Böbrek Araştırma İngiltere&#39;den finansman, Edinburgh ve Organ Nakli European Society of Surgeons Royal College of bu çalışmayı destekledi.

Uygun ulusal ve yerel kurumsal etik hayvan deneyleri gerçekleştirmeden önce bir yerde olmalıdır. Alıcı cerrah 2,1-2,8 gerçekleştirir iken Özellikle Birleşik Krallık&#39;ta şu deneyleri donör cerrah yapmalıdır iki mikrocerrah aynı anda çalıştırmak için kullanılabilir Hayvanlar (Bilimsel Prosedürleri) Yasası 1986 altında yapılmıştır, 1,1-1,16 sonra 3,1-3,5 adımları . Tek bir operatör için adımlar sırayla takip edilebilir. 1. Donör Hazırlığı Not: Burada sunulan prosedür, verici, C57BL / 6 BM12 ve alıcı 20 g veya daha büyük bir vücut ağırlığı olan 8-16 haftalık yaş arası, C57BL / 6 erkek farelere içindir. Bununla birlikte, bu prosedür, yeniden üretilebilir bir fare suşlarının çeşitli gerçekleştirilebilir. Örnek sonuçlar bölümünde sunulan veriler, C57BL / 6, C57BL / 6 BM12 ve BALB / c farelerinden elde edilmiştir. <ol><li> Davranış prosedürleri istimaluğraşlarla g steril cerrahi aletler ve malzemeler (otoklav), mümkün olduğunca steril çalışma alanını tutmak. İki cerrah olup olmadığını alıcı hazırlanması ile eşzamanlı olarak donör hazırlık yapın. </li><li> Medetomidin (0.5 mg / kg) ve ketamin hidroklorür (/ Kg 200 mg), intraperitoneal enjeksiyon (31G iğne) ile fare anestezisi. Tüm işlemi için yeterli zaman sağlayarak, 4 saat boyunca muhafaza edilen bir anestezik düzlemde Bu sonuç, gerçekleştirilmelidir. Ek anestezi gerekli değildir. </li><li> Fare (ayak tutam hiçbir yanıt) anestezi doğrulayın. </li><li> Farenizin karın Tıraş ve yapışkan bant ile herhangi bir gevşek saç kaldırmak. </li><li> Bir steril drape ısıtılmış mat sırtında fare yerleştirin ve gevşek steril maskeleme bandı ile bacaklarda hareketsiz. </li><li> Termal yanıklar için işlem boyunca fare izleyin. Mümkünse olmayan bir elektrikli ısı kaynağı kullanın. </li><li> Bir göz yağ ve sa UygulaBir iyot çözeltisi ile seyreltilmiş karın duvarı nitize. </li><li> Periton boşluğuna girin ve 3 cm Calibri karın ekartörünü eklemek için karın orta hat kesi olun. </li><li> Nemli bağırsakları ve cerrahi alanda tutmak ve organların gereksiz kurumayı önlemek için ısıtılmış serum fizyolojik uygulayın. </li><li> Steril örtüler ile fare Kapak ve aorta, vena kava ve sol böbrek maruz operatörün sol (Farenizin sağ) için bağırsakları hareket ettirin. </li><li> Mide bir kastanyollanmıştır forcep uygulayın ve tamamen büyük damarları ve sol böbrek maruz superiorda çekin. Uzak gibi karaciğer lobları, seminal kesecikler ve bağırsak gibi cerrahi alanında, dokuların geri çekmek için karın içine steril bezlerden (2 mm x 2 mm) nemli paketleyin. </li><li> Çevre adventia, yağ ve açık açık, ince uçlu forseps kullanılarak bağ dokusu diseksiyon ile periton boşluğuna sol adrenal bezden sol böbrek izole eder. Ne sebeple alanlar arasındaki forseps kapalı ipuçları yerleştirined ayrılır ve yavaş yavaş forcep ipuçları alanı incelemek için açmaya imkan vermek için. </li><li> Bağlanması ve daha sonra 9 / Ç naylon adrenal ven ve sol gonadal ven sol bölerek sol renal ven izole. Renal ven yakın dikiş yerleştirin. </li><li> Ligatı ve dikiş bırakarak mesaneye yakın 7 / Ç ipek sütür ile üreteri bölmek uzun biter. Böbrek toplandı ve üreter anastomoz için gerekli olan zaman, bu uzun ameliyat dikiş ipliği ucu kullanılacaktır. </li><li> Harekete ve tam renal arter superiorda ve alt aort ve vena kava incelemek ve açık açık gemilerin ön ve yanlardan lenfatik damarları ve yağ incelemek için ince uçlu forseps kullanabilir ven için. </li><li> Aorta ve vena kava ve yavaş yavaş yayıldı forseps arasındaki doku uçağı bulun. Yavaş yavaş minimal travma ile doku yaymak için ince uçlu forseps ipuçları açın. </li><li> Açılı cezası geçerek renal arter üstün ve aşağı aorta etrafında gevşek 7 / Ç ipek kravatucu forseps gemilerin arkasına etrafında ve üzerinden bir dikiş çizim. Bu dikişler University of Wisconsin solüsyonu retrograd perfüzyon izin böbrek alma önce sıkılır. </li></ol> 2. Alıcı Hazırlık Donör hazırlığı takip başına 1,2-1,8 adımları. <ol><li> Aort, vena kava maruz operatörün sağ (Farenizin sol) için bağırsakları hareket ettirin. Steril tuzlu batırılmış bezle bağırsakları örtün. </li><li> Sağ renal arter bağlanarak bir sağ nefrektomi gerçekleştirin ve 7/0 ipek sütür ile birlikte ven ve sonra da bölmek. </li><li> 7 / Ç ipek sütür ve bölünmesine doğru üreter Arter. </li><li> Harekete geçirmek ve adım 1.14 ve 1.15 açıklandığı gibi tamamen renal arter ve ven aşağı aorta ve vena kava teşrih. Aorta ve vena kava arasında tam diseksiyonu olun. Vena kava ve aort anterior çalışan internal spermatik arteri korumak için özenBir lenfatik paket ile birlikte. </li><li> , Omurga vena kava ve damardan retroperitonda çalışan lomber damarlarını tanımlamak süreklilik 9 / Ç ipek ile lige, damarları bölmek için gerek yoktur. </li><li> Vasküler anastomozlar için arasındaki boşluk mikrovasküler kelepçeleri yerleştirmek için yeterli alan tanımlayın. </li><li> Dorsal penil ven yoluyla intravenöz heparin (5 adet) uygulayın. </li></ol> 3. Donör Böbrek Alma Arteriyel dolaşımın böbrek izole etmek alt ve üst aort etrafında yerleştirilmiş 7 / Ç ipek bağları sıkın. <ol><li> Aort içine bir iğne (31 g) ile University of Wisconsin çözeltisi, 0.5 ml - retrograd soğuk 0.2 infüze. </li><li> Dikişler içinde aorta bölmek ve böbrek ve aort uzunluğu ile renal arter kaldırmak için vena kava ile kavşakta renal ven bölün. Varsa w, aort kaynaklanan bel damarları Bölildirimde ligasyon. </li><li> Bir kültür kabı içinde steril bir pamuk ile soğuk tuzlu su içinde böbrek yerleştirin. </li><li> Servikal dislokasyon ile, verici bir fare öldürülür. </li></ol> 4. Böbrek Nakli - Böbrek Hazırlık <ol><li> Renal arter karşısında uzunlamasına doğrudan aort duvarı bölerek bir aort yama oluşturun. Arteriyel anastomoz yaparken bağlandı veya kaçınılması gereken yama herhangi bir damar lümen tanımlayın. </li><li> Inferiorda superiorda renal ven lümen ve ikinci ayrı dikiş dışarıdan içeriye bir 10 / Ç dikiş yerleştirin. Dikişler adrenal damar bölmek için kullanılır ve gonadal damar damar yönlendirmek için kullanılabilir. </li><li> Alıcı (Şekil 1.1) sağ kanadını böbrek yerleştirin ve dikişler de vasküler anastomozlar bezlerden veya diğer araçlar dolaşık olma dikişler olmadan tamamlanabilir sağlamak için yerleştirilir emin olun. </li></ol> 5. Böbrek Transplanlama - Vasküler Anastomoz İlk inferiorda sonra superiorda vena kava ve aorta kapsayan mikrovasküler kelepçeleri uygulayın. <ol><li> Vena kava ön duvarı delmek suretiyle bir iğne (31G) ile bir ventomy olun. % 0.9 NaCl yaklaşık 50 ul enjekte ederek vena kava yıkayın kan. </li><li> Uzunluğu, donor böbrek toplar damarı çapına denk olacak şekilde venotomi içinde açarak, ince uçlu forseps kullanılarak venotomi Widen. </li><li> Venotomi (Şekil 1.2) tepesinde birinci (renal ven lümeninde zaten) 10 / Ç üstün dikiş yerleştirin ve alt tepe ulaşana kadar (Şekil çalışan bir dikiş içinde anastomoz arka duvarını katılmak 1.3). </li><li> 10 / Ç alt dikiş yerleştirin ve sonra arka duvarından çalışan sütür kravat tek bir düğüm (Şekil 1.4) ilk kravat. </li><li> Ön wal oluşturmak alt dikiş kullanılarakanastomoz l çalışan bir dikiş ile (Şekil 1.5) ve üstün apeks (Şekil 1.6) de üstün dikiş sonuna kadar kravat. </li><li> Forseps ile aort duvarı çekme ve makas ile elips yama (yaklaşık aortotomy aortun çevresi ve uzunluğu üç kez renal arter lümeninin beşinci olmalı) keserek aortotomy oluşturun. </li><li> Içeriden dışarıdan aort yama üstün noktasında (Şekil 1.7) bir 10 / Ç dikiş yerleştirin ve aortotomy üstün tepesinde gemilere dışında kravat aorta geçer. </li><li> Çok sıkı sütür (Şekil 1.9) bağlayarak anastomoz daraltmak özen, çalışan bir dikiş superiorda alıcı aorta donör yama anastomoz (Şekil 1.8) başlayan arteriyel anastomoz oluşturun. </li><li> (Böbrek reperfüze sonra ilk üstün kelepçesini alt damar kelepçeleri sökünŞekil 1.10 ve 1.11). Üreter yeterli kan temini sağlandığı takdirde üreter Görünür peristaltizmin belki görülmektedir. </li></ol> 6. Böbrek Nakli - Üreterden Anastomoz <ol><li> Karın duvarından mesane ekleri bölün. </li><li> Her iki duvar aracılığıyla mesane sağa soldan iğne (21G) geçirin. </li><li> Iğne lümeninde düz forcep ipuçları yerleştirin ve geçmek hem de geri forseps artık mesanenin sol tarafında dışarı sağdan sola geçiyoruz şekilde mesane yoluyla. </li><li> Üreter sağ tarafında dışarı sonra sol mesane defekt geçecek şekilde mesane yoluyla donör üreter üzerinde dikiş çizin. </li><li> Yuvarlak gövdeli iğne 9 / Ç naylon sütür ile giriş noktası etrafında üç tek sütürler ile mesane adventisiyal üreter adventisya dikin. </li><li> Düğümlendikten için üreter proksimalinde kesti, böylece açılış inciE üreter idrar akışı ve üreter uç mesane vücuda geri çekilmesi için elde edilmesine olanak tanıyacak. Idrar Görünür üretim peri-üreter damarlardan kanama ile birlikte görülebilir. </li><li> Tek bir kesintiye 9 / Ç ipek sütür ile sağ yan mesane defekti kapatmak. </li></ol> 7. Kurtarma ve Post-operatif Bakım Orijinal yönde karın içine gastrointestinal organlara değiştirin ve 6 / Ç emilebilir sütür ile rektus kasları yaklaştırmayı karın duvarı kapatmak. <ol><li> Metal deri klipler ile cildi yaklaşır. </li><li> Kısmen atipamazole hidroklorür (10 g / ml) bir deri altından enjeksiyon ile anestezi ters. </li><li> Deri altından% 0.9 NaCl, 1 ml deri altından buprenorfin hidroklorür (0.05 mg / kg) ile ve akışkan destek enjeksiyon için analjezi uygulayın. </li><li> 24-48 saat boyunca 28 ° C&#39;de ısıtıldıktan kabininde fare kurtarmak. Için hastalık up fareyi gözlemleyinDeneyin tamamlanması. </li><li> 10 gün postoperativ - metal deri kliplerini 7 çıkartın. </li><li> Deney tamamlandıktan sonra, servikal dislokasyon ile fare euthanize. </li><li> Çıkarın ve histolojik analiz için, karşı böbrek allograftı ve toplar. </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>

Yazarlar ifşa hiçbir şey yok.

Arteriyel anastomoz gerçekleştirmek için en iyi tarif şekilde alıcı aorta uç-yan bir şekilde, devamında renal arter ile, donörün uzak aorta kullanmaktır. Daha uygun olması için inanıyoruz insan böbrek nakli yapılan &#39;Carrell ve yama &quot;ikizleme benzer bir aort yama, kullanımını tarif etmektedir. Donör ve alıcı operasyon süresi literatürde raporlar seyrek olsa biz alıcı aorta için uç-yan donör aort yerine alıcı aort, aort yama kullanan tercih olduğuna inanıyorum. Bir yama kullanara...

Uzunluk ve yaşam 2 kalitesinde iki gelişme sunan, dünya çapında son dönem böbrek yetmezliği olan hastalar için devrimci bir tedavi haline gelmiştir o zamandan bu yana böbrek yetmezliği tedavisi için başarılı böbrek nakli ilk, tek yumurta ikizleri 1 arasında 1955 yılında tarif edilmiştir. Ancak uzun dönem greft sağkalım kronik allogreft hasar 3 sonuçlanan patolojik süreçlerin bir çok engel olmuştur. İnsanlarda nakledilen böbreğin reddedilmesi immunosupporessive rejimlerinde önemli gelişmelere rağmen, önemli bir morbidite nedenidir. Böbrek nakli, bir fare modeli geliştirilmesi amacı yakından insan renal transplantasyon 4 bulundu süreci ve patolojiyi çoğaltmak. Skoskiewicz ve ark., 1973 5 böbrek nakli fare modelini tarif. Gelişmiş mikrocerrahi becerileri gerekli olmasına rağmen, değerli bir tçeşitli nedenlerden dolayı ool: fare genomu iyi karakterize ve deneysel yöntemler ve fare çalışmaları için mevcut tekniklerin büyük bir çeşitlilik söz konusudur olmuştur. Böbrek nakli fare modeli kullanılarak çok grup, ancak deney 4 süresince yerinde bırakılır diğer çalışmalarda ve alıcı farelerin saf böbreklerin bizim tarif edilen metodoloji, bir de, hayatı destekleyen organ olarak böbrek nakli kullandık. Yararı, fare ve böylece fare morbidite ve ikinci bir prosedür ölüm riskini azaltan, tek ve anestezi sergilemesidir. Ayrıca fare kademeli böbrek yetmezliği olumsuz etkilerinden muzdarip değil. Allojenik ret modelleri, kalp ve deri gibi diğer organlarda mevcut olmasına rağmen, bu her zaman böbrek nakli ile doğrudan ilgili değildir. Bu modeller farklı modları ve dy ortaya kanıtlar vardırreddinin iç- sel, örneğin kalp alograft ve böbrek nakli reddi, zaman süreci, belirli gerilme kombinasyonları 6 önemli ölçüde farklıdır. Bu transjenik olmayan FVB / NJ farelerine, BALB / c donörlerinde akut renal alograft reddi, kalıpları tarif etmişlerdir, bu model T hücreleri ve makrofajlar 7 birikimi ile hücresel kaynaklı yaralanma sergiledi. Bu farenin tek bir MHC sınıf II loci mis ile karakterizedir Alternatif olarak biz de interstisyel fibrozis ve tübüler atrofi sergileyen kronik allogreft hasar modeli tarif var, bu, C57BL / 6 alıcılar içine C57BL / 6 BM12 bağışçıları bir böbrek nakli sonucu -match 8. Naklinin çok yönlü olarak akut rejeksiyon, hücresel ve hümoral rejeksiyon, iskemi reperfüzyon yaralanması, ve yeni tedavi edici ajanlar da dahil olmak üzere ana hatlarını açıklar böbrek nakli fare modeli kullanarak çalışılmıştır. Biz cerrahi t değiştirdinizechnique çalışma süresini azaltmak ve cerrahi kolaylığını artırmak için. Özellikle simultane donör ve alıcı hazırlık ve sürekli aort yama anastomoz kullanarak basitleştirilmiş bir vasküler anastomoz tekniğini tarif var. Bu video ve el yazması bu tekniğin kurulması yardımcı olmak için kilit noktaları sağlayacaktır.

<table border="0" cellpadding="0" cellspacing="0">
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			<td>Blunt Dissecting Scissors</td>
			<td>Fine Science Tools</td>
			<td>&#160;14072-10</td>
			<td>For skin cutting</td>
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			<td>Curved Castoviejo scissors</td>
			<td>Fine Science Tools</td>
			<td>15017-10</td>
			<td>For tissue cutting</td>
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			<td>Spring Scissors &#8211; straight</td>
			<td>Fine Science Tools</td>
			<td>15000-08</td>
			<td>For suture cutting</td>
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			<td>Toothed forceps 1x2 teeth</td>
			<td>Fine Science Tools</td>
			<td>11021-12</td>
			<td></td>
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			<td>2 x Fine Tip forceps (Dumont No.5)</td>
			<td>Fine Science Tools</td>
			<td>11251-20</td>
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			<td>Angled Fine Tip forceps (Dumont No. 5/45)</td>
			<td>Fine Science Tools</td>
			<td>11253-25</td>
			<td>For blunt dissecting</td>
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			<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>
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			<td>Fine Science Tools</td>
			<td>1300-04</td>
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			<td>Fine Science Tools</td>
			<td>18055-04</td>
			<td>Microvascular clamps</td>
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			<td>2 x Colibri 3cm wire retractor</td>
			<td>Fine Science Tools</td>
			<td>17000-03</td>
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			<td>Castroviejo needle holder with lock</td>
			<td>Fine Science Tools</td>
			<td>120660-01</td>
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			<td>12031-07</td>
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			<td>7mm wound clips</td>
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			<td>12032-07</td>
			<td>Remove 7 to 10 days after surgery</td>
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			<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.

Renal alograft reddi, nakledilen böbreğin (Şekil 2) metakarn sabitlenmiş parafine gömülü doku bölümleri histolojik analiz ile değerlendirilebilir. Renal iskemi reperfüzyon hasarı sinjeneiktir farenin sonuçları arasında böbreklerin Isograft nakli, ancak 4 hafta tübüller kurtarıldı ve yerli böbrekler için histolojik karşılaştırılabilir var. Akut rejeksiyon böbrek parankimi genelinde yaygın mononükleer hücre infiltrasyonu İnterstisyum, glomerüller ve tübüllerini içeren, ...

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.

Fare Böbrek Transplantasyonu: Allogreft Reddi Modelleri

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 Görüntüleme.  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.

Video: 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.

Murin Islet İzolasyon ve subkapsüler Böbrek Nakli Bir Yöntem

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

Tıp

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.

Sıçan Böbrek Nakli sonra Akut Allogreft Reddi non-invaziv Görüntüleme Kullanımı<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 ...

High-frequency Ultrasound Imaging of Mouse Cervical Lymph Nodes

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal systems. HFUS has the ability to detect structures as small as 30 &#181;m, a property that has been utilized for visualizing superficial lymph nodes in rodents in brightness (B)-mode. Combining power Doppler with B-mode imaging allows for measuring circulatory blood flow within lymph nodes and other organs. While HFUS has been utilized for lymph node imaging in a number of mouse&#160; model systems, a detailed protocol describing HFUS imaging and characterization of the cervical lymph nodes in mice has not been reported. Here, we show that HFUS can be adapted to detect and characterize cervical lymph nodes in mice. Combined B-mode and power Doppler imaging can be used to detect increases in blood flow in immunologically-enlarged cervical nodes. We also describe the use of B-mode imaging to conduct fine needle biopsies of cervical lymph nodes to retrieve lymph tissue for histological&#160; analysis. Finally, software-aided steps are described to calculate changes in lymph node volume and to visualize changes in lymph node morphology following image reconstruction. The ability to visually monitor changes in cervical lymph node biology over time provides a simple and powerful technique for the non-invasive monitoring of cervical lymph node alterations in preclinical mouse models of oral cavity disease.

This protocol describes the application of high-frequency ultrasound (HFUS) for imaging mouse cervical lymph nodes. This technique optimizes visualization and quantification of cervical lymph node morphology, volume and blood flow. Image-guided biopsy of cervical lymph nodes and processing of lymph tissue for histological evaluation is also demonstrated.

Fare Servikal lenf nodu Yüksek frekanslı Ultrasonik Görüntüleme

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal ...

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.

Aloimmün kaynaklı Vasküler Reddi ve Nakli arterioskleroz Fare Modeli

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.

Murin Böbrek nakli Tekniği

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.

Normotermik<em&gt; Ex Vivo</em&gt; Böbrek Koruma Böbrek Nakli Perfüzyon önce greftler

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.

Fare modelleri Glomerular hastalığı böbrek fonksiyonlarının değerlendirilmesi

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.

Ortootopik Rat böbrek nakli: bir roman ve Basitleştirilmiş cerrahi yaklaşım

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.

Optik temizleme ve Immünolabeled böbrek dokusu görüntüleme

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.