腎臓研究英国、エジンバラの外科ロイヤルカレッジと臓器移植の欧州協会からの資金調達は、この研究を支持した。

適切な国や地方の機関倫理、動物実験を行う前になされるべきである。受信者の外科医は2.1 2.8に出演しながら、具体的に、英国で以下の実験は、ドナー外科医が行うべき2 microsurgeonsが同時に動作するために利用可能である動物（科学的手続）法1986の下で実施された、1.1から1.16、その後3.1から3.5のステップ。一人のオペレータのためのステップを順次追跡することができる。 1ドナーの準備注：ここで紹介する手順は、ドナーC57BL / 6 BM12と受信者の20グラムよりも大きい体重を8から16週齢歳のC57BL / 6雄マウス用です。しかし、この手順は、再現性のマウス系統の多様で行うことができる。代表的な結果のセクションに示したデータは、C57BL / 6、C57BL / 6 BM12及びBALB / cマウスから得た。 <ol><li>行動手順を使う可能な限り無菌の動作領域を維持するための努力とgの無菌手術器具や消耗品（オートクレーブ処理）、。 2外科医が利用可能な場合に、受信者の準備と同時に、ドナーの準備を行います。 </li><li>メデトミジン（0.5ミリグラム/ kg）および塩酸ケタミン（/ kgを200 mg）を腹腔内注射（31G針）でマウスを麻酔。全体の手順のために十分な時間を提供する4時間維持される麻酔薬平面におけるこの結果は、実行される。補足麻酔は必要ありません。 </li><li>マウスは（つま先のピンチへの応答）を麻酔していないことを確認してください。 </li><li>マウスの腹部を剃ると粘着テープを持つ任意の抜け毛を取り除く。 </li><li>無菌ドレープ加熱されたマットの上に仰向けにマウスを置き、ゆるく無菌マスキングテープで手足を固定化する。 </li><li>熱傷のための手順全体を通して、マウスを監視します。可能であれば、非電気の熱源を使用しています。 </li><li>眼の潤滑剤とSAを適用します希薄ヨウ素溶液と腹壁をnitize。 </li><li>腹膜腔に入り、3センチメートルゴシック腹部開創を挿入するために腹部正中切開を行います。 </li><li>しっとり腸や手術領域を維持し、内臓の不必要な乾燥を避けるために温め、生理食塩水を適用します。 </li><li>無菌ドレープでマウスを覆い、大動脈、大静脈と左腎臓を露出させるために、オペレータの左（マウスの右）に腸を動かす。 </li><li>胃にラチェット鉗子を適用し、完全に主要な血管や左腎臓を露出するように上方に引っ張る。そのような肝葉、精嚢、および腸のように、離れて手術領域からの組織を後退させる腹部に無菌の綿棒（2mm×2mmと）を減衰させる梱包してください。 </li><li>周囲の外膜、脂肪とぶっきらぼうに先端の細いピンセットを用いて結合組織を切開することにより腹腔内左副腎から左腎臓を分離します。 NEエリア間鉗子の閉じた先端を置きますedは分離され、ゆっくりと鉗子先端がスペースを解剖するために開くことを可能する。 </li><li>結紮した後、9 / Oナイロンで左副腎静脈と左性腺静脈を分割することにより左腎静脈を分離します。腎静脈に近い縫合糸を配置します。 </li><li>ライゲーションおよび縫合糸を残して膀胱に近い7 / O絹縫合糸で尿管を分割するには、長い終了します。腎臓を採取し、尿管吻合のために必要な時にこれらの長い縫合糸端部が使用される。 </li><li>動員し、完全に腎動脈と静脈に大動脈と大静脈上方および下方を分析するには、ぶっきらぼうに血管の正面と側面からリンパ管や脂肪を切開する先端の細い鉗子を使用しています。 </li><li>大動脈と大静脈の間の組織面を見つけて、ゆっくりと鉗子を広げた。ゆっくりと、最小限の外傷で組織を広げるために細かい傾いて鉗子の先端を開きます。 </li><li>角度の付いた罰金を渡すことで、腎動脈に上位および下位大動脈周囲の緩い7 / Oシルクネクタイ血管の前後を通して縫合糸を引くの周り先端鉗子。これらの縫合糸はウィスコンシン大学液の逆行性灌流を可能にするために、腎臓の取得の前に強化される。 </li></ol> 2受信者の準備ドナーの準備のフォローあたりとしては1.2〜1.8を繰り返します。 <ol><li>大動脈、大静脈を露出させるために、オペレータの右（マウスの左）に腸を移動します。滅菌生理食塩水に浸した綿棒で腸をカバー。 </li><li>右腎動脈を連結することにより、右腎摘出術を行い、7/0絹縫合糸と一緒に静脈、その後分割します。 </li><li> 7 / O絹縫合および除算と右尿管を連結。 </li><li>動員し、ステップ1.14と1.15で説明したように、完全に腎動脈と静脈に劣る大動脈と大静脈を解剖。大動脈と大静脈間の完全な解剖を確認してください。大静脈とaortの前方に動作する内部精巣動脈を保つように注意してくださいリンパ管の束と一緒に。 </li><li> 、大静脈と静脈から脊椎に後腹膜で実行されている腰椎の血管を識別し、連続9 / O絹糸で結紮、血管を分割する必要はありません。 </li><li>血管吻合のための間にスペースを微小血管クランプを配置するために十分なスペースを特定します。 </li><li>背側陰茎静脈を介して静脈内ヘパリン（5単位）を管理します。 </li></ol> 3ドナー腎臓検索動脈循環から腎臓を分離するために劣ると優れた大動脈の周囲に配置された7 / Oシルクネクタイを締めます。 <ol><li>大動脈に針（31G）とウィスコンシン大学溶液0.5ml - 逆行冷たい0.2を注入。 </li><li>縫合糸内の大動脈を分割し、腎臓と大動脈の長さの腎動脈を除去するために大静脈との接合部で腎静脈を分割。大動脈から生じる腰椎血管を分割し、それらが存在する場合、wのithoutライゲーション。 </li><li>培養皿中の滅菌綿棒で冷生理食塩水で腎臓を置きます。 </li><li>頸椎脱臼によってドナーマウスを安楽死させる。 </li></ol> 4。腎臓移植 - 腎臓の準備<ol><li>腎動脈の向かい縦方向に直接大動脈壁を分割することにより、大動脈パッチを作成します。動脈吻合を行う際にライゲートまたは回避する必要があるパッチ内の任意の血管の内腔を特定します。 </li><li>下方上方腎静脈内腔の外側から内側に10 / O縫合糸を配置し、第2の別の縫合糸。副腎静脈および性腺静脈を分割するために使用される縫合糸は、容器を方向付けするために使用することができる。 </li><li>受信者の右脇腹（ 図1.1）に腎臓を置き、血管吻合は綿棒や他の楽器に絡めなるの縫合することなく完了することができることを確認するために、縫合糸が十分に配置されていることを確認してください。 </li></ol> 5。腎臓Transplanテーション - 血管吻合最初に下方その後上方大静脈と大動脈を取り囲む微小血管クランプを適用します。 <ol><li>大静脈の前壁を穿刺することによって針（31G）でventomyを行います。 0.9％NaClを約50μLを注入して大静脈から血液をフラッシュ。 </li><li>長さはドナー腎静脈の直径に相当になるように静脈切開内でそれらを開くことによって、先端の細いピンセットを用いて静脈切開を広げる。 </li><li>静脈切開（ 図1.2）の頂点に最初の（腎静脈の内腔に既にある）10 / O優れた縫合糸を配置し、劣っ頂点に達するまで（ 図を実行している縫合糸での吻合の後壁に参加1.3）。 </li><li> 10 / O劣る縫合糸を配置した後、後壁から実行している縫合糸に結びつの結び目（ 図1.4）で最初に接続します。 </li><li>劣った縫合糸を使用したフロントWALを作成吻合lの連続縫合で（ 図1.5）と優れた頂点（ 図1.6）で、優れた縫合糸端に接続します。 </li><li>鉗子で大動脈壁を拾うとハサミで楕円形のパッチを（ほぼ大動脈切開は大動脈の周囲の長さで三回腎動脈内腔の第五でなければならない）切断することによって大動脈切開を作成します。 </li><li>外部から内部への大動脈パッチの優れた点（ 図1.7）で10 / O縫合糸を配置し、大動脈切開の優れた頂点に血管外に結ぶために大動脈を通過する。 </li><li>受信者の大動脈にドナーパッチを実行している吻合縫合糸が上方に始まると動脈吻合（ 図1.8）を作成します（ 図1.9）きつすぎる縫合糸を結びつけることによって吻合を収縮しないように注意してください。 </li><li> （腎臓を再灌流まず、優れたクランプ劣る血管クランプを取り外し図1.10および1.11）。尿管に十分な血液供給が達成されている場合尿管の蠕動可視多分見。 </li></ol> 6腎臓移植 - 尿管吻合<ol><li>腹壁から膀胱の添付ファイルを分割します。 </li><li>両方の壁を介して膀胱の左から右へ針（21G）を渡します。 </li><li>針管腔内にストレート鉗子先端を置き、鉗子が今膀胱の左側から右から左に通過しているように、膀胱を通って両方がバックに渡す。 </li><li>尿管が右サイドから次に左膀胱不良で通過するように、膀胱を経てドナー尿管上の縫合糸を描画します。 </li><li>ラウンドボディの針で9 / Oナイロン縫合糸のエントリポイントを中心に3つの結節縫合で膀胱の外膜への尿管の外膜を縫合。 </li><li>こうして目を開いて、合字に尿管の近位にカット電子の尿は、尿が流れると尿管の端部が膀胱の体内に引っ込めることができるように許可する。尿の可視生産はペリ尿管の血管からの出血と一緒に観察することができる。 </li><li>シングル中断された9 / O絹縫合糸で右側膀胱不良を閉じます。 </li></ol> 7。回復と術後ケア元の向きで腹部に胃腸内臓を交換して、6 / O吸収性縫合糸で直筋を近似することにより腹壁を閉じます。 <ol><li>金属の皮膚クリップで皮膚に近似している。 </li><li>部分的atipamazole塩酸塩（10μL/ g）を皮下注射による麻酔を逆転。 </li><li>皮下の0.9％NaCl 1mlを皮下ブプレノルフィン塩酸塩（0.05ミリグラム/ kg）を、流体支援注入に鎮痛を管理します。 </li><li> 24〜48時間、28℃で加温キャビネットにマウスを回復します。までの病気のためのマウスを観察します実験終了。 </li><li> 10日に作動ポスト - 金属の皮膚クリップ7を削除します。 </li><li>実験が完了すると、頚椎脱臼によりマウスを安楽死させる。 </li><li>取り外し、組織学的分析のための対側腎臓および同種移植片を収集します。 </li></ol>

<ol>
	<li>Guild, W. R., Harrison, J. H., Merrill, J. P., Murray, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+homotransplantation+of+the+kidney+in+an+identical+twin.">Successful homotransplantation of the kidney in an identical twin.</a> Trans. Am. Clin. Climatol Assoc. 67, 167-173 (1955).</li><li>Wolfe, R. 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=Comparison+of+mortality+in+all+patients+on+dialysis,+patients+on+dialysis+awaiting+transplantation,+and+recipients+of+a+first+cadaveric+transplant.">Comparison of mortality in all patients on dialysis, patients on dialysis awaiting transplantation, and recipients of a first cadaveric transplant.</a> N. Engl. J. Med. 341, 1725-1730 (1999).</li><li>Nankivell, B. J., Alexander, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Rejection of the Kidney Allograft. N. Engl. J. Med. 363, 1451-1462 (2010).</li><li>Tse, G. H., Hughes, J., Marson, L. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systematic+review+of+mouse+kidney+transplantation.">Systematic review of mouse kidney transplantation.</a> Transplant International. 26, 1149-1160 (2013).</li><li>Skoskiewicz, M., Chase, C., Winn, H. J., Russell, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kidney+transplants+between+mice+of+graded+immunogenetic+diversity.+Transplant.">Kidney transplants between mice of graded immunogenetic diversity. Transplant.</a> Proc. 5, 721-725 (1973).</li><li>Zhang, 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=Pattern+of+liver,+kidney,+heart,+and+intestine+allograft+rejection+in+different+mouse+strain+combinations.">Pattern of liver, kidney, heart, and intestine allograft rejection in different mouse strain combinations.</a> Transplantation. 62, 1267-1272 (1996).</li><li>Qi, 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=Depletion+of+cells+of+monocyte+lineage+prevents+loss+of+renal+microvasculature+in+murine+kidney+transplantation.">Depletion of cells of monocyte lineage prevents loss of renal microvasculature in murine kidney transplantation.</a> Transplantation. 86, 1267-1274 (2008).</li><li>Dang, Z., Mackinnon, A., Marson, L. P., Sethi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tubular+atrophy+and+interstitial+fibrosis+after+renal+transplantation+is+dependent+on+galectin-3.">Tubular atrophy and interstitial fibrosis after renal transplantation is dependent on galectin-3.</a> Transplantation. 93, 477-484 (2012).</li><li>Jabs, W. J., 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=Heterogeneity+in+the+Evolution+and+Mechanisms+of+the+Lesions+of+Kidney+Allograft+Rejection+in+Mice.">Heterogeneity in the Evolution and Mechanisms of the Lesions of Kidney Allograft Rejection in Mice.</a> Am. J. Transplant. 3, 1501-1509 (2003).</li><li>Lin, 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=Deficiency+of+C4+from+Donor+or+Recipient+Mouse+Fails+to+Prevent+Renal+Allograft+Rejection.">Deficiency of C4 from Donor or Recipient Mouse Fails to Prevent Renal Allograft Rejection.</a> Am. J. Pathol. 168, 1241-1248 (2006).</li></ol>

著者らは、開示することは何もない。

動脈吻合を実行するために、最もよく説明した方法は、受信者の大動脈にエンド·ツー·サイド方式では、継続中の腎動脈に、ドナーの遠位大動脈を使用することです。私たちは、より便利であると私たちは信じている人間の腎臓移植​​で行われる「カレルパッチ」ミラーリングと同様、大動脈パッチの使用を記載している。ドナーとレシピエント手術時間の文献の報告が疎であるが、私た...

腎不全の治療のために成功した腎移植は、まずそれ以来、人生2の長さと品質の両方で改善を提供し、世界中の末期腎不全患者のための革新的な治療法となっており、1一卵性双生児の間で1955年に記載された。しかし長期的な移植片の生存は、慢性同種移植片損傷3をもたらし病理学的プロセスの多数によって妨げられてきた。 ヒトでの移植された腎臓の拒絶はimmunosupporessiveレジメンの大幅な改善にもかかわらず、罹患率の主要な原因である。腎移植のマウスモデルを開発の目的は密接にヒト腎移植4で見つかったプロセスおよび病理を複製することである。 Skoskiewicz らは、最初に1973年5において腎移植のマウスモデルを説明した。高度な顕微スキルが必要とされているが、それは価値のあるtといくつかの理由からOOL：マウスゲノムは、十分に特徴付けられており、実験方法とマウスの研究のために利用可能な技術の多種多様があります。 腎移植のマウスモデルを使用して多くのグループがしかし、実験4の期間中、 その場に放置されている他の研究でレシピエントマウスのネイティブ腎臓の私達の記載された方法論1で、生命維持器官などの移植された腎臓を使用している。利点は、マウスが、それによって、マウスへの罹患率と第二の手順による死亡のリスクを低減シングル麻酔して、操作を受けるということです。さらにマウスが緩やかな腎不全の悪影響を受けない。 同種異系拒絶のモデルは、心臓、皮膚などの他の臓器に存在するが、これらは常に腎移植とは直接関係ありません。これらのモデルは、異なるモードとdyを誘発するという証拠がある拒絶反応のナミックスは、例えば、心臓同種移植、腎同種移植における拒絶反応の時間的経過は、特定の菌株の組み合わせ6で大幅に異なっている。私たちは、非トランスジェニックFVB / NJマウスにBALB / c系ドナーにおいて急性腎同種移植片拒絶反応パターンを記載して、このモデルは、T細胞およびマクロファージ7の蓄積による細胞媒介性傷害を示した。代替的に、私たちはまた、間質性線維症および尿細管萎縮を示し、慢性の同種移植片損傷のモデルを記載したこれらのマウスは、単一のMHCクラスII遺伝子座のミスによって特徴付けられるように、これは、C57BL / 6レシピエントにC57BL / 6 BM12ドナーからの腎臓移植から生じる-match 8。 移植の複数の態様は、急性拒絶反応、細胞性および体液性拒絶反応、虚血再灌流障害、および新規治療薬を試用を含む腎移植のマウスモデルを用いて研究されている。 私たちは、外科トンを変更した運転時間を短縮し、手術の容易性を向上させるechnique。特に、私たちは、同時ドナーとレシピエントの準備および連続大動脈パッチ吻合を利用することによって単純化された血管吻合技術を記載している。このビデオと原稿が、この技術の確立を支援するために重要なポイントを提供します。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Surgical Instruments</td>
		</tr>
		<tr>
			<td>Blunt Dissecting Scissors</td>
			<td>Fine Science Tools</td>
			<td>&#160;14072-10</td>
			<td>For skin cutting</td>
		</tr>
		<tr>
			<td>Curved Castoviejo scissors</td>
			<td>Fine Science Tools</td>
			<td>15017-10</td>
			<td>For tissue cutting</td>
		</tr>
		<tr>
			<td>Spring Scissors &#8211; straight</td>
			<td>Fine Science Tools</td>
			<td>15000-08</td>
			<td>For suture cutting</td>
		</tr>
		<tr>
			<td>Toothed forceps 1x2 teeth</td>
			<td>Fine Science Tools</td>
			<td>11021-12</td>
			<td></td>
		</tr>
		<tr>
			<td>2 x Fine Tip forceps (Dumont No.5)</td>
			<td>Fine Science Tools</td>
			<td>11251-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Angled Fine Tip forceps (Dumont No. 5/45)</td>
			<td>Fine Science Tools</td>
			<td>11253-25</td>
			<td>For blunt dissecting</td>
		</tr>
		<tr>
			<td>Curved Fine Tip forcep (Dumont No.7)</td>
			<td>Fine Science Tools</td>
			<td>11273-22</td>
			<td>Useful to pass around vessels</td>
		</tr>
		<tr>
			<td>Curved Crile Haemostat</td>
			<td>Fine Science Tools</td>
			<td>1300-04</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro clip applicator with lock</td>
			<td>Fine Science Tools</td>
			<td>18056-14</td>
			<td></td>
		</tr>
		<tr>
			<td>2 x Micro serrefines spring width 2mm, jaw length 4mm</td>
			<td>Fine Science Tools</td>
			<td>18055-04</td>
			<td>Microvascular clamps</td>
		</tr>
		<tr>
			<td>2 x Colibri 3cm wire retractor</td>
			<td>Fine Science Tools</td>
			<td>17000-03</td>
			<td></td>
		</tr>
		<tr>
			<td>Castroviejo needle holder with lock</td>
			<td>Fine Science Tools</td>
			<td>120660-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Wound clip applicator</td>
			<td>Fine Science Tools</td>
			<td>12031-07</td>
			<td></td>
		</tr>
		<tr>
			<td>7mm wound clips</td>
			<td>Fine Science Tools</td>
			<td>12032-07</td>
			<td>Remove 7 to 10 days after surgery</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Equipment</td>
		</tr>
		<tr>
			<td>OPMI pico microscope</td>
			<td>Carl Zeiss</td>
			<td>S100</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermal cautery unit with fine tip</td>
			<td>Geiger</td>
			<td>150A</td>
			<td></td>
		</tr>
		<tr>
			<td>Heat electronic pad</td>
			<td>Cozee Cumfort</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Euroklav 23-S</td>
			<td>Melag</td>
			<td>n/a</td>
			<td>Autoclave</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Disposable equipment</td>
		</tr>
		<tr>
			<td>7/O Silk braided suture</td>
			<td>Pearsall</td>
			<td>30514</td>
			<td></td>
		</tr>
		<tr>
			<td>10/O Dafilon (polyamide) suture</td>
			<td>B-Braun</td>
			<td>&#160;G1118099</td>
			<td></td>
		</tr>
		<tr>
			<td>6/O Vicryl (plygalectin)</td>
			<td>Ethicon</td>
			<td>W9537</td>
			<td></td>
		</tr>
		<tr>
			<td>Regular bevel needle, 1 inch, 21G</td>
			<td>Bection, Dickinson and Company</td>
			<td>305175</td>
			<td>For ureteric anastamosis</td>
		</tr>
		<tr>
			<td>Regular bevel needle, 5/8 inch, 25G</td>
			<td>Bection, Dickinson and Company</td>
			<td>305122</td>
			<td></td>
		</tr>
		<tr>
			<td>Regular bevel needle, 1/2 inch, 30G</td>
			<td>Bection, Dickinson and Company</td>
			<td>304000</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin needle 1ml, 29G</td>
			<td>Bection, Dickinson and Company</td>
			<td>324827</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin needle 0.3ml, 30G</td>
			<td>Bection, Dickinson and Company</td>
			<td>324826</td>
			<td></td>
		</tr>
		<tr>
			<td>1 ml syringe slip tip</td>
			<td>Bection, Dickinson and Company</td>
			<td>300184</td>
			<td></td>
		</tr>
		<tr>
			<td>5 ml syringe slip tip</td>
			<td>Bection, Dickinson and Company</td>
			<td>302187</td>
			<td></td>
		</tr>
		<tr>
			<td>Wypall paper swabs</td>
			<td>Kimberley-Clark</td>
			<td>L40</td>
			<td>sterilised by autoclave</td>
		</tr>
		<tr>
			<td>Cotton wool buds</td>
			<td>Johnson and Johnson</td>
			<td>n/a</td>
			<td>sterilised by autoclave</td>
		</tr>
		<tr>
			<td>Plain drapes</td>
			<td>Guardian</td>
			<td>CB03</td>
			<td>sterilised by autoclave</td>
		</tr>
		<tr>
			<td>Cell culture dish 60mm x 15mm</td>
			<td>Corning Incorporated</td>
			<td>430166</td>
			<td></td>
		</tr>
		<tr>
			<td>Dispensing Pin</td>
			<td>B-Braun</td>
			<td>DP3500L / 413501</td>
			<td>Used with NaCl 0.9%</td>
		</tr>
		<tr>
			<td>Re-agents and Drugs</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>(Lacri-Lube) White soft paraffin 57.3%, mineral oil 42.5% and lanolin alcohols 0.2%</td>
			<td>Allergan Ltd</td>
			<td>21956GB10X</td>
			<td></td>
		</tr>
		<tr>
			<td>(Videne) Povidone-iodine 10%</td>
			<td>Ecolab Ltd</td>
			<td>PL 04509/0041</td>
			<td></td>
		</tr>
		<tr>
			<td>(Vetalar V) Ketamine hydrochloride</td>
			<td>Pfizer Animal Health</td>
			<td>Vm 42058/4165</td>
			<td>100mg/ml solution (dose 200mg/kg)</td>
		</tr>
		<tr>
			<td>(Domitor) Medetomidine hydrochloride&#160;</td>
			<td>Orion Pharma</td>
			<td>Vm 06043/4003</td>
			<td>1mg/ml (dose 0.5mg/kg)</td>
		</tr>
		<tr>
			<td>(Vetergesic) Bupernorphine hydrochloride&#160;</td>
			<td>Alsto Animal Health</td>
			<td>Vm 00063/4002</td>
			<td>0.3mg/ml (dose 0.05mg/kg)</td>
		</tr>
		<tr>
			<td>(Antisedan) Atipamezole hydrochoride</td>
			<td>Orion Pharma</td>
			<td>Vm 06043/4004</td>
			<td>5mg/ml (dose 2mg/kg)</td>
		</tr>
		<tr>
			<td>University of Wisconsin Solution</td>
			<td>Belzer Bridge to Life</td>
			<td>n/a</td>
			<td>dose approximately 500 microlitres/mouse</td>
		</tr>
		<tr>
			<td>NaCl 0.9%</td>
			<td>Baxter</td>
			<td>FKE1323</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin Sulphate</td>
			<td>non-proprietary</td>
			<td>n/a</td>
			<td>5000units/ml (dose 5units/mouse)</td>
		</tr>
	</tbody>
</table>

mouse kidney transplantation: models of allograft rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

腎臓の同種移植片拒絶は、移植された腎臓（ 図2）のmethacarn固定パラフィン包埋組織切片の組織学的分析によって評価することができる。腎臓の虚血再灌流障害における同系マウスの結果の間に腎臓の同系移植移植は、しかし、4週間までに細管を回収し、ネイティブの腎臓への組織学的に同等であるしている。急性拒絶反応は、BALB / cレシピエントにC57BL / 6腎臓移植によってモデ...

Watch this Scientific Journal Video about Mouse Kidney Transplantation: Models of Allograft Rejection at JoVE.com

Mouse Kidney Transplantation: Models of Allograft Rejection

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

マウス腎臓移植：同種移植片拒絶のモデル

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

mouse-kidney-transplantation-models-of-allograft-rejection

Research

JoVE Journal

Medicine

20.4K ビュー.  The University of Edinburgh. Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

ビデオ: Mouse Kidney Transplantation: Models of Allograft Rejection

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 ...

医学

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

使用したラット腎移植後の急性拒絶反応の非侵襲的イメージング<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis

Vascular rejection that leads to transplant arteriosclerosis (TA) is the leading representation of chronic heart transplant failure. In TA, the immune system of the recipient causes damage of the arterial wall and dysfunction of endothelial cells and smooth muscle cells. This triggers a pathological repair response that is characterized by intimal thickening and luminal occlusion. Understanding the mechanisms by which the immune system causes vasculature rejection and TA may inform the development of novel ways to manage graft failure. Here, we describe a mouse aortic interposition model that can be used to study the pathogenic mechanisms of vascular rejection and TA. The model involves grafting of an aortic segment from a donor animal into an allogeneic recipient. Rejection of the artery segment involves alloimmune reactions and results in arterial changes that resemble vascular rejection. The basic technical approach we describe can be used with different mouse strains and targeted interventions to answer specific questions related to vascular rejection and TA.

We describe a protocol for aortic interposition grafting in mice. The goal of the protocol is to provide a model with which to study pathological processes and therapeutic strategies relevant to alloimmune reactions in arteries and the resultant arterial changes that contribute to organ transplant failure.

同種免疫により誘導される血管拒絶反応および移植動脈硬化症のマウスモデル

Vascular rejection that leads to transplant arteriosclerosis (TA) is the leading representation of chronic heart transplant failure. In TA, the immune ...

Murine Kidney Transplant Technique

The first mouse kidney transplant technique was published in 19731 by the Russell laboratory. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique using the donor aorta to form the arterial anastomosis instead of the renal artery was developed and reported in 1993 by Kalina and Mottram 2 with a further advancement coming from the same laboratory in 1999 3. While one can become proficient in this model, a search of the literature reveals that many labs still experience a high proportion of graft loss due to arterial thrombosis. We describe here a technique that was devised in our laboratory that vastly reduces the arterial thrombus reported by others 4,5. This is achieved by forming a heel-and-toe cuff of the donor infra-renal aorta that facilitates a larger anastomosis and straighter blood flow into the kidney.

The goal of this manuscript is to describe the steps required to perform a kidney transplant in a mouse, paying particular attention to the details of the arterial anastomosis.

マウスの腎臓移植​​技術

The first mouse kidney transplant technique was published in 19731 by the Russell laboratory. Although it took some years for other labs to become ...

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a serious problem. Therefore, the acceptance criteria for organ donors have been extended leading to the usage of marginal kidney grafts. These marginal organs tolerate cold storage poorly resulting in increased preservation injury and higher rates of delayed graft function. To overcome the limitations of cold storage, extensive research is focused on alternative normothermic preservation methods.
Ex vivo normothermic organ perfusion is an innovative preservation technique. The first experimental and clinical trials for ex vivo lung, liver, and kidney perfusions demonstrated favorable outcomes.
In addition to the reduction of cold ischemic injury, the method of normothermic kidney storage offers the opportunity for organ assessment and repair. This manuscript provides information about kidney retrieval, organ preservation techniques, and isolated ex vivo normothermic kidney perfusion (NEVKP) in a porcine model. Surgical techniques, set up for the perfusion solution and the circuit, potential assessment options, and representative results are demonstrated.

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

The severe organ shortage has resulted in increased use of marginal kidney grafts for transplantation. This has triggered interest in alternative storage methods, since marginal grafts especially tolerate cold storage poorly. The technique of normothermic ex vivo kidney perfusion (NEVKP) represents a novel preservation method for kidney grafts prior to &#160;transplantation.

正常温度の<em&gt;エクスビボ</em&gt;腎臓の保全のための腎臓の灌流は移植前の移植片

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a ...

Assessment of Kidney Function in Mouse Models of Glomerular Disease

The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function in diabetic nephropathy and podocyte-specific VEGF-A knock-out mice; therefore, this protocol describes the full kidney work-up used in our lab to assess these mouse models of glomerular disease, enabling a vast amount of information regarding kidney and glomerular function to be obtained from a single mouse. In comparison to alternative methods presented in the literature to assess glomerular function, the use of the method outlined in this paper enables the glomerular phenotype to be fully evaluated from multiple aspects. By using this method, the researcher can determine the kidney phenotype of the model and assess the mechanism as to why the phenotype develops. This vital information on the mechanism of disease is required when examining potential therapeutic avenues in these models. The methods allow for detailed functional assessment of the glomerular filtration barrier through measurement of the urinary albumin creatinine ratio and individual glomerular water permeability, as well as both structural and ultra-structural examination using the Periodic Acid Schiff stain and electron microscopy. Furthermore, analysis of the genes dysregulated at the mRNA and protein level enables mechanistic analysis of glomerular function. This protocol outlines the generic but adaptable methods that can be applied to all mouse models of glomerular disease.

This protocol describes a full kidney work-up that should be carried out in mouse models of glomerular disease. The methods allow for detailed functional, structural, and mechanistic analysis of glomerular function, which can be applied to all mouse models of glomerular disease.

糸球体疾患のマウス ・ モデルにおける腎機能の評価

The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function ...

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of renal replacement therapy. Over the past few decades, the rat kidney transplantation model has been used to study the immunological phenomena of rejection and tolerance. This model has become an indispensable tool to test new immunomodulatory pharmaceuticals and regimens prior to proceeding with expensive preclinical large animal studies.
This protocol provides a detailed overview of how to reliably perform orthotopic kidney transplantation in rats. This protocol includes three distinctive steps that increase the probability of success: perfusion of the donor kidney by flushing through the portal vein and the use of a cuff system to anastomose the renal veins and ureters, thereby decreasing cold and warm ischemia times. Using this technique, we have achieved survival rates beyond 6 months with normal serum creatinine in animals with syngeneic or tolerant kidney transplants. Depending on the aim of the study, this model can be modified by pre- or posttransplant treatments to study the acute, chronic, cellular, or antibody-mediated rejection. It is a reproducible, reliable, and cost-effective animal model to study different aspects of kidney transplantation.

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

Orthotopic ラット腎移植: 新規かつ簡便な外科的アプローチ

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of ...

Optical Clearing and Imaging of Immunolabeled Kidney Tissue

Optical clearing techniques render tissue transparent by equilibrating the refractive index throughout a sample for subsequent three-dimensional (3-D) imaging. They have received great attention in all research areas for the potential to analyze microscopic multicellular structures that extend over macroscopic distances. Given that kidney tubules, vasculature, nerves, and glomeruli extend in many directions, which have been only partially captured by traditional two-dimensional techniques so far, tissue clearing also opened up many new areas of kidney research. The list of optical clearing methods is rapidly growing, but it remains difficult for beginners in this field to choose the best method for a given research question. Provided here is a simple method that combines antibody labeling of thick mouse kidney slices; optical clearing with cheap, non-toxic and ready-to-use chemical ethyl cinnamate; and confocal imaging. This protocol describes how to perfuse kidneys and use an antigen-retrieval step to increase antibody- binding without requiring specialized equipment. Its application is presented in imaging different multicellular structures within the kidney, and how to troubleshoot poor antibody penetration into tissue is addressed. We also discuss the potential difficulties of imaging endogenous fluorophores and acquiring very large samples and how to overcome them. This simple protocol provides an easy-to-setup and comprehensive tool to study tissue in three dimensions.

The combination of antibody labeling, optical clearing, and advanced light microscopy allows three-dimensional analysis of complete structures or organs. Described here is a simple method to combine immunolabeling of thick kidney slices, optical clearing with ethyl cinnamate, and confocal imaging that enables visualization and quantification of three-dimensional kidney structures.

免疫標識腎組織の光学クリアリングとイメージング

Optical clearing techniques render tissue transparent by equilibrating the refractive index throughout a sample for subsequent three-dimensional (3-D) ...

Digital Home-Monitoring of Patients after Kidney Transplantation: The MACCS Platform

The MACCS (Medical Assistant for Chronic Care Service) platform enables secure sharing of key medical information between patients after kidney transplantation and physicians. Patients provide information such as vital signs, well-being, and medication intake via smartphone apps. The information is transferred directly into a database and electronic health record at the kidney transplant center, which is used for routine patient care and research. Physicians can send an updated medication plan and laboratory data directly to the patient app via this secure platform. Other features of the app are medical messages and video consultations. Consequently, the patient is better-informed, and self-management is facilitated. In addition, the transplant center and the patient's local nephrologist automatically exchange notes, medical reports, laboratory values, and medication data via the platform. A telemedicine team reviews all incoming data on a dashboard and takes action, if necessary. Tools to identify patients at risk for complications are under development. The platform exchanges data via a standardized secure interface (Health Level 7 (HL7), Fast Healthcare Interoperability Resources (FHIR)). The standardized data exchange based on HL7 FHIR guarantees interoperability with other eHealth solutions and allows rapid scalability to other chronic diseases. The underlying data protection concept is in concordance with the latest European General Data Protection Regulation. Enrollment started in February 2020, and 131 kidney transplant recipients are actively participating as of July 2020. Two large German health insurance companies are currently funding the telemedicine services of the project. The deployment for other chronic kidney diseases and solid organ transplant recipients is planned. In conclusion, the platform is designed to enable home monitoring and automatic data exchange, empower patients, reduce hospitalizations, and improve adherence, and outcomes after kidney transplantation.

The MACCS platform&#160;is a comprehensive telemedicine concept aiming at better outcomes after kidney transplantation by sharing key medical information between patients and physicians. A telemedicine team reviews incoming data to detect potential complications and to improve adherence in kidney transplant recipients to achieve better long-term outcomes.

腎臓移植後の患者のデジタルホームモニタリング:MACCSプラットフォーム

The MACCS (Medical Assistant for Chronic Care Service) platform enables secure sharing of key medical information between patients after kidney ...

Robot-Assisted Kidney Transplantation

This paper describes robot-assisted kidney transplantation (RAKT) from a living donor. The robot is docked between the parted legs of the patient, placed in the supine Trendelenburg position. Kidney allografts are provided by a living donor. Before vascular anastomosis, the kidney allograft is prepared by inserting a double-J stent in the ureter, and the temperature for the anastomosis is lowered by wrapping it in an ice-packed gauze. A 12 mm or 8 mm port for the robotic camera and three 8 mm ports for robotic arms are placed. A peritoneal pouch is created for the kidney allograft by raising the peritoneal flaps on both sides over the psoas muscle before dissecting the iliac vessels and bladder. A 6 cm Pfannenstiel incision is made to insert the kidney into the peritoneal pouch, lateral to the right iliac vessels. 
After clamping the external iliac vein with Bulldogs clamps, a venotomy is performed, and the graft renal vein is anastomosed to the external iliac vein in an end-to-side continuous manner with a 6/0 polytetrafluoroethylene suture. After clamping the graft renal vein, the iliac vein is declamped. This is followed by clamping of the external iliac artery, arteriotomy, arterial anastomosis with a 6/0 polytetrafluoroethylene suture, clamping of the graft renal artery, and declamping of the external iliac artery. Reperfusion is then carried out, and ureteroneocystostomy is performed using the Lich-Gregoir technique. The peritoneum is closed at a few locations with polymer locking clips, and a closed-suction drain is placed through one of the working ports. After deflating the pneumoperitoneum, all incisions are closed.

This paper provides technical details for robot-assisted kidney transplantation from a living donor.

ロボット支援腎移植

This paper describes robot-assisted kidney transplantation (RAKT) from a living donor. The robot is docked between the parted legs of the patient, placed ...