corneal+transplant+survival.">Topical interleukin-1 receptor antagonist promotes corneal transplant survival.</a> Transplantation. 63, 1501-1507 (1997).</li><li>Cunnusamy, K., Chen, P. W., Niederkorn, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-17A-dependent+CD4+CD25++regulatory+T+cells+promote+immune+privilege+of+corneal+allografts.">IL-17A-dependent CD4+CD25+ regulatory T cells promote immune privilege of corneal allografts.</a> J. Immunol. 186, 6737-6745 (2011).</li><li>Fu, 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=Arginine+depletion+as+a+mechanism+for+the+immune+privilege+of+corneal+allografts.">Arginine depletion as a mechanism for the immune privilege of corneal allografts.</a> Eur. J. Immunol. 41, 2997-3005 (2011).</li><li>Medina, C. A., Rowe, A. M., Yun, H., Knickelbein, J. E., Lathrop, K. L., Hendricks, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Azithromycin+treatment+increases+survival+of+high-risk+corneal+allotransplants.Cornea.">Azithromycin treatment increases survival of high-risk corneal allotransplants.Cornea.</a> , 32-658 (2013).</li><li>Cho, Y. K., Zhang, X., Uehara, H., Young, J. R., Archer, B., Ambati, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+Endothelial+Growth+Factor+Receptor+1+morpholino+increases+graft+survival+in+a+murine+penetrating+keratoplasty.">Vascular Endothelial Growth Factor Receptor 1 morpholino increases graft survival in a murine penetrating keratoplasty.</a> Invest. Ophthalmol. Vis. Sci. 53, 8458-8471 (2012).</li><li>Kim, H. K., Choi, J. A., Uehara, H., Zhang, X., Ambati, B. K., Cho, Y. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Presurgical+corticosteroid+treatment+improves+corneal+transplant+survival+in+mice.">Presurgical corticosteroid treatment improves corneal transplant survival in mice.</a> Cornea. 32, 1591-1598 (2013).</li><li>Yamazoe, K., Yamazoe, K., Shimazaki-Den, S., Shimazaki, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prognostic+factors+for+corneal+graft+recovery+after+severe+corneal+graft+rejection+following+penetrating+keratoplasty.">Prognostic factors for corneal graft recovery after severe corneal graft rejection following penetrating keratoplasty.</a> BMC Ophthalmol. 13, 5 (2013).</li><li>Panda, A., Vanathi, M., Kumar, A., Dash, Y., Priya, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+graft+rejection.">Corneal graft rejection.</a> Surv. Ophthalmol. 52, 375-396 (2007).</li><li>Patel, S. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Graft+survival+and+endothelial+outcomes+in+the+new+era+of+endothelial+keratoplasty.">Graft survival and endothelial outcomes in the new era of endothelial keratoplasty.</a> Exp. Eye Res. 95, 40-47 (2012).</li><li>Anshu, A., Price, M. O., Tan, D. T., Price, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+keratoplasty:+a+revolution+in+evolution.">Endothelial keratoplasty: a revolution in evolution.</a> Surv. Ophthalmol. 57, 236-252 (2013).</li></ol>

The authors have no competing financial interests.

ここで説明する角膜移植のマウスモデルは、因子が最良の拒絶15,17,18,20、26-30と角膜の受け入れ21-25両方に関連付けられているかを予測するモデルでのヒト角膜同種移植片拒絶を研究する研究者を可能に同種移植片。患者が治療又は予防のいずれか拒絶31のいずれかに局所または全身ステロイド治療を与えられたヒト角膜移植とは異なり、このモデルは、典型的には、?...

角膜移植は、ヒトで行わ移植の最も成功したと一般的なタイプの一つです。この手術が行われる理由は、損傷、感染症1、または非感染性角膜疾患2の他の形態の結果である。アメリカのアイバンク協会からフィギュアは、46000の上には、（：restoresight.org/eye_banks/eye_banks.htmlでのWebサイトを参照してください）​​2011年に行われたことを示している。その成功の指標は、同種移植片、角膜1年間故障率10〜15％の範囲で5年での成功率が70％3-8を超えているということである。多くの研究が示したように、角膜の同種移植の成功は、直接目に免疫学的特権部位であるという事実に関連している。免疫特権部位として角膜の状態を担当する要因は、角膜内の両方血管およびリンパ管の欠如は、抗原提示細胞の相対的欠如、suppres角膜によって産生される因子を含むの免疫エフェクターfuntions 9-15、MHCの低発現は16抗原、およびFasLの17-20の発現。 しかし、成功のためのこれらの移植片を素因これらの要因にもかかわらず、それらは拒絶3-7を受けない。したがって、この拒絶反応を媒介するそれらのメカニズムを理解するだけでなく、拒絶反応を防止するために、様々な治療法をテストする非常に重要である。そのために、我々はここで、制御の実験環境での角膜移植を研究するために20年以上にわたって使用されている角膜移植のマウスモデルを記述する。移植応答が最終的な移植組織が ​​失敗したか成功したかどうかを決定する協調して働いて、さまざまな要因が関与するので、任意のインビトロモデルにおけるこれらの要因の重要性を理解することは不可能である。結果的に、無傷の動物を用いた研究が成功またはfailuどちらにとって重要などのような要因を決定するために必要とされている移植された組織の再。 動物の他の種は角膜移植を研究するために使用されてきたが、他の種を用いて比較した場合、マウスモデルは、いくつかの利点を有する。最初は、特定の遺伝子を発現するかまたは機能移植におけるより良い研究することができる特異的な免疫学的因子の発現を欠くように遺伝子標的化されたマウスの多くの株が存在することである。また、動物の多くの他の種は存在しないマウスに特異的な多くの試薬（因子を中和する組換え因子および抗体の両方）がある。これらの要因が存在するため、このモデルは、急性角膜の同種移植片の応答15、17,18,20 -29に関与し、関連する因子を同定するために広く使用されている。さらに、角膜移植に関与する因子の多くは、他の組織の移植において機能的であることが知られている。

<table border="0" cellpadding="0" cellspacing="0" width="573">
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			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:71px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Zeiss Surgical Microscope</td>
			<td style="width:71px;">Zeiss</td>
			<td style="width:119px;">Rebuilt</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1 mL Syringe</td>
			<td>BD</td>
			<td>305122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3 mL Syringe</td>
			<td>BD</td>
			<td>309657</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">10 mL Syringe</td>
			<td>BD</td>
			<td>309602</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Vannus Scissors</td>
			<td style="width:71px;">Stortz</td>
			<td style="width:119px;">E-3387</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">11-0 Sutures</td>
			<td style="width:71px;">Alcon</td>
			<td style="width:119px;">717939M</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Trephine 2.0mm</td>
			<td style="width:71px;">Katena</td>
			<td style="width:119px;">K 2-7520</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Trephine 1.5 mm</td>
			<td style="width:71px;">Katena</td>
			<td style="width:119px;">K 2-7510</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Tricaine Hydrochloride 0.5%</td>
			<td style="width:71px;">Alcon</td>
			<td style="width:119px;">NDC 0065-0741-12</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">Healon</td>
			<td style="width:71px;">Abbott</td>
			<td style="width:119px;">Healon OVD</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">Forceps</td>
			<td style="width:71px;">FST</td>
			<td style="width:119px;">11251-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">7-0 Sutures</td>
			<td style="width:71px;">Alcon</td>
			<td align="right" style="width:119px;">8065</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">2.5% Phenylephrine HCl</td>
			<td style="width:71px;">Alcon</td>
			<td style="width:119px;">NDC 61314-342-02</td>
			<td></td>
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		<tr height="41">
			<td height="41" style="height:41px;width:216px;">1% Tropicamide</td>
			<td style="width:71px;">Bausch &#38; Lomb</td>
			<td style="width:119px;">NDC-24208-585-59</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hamilton Syringe</td>
			<td style="width:71px;">Hamilton</td>
			<td style="width:119px;">7654-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">33 gauge needle</td>
			<td style="width:71px;">Hamilton</td>
			<td align="right" style="width:119px;">90033</td>
			<td></td>
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		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Cell Strainer (100 &#956;m nylon)</td>
			<td style="width:71px;">BD Falcon</td>
			<td align="right" style="width:119px;">352360</td>
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		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Hemocytometer</td>
			<td style="width:71px;">Cardinal Health</td>
			<td style="width:119px;">B3175</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Trypan Blue</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">T8154</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine corneal transplantation: a model to study the most common form of solid organ transplantation

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each year. While several animal models exist for this procedure and mice are the species that is most commonly used. The reasons for using mice are the relative cost of using this species, the existence of many genetically defined strains that allow for the study of immune responses, and the existence of an extensive array of reagents that can be used to further define responses in this species. This model has been used to define factors in the cornea that are responsible for the relative immune privilege status of this tissue that enables corneal allografts to survive acute rejection in the absence of immunosuppressive therapy. It has also been used to define those factors that are most important in rejection of such allografts. Consequently, much of what we know concerning mechanisms of both corneal allograft acceptance and rejection are due to studies using a murine model of corneal transplantation. In addition to describing a model for acute corneal allograft rejection, we also present for the first time a model of late-term corneal allograft rejection.

角膜移植のマウスモデルが正常角膜同種移植片拒絶19-23と角膜同種移植の受け入れ13、15,16,18、24-27両方のメカニズムを特徴づけるために20年以上にわたって使用されてきた。このモデルは、FasLのを欠く動物は、角膜の同種移植片15を受け入れることができなかったという点で、角膜の同種移植片の受け入れにFasL発現の重要性を確立するために使用した。また、血管内?...

Watch this Scientific Journal Video about Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation at JoVE.com

Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation

Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.

ネズミ角膜移植：固体臓器移植の最も一般的な形態を研究するためのモデル

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each ...

murine-corneal-transplantation-model-to-study-most-common-form-solid

Research

JoVE Journal

Medicine

9.6K ビュー.  Saint Louis University. Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.

ビデオ: Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation

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 Method for Murine Islet Isolation and Subcapsular Kidney Transplantation

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize their blood glucose levels, islet transplantation has been proposed to be a potential treatment for type 1 diabetes 1,2. More recently, advances in human islet transplantation have further strengthened this view 1,3. However, two major limitations prevent islet transplantation from being a widespread clinical reality: (a) the requirement for large numbers of islets per patient, which severely reduces the number of potential recipients, and (b) the need for heavy immunosuppression, which significantly affects the pediatric population of patients due to their vulnerability to long-term immunosuppression. Strategies that can overcome these limitations have the potential to enhance the therapeutic utility of islet transplantation.
Islet transplantation under the mouse kidney capsule is a widely accepted model to investigate various strategies to improve islet transplantation. This experiment requires the isolation of high quality islets and implantation of islets to the diabetic recipients. Both procedures require surgical steps that can be better demonstrated by video than by text. Here, we document the detailed steps for these procedures by both video and written protocol. We also briefly discuss different transplantation models: syngeneic, allogeneic, syngeneic autoimmune, and allogeneic autoimmune.

Transplantation of isolated islets has been proposed to be a potential treatment for type 1 diabetes. Here we describe a method to isolate islets from mouse pancreata and transplant them to the subcapsular space of the kidney.

マウス膵島の単離と被膜下腎移植のための方法

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize ...

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are available to perform mechanistic in vivo studies. While heart transplantation models using a suture technique were first successfully developed in rats, the translation into an equally widespread used murine equivalent was never achieved due the technical complexity of the microsurgical procedure. In contrast, non-suture cuff techniques, also developed initially in rats, were successfully adapted for use in mice1-3. This technique for revascularization involves two major steps I) everting the recipient vessel over a polyethylene cuff; II) pulling the donor vessel over the formerly everted recipient vessel and holding it in place with a circumferential tie. This ensures a continuity of the endothelial layer, short operating time and very high patency rates4.
Using this technique for vascular anastomosis we performed more than 1,000 cervical heart transplants with an overall success rate of 95%. For arterial inflow the common carotid artery and the proximal aortic arch were anastomosed resulting in a retrograde perfusion of the transplanted heart. For venous drainage the pulmonary artery of the graft was anastomosed with the external jugular vein of the recipient5.
Herein, we provide additional details of this technique to supplement the video.

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.
Herein, we provide additional details of this technique to supplement the video.

変更されたカフ法を用いたマウス子宮頸心臓移植モデル

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

異心臓移植のマウスモデルを用いた肺動脈弁の移植

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

Cardiac Catheterization in Mice to Measure the Pressure Volume Relationship: Investigating the Bowditch Effect

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these strategies is to examine their effects on heart function. There are several techniques to measure in vivo cardiac mechanics (e.g., echocardiography, pressure/volume relations, etc.). Compared to echocardiography, real-time left ventricular (LV) pressure/volume analysis via catheterization is more precise and insightful in assessing LV function. Additionally, LV pressure/volume analysis provides the ability to instantaneously record changes during manipulations of contractility (e.g., &#946;-adrenergic stimulation) and pathological insults (e.g., ischemia/reperfusion injury). In addition to the maximum (+dP/dt) and minimum (-dP/dt) rate of pressure change in the LV, an accurate assessment of LV function via several load-independent indexes (e.g., end systolic pressure volume relationship and preload recruitable stroke work) can be attained. Heart rate has a significant effect on LV contractility such that an increase in the heart rate is the primary mechanism to increase cardiac output (i.e., Bowditch effect). Thus, when comparing hemodynamics between experimental groups, it is necessary to have similar heart rates. Furthermore, a hallmark of many cardiomyopathy models is a decrease in contractile reserve (i.e., decreased Bowditch effect). Consequently, vital information can be obtained by determining the effects of increasing heart rate on contractility. Our and others data has demonstrated that the neuronal nitric oxide synthase (NOS1) knockout mouse has decreased contractility. Here we describe the procedure of measuring LV pressure/volume with increasing heart rates using the NOS1 knockout mouse model.

This article describes the measurement of murine left ventricular function via pressure/volume analysis at different heart rates.

ボーディッチ影響を調べる：圧力体積関係を測定するために、マウスにおける心臓カテーテル検査

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these ...

A Porcine Corneal Endothelial Organ Culture Model Using Split Corneal Buttons

Experimental research on corneal endothelial cells is associated with several difficulties. Human donor corneas are scarce and rarely available for experimental investigations as they are normally needed for transplantation. Endothelial cell cultures often do not translate well to in vivo situations. Due to the biostructural characteristics of non-human corneas, stromal swelling during cultivation induces substantial corneal endothelial cell loss, which makes it difficult to perform cultivation for an extended period of time. Deswelling agents such as dextran are used to counteract this response. However, they also cause significant endothelial cell loss. Therefore, an ex vivo organ culture model not requiring deswelling agents was established. Pig eyes from a local slaughterhouse were used to prepare split corneal buttons. After partial corneal trephination, the outer layers of the cornea (epithelium, bowman layer, parts of the stroma) were removed. This significantly reduces corneal endothelial cell loss induced by massive stromal swelling and Descemet&#39;s membrane folding throughout longer cultivation periods and improves general preservation of the endothelial cell layer. Subsequent complete corneal trephination was followed by the removal of the split corneal button from the remaining eye bulb and cultivation. Endothelial cell density was assessed at follow-up times of up to 15 days after preparation (i.e., days 1, 8, 15) using light microscopy. The preparation technique used allows a better preservation of the endothelial cell layer enabled by less stromal tissue swelling, which results in slow and linear decline rates in split corneal buttons comparable to human donor corneas. As this standardized organo-typically cultivated research model for the first time allows a stable cultivation for at least two weeks, it is a valuable alternative to human donor corneas for future investigations of various external factors with regards to their effects on the corneal endothelium.

Here, a step-by-step protocol for the preparation and cultivation of porcine split corneal buttons is presented. As this organo-typically cultivated organ culture model shows cell death rates within 15 days, comparable to human donor corneas, it represents the first model allowing long-term cultivation of non-human corneas without adding toxic dextran.

分割角膜ボタンを用いてのブタ角膜内皮器培養モデル

Experimental research on corneal endothelial cells is associated with several difficulties. Human donor corneas are scarce and rarely available for ...

Development of a Noninvasive, Laser-Assisted Experimental Model of Corneal Endothelial Cell Loss

Nd:YAG lasers have been used to perform noninvasive intraocular surgery, such as capsulotomy for several decades now. The incisive effect relies on the optical breakdown at the laser focus. Acoustic shock waves and cavitation bubbles are generated, causing tissue rupture. Bubble sizes and pressure amplitudes vary with pulse energy and position of the focal point. In this study, enucleated porcine eyes were positioned in front of a commercially available Nd:YAG laser. Variable pulse energies as well as different positions of the focal spots posterior to the cornea were tested. Resulting lesions were evaluated by two-photon microscopy and histology to determine the best parameters for an exclusive detachment of corneal endothelial cells (CEC) with minimum collateral damage. The advantages of this method are the precise ablation of CEC, reduced collateral damage, and above all, the non-contact treatment.

Here, we present a protocol to detach corneal endothelial cells (CEC) from Descemet&#8217;s membrane (DM) using a neodymium:YAG (Nd:YAG) laser as an ex vivo disease model for bullous keratopathy (BK).

角膜内皮細胞損失の非侵襲的レーザー支援実験モデルの開発

Nd:YAG lasers have been used to perform noninvasive intraocular surgery, such as capsulotomy for several decades now. The incisive effect relies on the ...

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

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

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 ...

Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for kidney transplantation (KT). Even though, the use of allografts from extended criteria donors (ECD) could partially ease the shortage of organ donors, ECD organs carry a potentially higher risk for inferior outcomes and postoperative complications. Dynamic organ preservation techniques, modulation of ischemia-reperfusion and preservation injury, and allograft therapies are in the spotlight of scientific interest in an effort to improve allograft utilization and patient outcomes in KT.
Preclinical animal experiments are playing an essential role in translational research, especially in the medical device and drug development. The major advantage of the porcine orthotopic auto-transplantation model over ex vivo or small animal studies lies within the surgical-anatomical and physiological similarities to the clinical setting. This allows the investigation of new therapeutic methods and techniques and ensures a facilitated clinical translation of the findings. This protocol provides a comprehensive and problem-oriented description of the porcine orthotopic kidney auto-transplantation model, using a preservation time of 24 hours and telemetry monitoring. The combination of sophisticated surgical techniques with highly standardized and state-of-the-art methods of anesthesia, animal housing, perioperative follow up, and monitoring ensure the reproducibility and success of this model.

Large animal models play an essential role in preclinical transplantation research. Due to its similarities to the clinical setup, the porcine model of orthotopic kidney auto-transplantation described in this article provides an excellent in vivo setting for the testing of organ preservation techniques and therapeutic interventions.

24時間臓器保存と連続テレメトリを用いた豚モデルにおける異形性腎臓自動移植

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for ...

Intramyocardial Transplantation of MSC-Loading Injectable Hydrogels after Myocardial Infarction in a Murine Model

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of transplanted cells within the injured myocardium, limiting their therapeutic efficacy. Recently, the use of scaffolding biomaterials has gained attention for improving and maximizing stem cell therapy. The objective of this protocol is to introduce a simple and straightforward technique to transplant bone marrow-derived mesenchymal stem cells (MSCs) using injectable hydroxyphenyl propionic acid (GH) hydrogels; the hydrogels are favorable as a cell delivery platform for cardiac tissue engineering applications due to their ability to be cross-linked in situ and high biocompatibility. We present a simple method to fabricate MSC-loading GH hydrogels (MSC/hydrogels) and evaluate their survival and proliferation in three-dimensional (3D) in vitro culture. In addition, we demonstrate a technique for intramyocardial transplantation of MSC/hydrogels in mice, describing a surgical procedure to induce myocardial infarction (MI) via left anterior descending (LAD) coronary artery ligation and subsequent MSC/hydrogels transplantation.

Stem cell-based therapy has emerged as an efficient strategy to repair injured cardiac tissues after myocardial infarction. We provide an optimal in vivo application for stem cell transplantation using gelatin hydrogels that are able to be enzymatically cross-linked.

マウスモデルにおける心筋梗塞後のMSC負荷注入性ヒドロゲルの筋内移植

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of ...