Nrl-GFP fareleri sağladığı için Anand Swaroop'a, teknik destek için Jochen Haas'a ve hayvancılık için Sindy Böhme ve Emely Lessmann'a teşekkür ederiz.	
	Bu çalışma Deutsche Forschungsgemeinschaft (DFG) tarafından desteklenmiştir: FZT 111 - Rejeneratif Terapiler Merkezi Dresden, CRTD Tohum Hibe Programı, SFB 655 ve ProRetina e.V. vakıf, DIGS-BB Lisansüstü Programı Dresden ve Fundação para a Ciência e Tecnologia (SFRH/BD/60787/2009)

Hayvanların etik kullanımı ve bakımı beyanı:
Tüm hayvan deneyleri Avrupa Birliği ve Alman yasalarına (Tierschutzgesetz) sıkı bir şekilde uygun olarak gerçekleştirildi ve ARVO Oftalmik ve Vizyon Araştırmalarında Hayvanların Kullanımı Beyanı'na bağlı kaldı. Tüm hayvan deneyleri TU Dresden ve Landesdirektion Dresden hayvan etik komitesi tarafından onaylanmıştır (onay numarası: 24D-9168.11-1/2008-33).
1. Hücre Ayrıştırma ve Hücre Sıralamaya Başlamadan Önce
<ol><li>Üç 15 ml reaksiyon tüpünü şu şekilde etiketle: Yıkama (W), Pozitif fraksiyon (+) ve Negatif fraksiyon (-).</li>
</ol>2. Retina Ayrışması
<ol><li>Bir makas kullanarak PN 4 yavrularının kafasını koparın.</li>
	<li>Yavruların başını kısa bir süre içinde% 70 etanolde durulayın ve ardından PBS'de yıkayın.</li>
	<li>Başı soğuk HBSS'ye aktarın ve gözü enükle edin (bu adımı tüm kafalar için tekrarlayın). Enükle etmek için göz kapaklarını açın ve gözü optik sinir bölgesinde kavisli tokmaklarla sabitlayın. Sonra gözü dikkatlice yörüngeden çıkarın.</li>
	<li>Tüm gözleri kaplattıktan sonra retinayı izole edin: optik sinire kapalı bir makas sokun ve bıçakları açın. Peal RPE/chorid dışarı. Kavisli kümesleri kullanarak lensleri ve kan damarlarını çıkarın.</li>
	<li>İzole retinaları papain çözeltisini içeren 1,5 ml reaksiyon tüpüne aktarın (papain ayrışma kiti tarafından sağlanır).</li>
	<li>Retinaları papain çözeltisinde 37 °C'de bir su banyosunda veya çalkalayıcıda (400 rpm) 30-60 dakika kuluçkaya yatırın.<ol>
<li>Retinalar papain çözeltisinde kuluçkaya yatarken, pipet 15 ml reaksiyon tüpüne (etiket "1") 1 ml ovomucoid çözeltisi.</li>
			<li>15 ml reaksiyon tüpüne (etiket "2") 60 μl DNase I (10 mg/ml) + 60 μl ovomucoid çözeltisi (kit tarafından sağlanır) + 520 μl EBSS ekleyin.</li>
		</ol>
</li>
	<li>Papain inkübasyonundan sonra, kısmen sindirilmiş retinaları içeren papain çözeltisini reaksiyon tüpü "2"ye aktarın.</li>
	<li>Yangın cilalı pipet kullanarak mekanik ayrışma (10x yukarı ve aşağı) gerçekleştirin.</li>
	<li>Pipet tek hücre süspansiyon reaksiyon tüpü "1". Hücre süspansiyonu ovomucoid çözeltisinin üzerine hafifçe katlayarak 2 katman oluşturmaya çalışın.</li>
	<li>300 x g'da (yaklaşık 1.300 rpm) 5 dakika santrifüj.</li>
	<li>Üstnatant atın ve hücreleri 500 μl MACS tamponunda yeniden biriktirin.</li>
</ol>3. Manyetik İlişkili Hücre Sıralama (MACS) Kullanarak Hücre Sıralama
<ol><li>10 μg/ ml'lik son konsantrasyonu elde etmek için 500 μl'ye X μl sıçan anti-CD73 antikoru ekleyin.</li>
	<li>Buzda 5 dakika kuluçkaya yaslanın.</li>
	<li>15 ml reaksiyon tüpünü MACS tamponu ile 10 ml'ye doldurun ve 300 x g hızında 5 dakika santrifüj edin.</li>
	<li>Üsttekini çıkarın.</li>
	<li>Peleti 480 μl MACS tamponunda yeniden sakla ve 120 μl keçi anti-sıçan IgG mikrobead ekleyin.</li>
	<li>Buz üzerinde 15 dakika kuluçkaya yatır; karıştırmayın, sallamayın veya karıştırmayın.</li>
	<li>15 ml reaksiyon tüpünü MACS tamponu ile 5 ml'ye doldurun ve 300 x g'da 5 dakika santrifüj edin.</li>
	<li>Reaksiyon tüpü santrifüjlendikçe:<ol>
<li>LS Sütununu manyetik standa ayarlayın.</li>
			<li>Önsezi filtresini LS Sütunu'nun üstüne yerleştirin.</li>
			<li>Önsezim filtresini ve LS Sütununu 3 ml MACS tamponu ile nemlendirin ve MACS tamponunu yıkama (W) tüpüne toplayın.</li>
		</ol>
</li>
	<li>Üsttekini çıkarın.</li>
	<li>Peletin 500 μl MACS tamponunda yeniden süzülür.</li>
	<li>500 μl hücre süspansiyonunu filtreye yükleyin, ardından filtreye 1 ml MACS tamponu ekleyin ve negatif fraksiyonu negatif fraksiyon (-) reaksiyon tüpüne toplayın.</li>
	<li>Ardından, sütuna bağlı olmayan hücreleri yıkamak için sütuna 3 x 3 ml MACS arabelleği ekleyin</li>
	<li>Bu 9 ml LS Sütunundan geçtikten sonra, sütunu manyetik standdan çıkarın ve pozitif fraksiyon (+) reaksiyon tüpünün üzerine takın.</li>
	<li>LS Sütununa hızlı bir şekilde 5 ml MACS arabelleği yükleyin ve pistonu LS Sütununa koyun ve tüm arabellek sütundan geçene kadar bastırın.</li>
	<li>Pozitif fraksiyonu içeren reaksiyon tüpünü 300 x g'da 5 dakika santrifüj edin.</li>
	<li>Üst düğmeyi çıkarın, 500 μl MACS tamponunda yeniden dirilin ve 4 °C'de tutun.</li>
	<li>Ortak bir hücre sayma aygıtı kullanarak toplam hücre miktarını sayın.</li>
	<li>MACS tamponunda 2 x 105 hücre/μl içeren pozitif sıralanmış fraksiyondan bir hücre süspansiyonu hazırlayın ve 4 °C'de tutun</li>
</ol>4. MAC sıralı Fotoreceptör Öncü Hücrelerin Fare Retinasına Nakli
<ol><li>Aşağıdaki çözümlerin tümünde hazır olduğundan emin olun: 1 ml steril PBS veya HBSS, 10 μl DNASE I, 1-2 ml steril deiyonize H2O, hücre süspansiyon fraksiyonlarının aliquots 4 °C.</li>
	<li>Yetişkin fareyi(yani 2-4 aylık) medetomidin hidroklorür intraperitoneal enjeksiyonu (0.01 mg/10 g vücut ağırlığı), ketamin (0.75 mg/10 g vücut ağırlığı) ile uyuşturun. Daha sonra ağrı kesici için buprenorfin (0.05 mg / kg vücut ağırlığı) deri altı enjeksiyonu yapın.</li>
	<li>Fenilofilin düşüşü olan genişlemiş gözbebekleri %2.5-Tropicamid %0.5.</li>
	<li>Fareyi fare başı tutucusuna sabitle ve stereo mikroskop altına yerleştirin.</li>
	<li>Gözün kurumasını önlemek için bir damla Vizidik jel uygulayın.</li>
	<li>İğnede steril 30 G 1/2 kullanarak sklera ve kornea (sırasıyla ora serrata)arasındaki sınırda küçük bir delik açın.</li>
	<li>Ortak bir kapak kaydıramasını elmas bir kalem kullanarak küçük (yaklaşık 5 mm x 5 mm) parçalar halinde kesin, bu parçalardan birini korneanın üzerine yerleştirin ve retinanın doğrudan görselleştirilmesini sağlar.</li>
	<li>Presterilize mikroliter şırıngasını deiyonize su ile birkaç kez yıkayın.</li>
	<li>Mikroliter şırıngayı 1 μl hücre süspansiyonu ile yükleyin, konjonktiva ve sklera yoluyla teğetsel olarak doğrudan iğne yapın ve retinanın burun yarısında görsel kontrol altına yerleştirin.</li>
	<li>Alt alana ulaşana kadar retinaya hafifçe bir delik açın, hücre süspansiyonu alt alana enjekte edin.</li>
	<li>Retina alanının yaklaşık 1/4'lerini kanama olmadan kaplayan, düzgün olması gereken retinanın büllöz kopuşunun gözlemlenmesi.</li>
	<li>Şırındıcı nazikçe çekin, retina deliği otomatik olarak kapanır.</li>
	<li>Mikroliter şırıngasını deiyonize suyla birkaç kez yıkayın.</li>
	<li>Fareyi baş tutucudan serbest bırakın.</li>
	<li>Medetomidin hidroklorür etkisinin tersine çevrilmesi için atipamozol hidroklorür enjekte ederek fareyi uyandırın.</li>
	<li>Uyanma süresi için fareyi sıcak karanlık odaya (yaklaşık 25 °C) yerleştirin.</li>
	<li>Transplantasyondan sonraki 2 gün sonra, Ağrı kesici için sabah ve öğleden sonra deri altı enjeksiyonu ile Buprenorfin (0.05 mg/kg vücut ağırlığı) uygulanmalıdır.</li>
</ol>

<ol>
	<li>Bartsch, U., 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=Retinal+cells+integrate+into+the+outer+nuclear+layer+and+differentiate+into+mature+photoreceptors+after+subretinal+transplantation+into+adult+mice.">Retinal cells integrate into the outer nuclear layer and differentiate into mature photoreceptors after subretinal transplantation into adult mice.</a> Exp. Eye Res. 86, 691-700 (2008).</li><li>MacLaren, R. E., 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=Retinal+repair+by+transplantation+of+photoreceptor+precursors.">Retinal repair by transplantation of photoreceptor precursors.</a> Nature. 444, 203-207 (2006).</li><li>Eberle, D., 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=Outer+segment+formation+of+transplanted+photoreceptor+precursor+cells.">Outer segment formation of transplanted photoreceptor precursor cells.</a> PLoS One. 7, (2012).</li><li>Lakowski, 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=Cone+and+rod+photoreceptor+transplantation+in+models+of+the+childhood+retinopathy+Leber+congenital+amaurosis+using+flow-sorted+Crx-positive+donor+cells.">Cone and rod photoreceptor transplantation in models of the childhood retinopathy Leber congenital amaurosis using flow-sorted Crx-positive donor cells.</a> Hum. Mol. Genet. 19, 4545-4559 (2010).</li><li>Barber, A. 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=Repair+of+the+degenerate+retina+by+photoreceptor+transplantation.">Repair of the degenerate retina by photoreceptor transplantation.</a> Proc. Natl. Acad. Sci. U.S.A. 110, 354-359 (2013).</li><li>Pearson, 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=Restoration+of+vision+after+transplantation+of+photoreceptors.">Restoration of vision after transplantation of photoreceptors.</a> Nature. , (2012).</li><li>Singh, M. 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=Reversal+of+end-stage+retinal+degeneration+and+restoration+of+visual+function+by+photoreceptor+transplantation.">Reversal of end-stage retinal degeneration and restoration of visual function by photoreceptor transplantation.</a> Proc. Natl. Acad. Sci. U.S.A. 110, (2013).</li><li>Eberle, D., Schubert, S., Postel, K., Corbeil, D., Ader, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+integration+of+transplanted+CD73-positive+photoreceptor+precursors+into+adult+mouse+retina.">Increased integration of transplanted CD73-positive photoreceptor precursors into adult mouse retina.</a> Invest. Ophthalmol. Vis. Sci. 52, 6462-6471 (2011).</li><li>Koso, 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=CD73,+a+novel+cell+surface+antigen+that+characterizes+retinal+photoreceptor+precursor+cells.">CD73, a novel cell surface antigen that characterizes retinal photoreceptor precursor cells.</a> Invest. Ophthalmol. Vis. Sci. 50, 5411-5418 (2009).</li><li>Lakowski, 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=Effective+transplantation+of+photoreceptor+precursor+cells+selected+via+cell+surface+antigen+expression.">Effective transplantation of photoreceptor precursor cells selected via cell surface antigen expression.</a> Stem Cells. 29, 1391-1404 (2011).</li><li>Akimoto, 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=Targeting+of+GFP+to+newborn+rods+by+Nrl+promoter+and+temporal+expression+profiling+of+flow-sorted+photoreceptors.">Targeting of GFP to newborn rods by Nrl promoter and temporal expression profiling of flow-sorted photoreceptors.</a> Proc. Natl. Acad. Sci. U.S.A. 103, 3890-3895 (2006).</li><li>Eiraku, 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=Self-organizing+optic-cup+morphogenesis+in+three-dimensional+culture.">Self-organizing optic-cup morphogenesis in three-dimensional culture.</a> Nature. 472, 51-56 (2011).</li><li>Nakano, 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=Self-formation+of+optic+cups+and+storable+stratified+neural+retina+from+human+ESCs.">Self-formation of optic cups and storable stratified neural retina from human ESCs.</a> Cell Stem Cell. 10, 771-785 (2012).</li><li>Osakada, 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=Toward+the+generation+of+rod+and+cone+photoreceptors+from+mouse,+monkey+and+human+embryonic+stem+cells.">Toward the generation of rod and cone photoreceptors from mouse, monkey and human embryonic stem cells.</a> Nat. Biotechnol. 26, 215-224 (2008).</li><li>Lee, M. Y., Lufkin, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+the+&amp;#34;Three-step+MACS&amp;#34;:+a+novel+strategy+for+isolating+rare+cell+populations+in+the+absence+of+known+cell+surface+markers+from+complex+animal+tissue.">Development of the &amp;#34;Three-step MACS&amp;#34;: a novel strategy for isolating rare cell populations in the absence of known cell surface markers from complex animal tissue.</a> J. Biomol. Tech. 23, 69-77 (2012).</li></ol>

Yazarlar rakip finansal çıkarlar beyan etmemektedir.

Fotoreceptör öncül hücrelerin subretinal nakli, bu ışığa duyarlı hücrelerin ana bilgisayar retinalarına önemli sayılarda entegrasyonunu sağlamak için güvenilir bir aracı temsil eder1,2. Bu, gelecekte retina dejeneratif hastalıkların tedavisi için bir hücre tedavisi kurulmasına izin verebilir6. Şu anda PN 4 retinalarından izole edilmiş olan hücrelerin donör popülasyonu, subretinal enjeksiyondan sonra sadece fotoreceptör öncül hücrelerin entegre olduğu farklı hücre türlerinin bir karı?...

Görme, insanların en önemli duyularından biridir. Bu duygunun bozulması ve körlük, sanayileşmiş ülkelerde engelliliğin başlıca nedenlerinden biridir. Görme bozukluğu veya körlüğün baskın nedeni, makula dejenerasyonu, retinitis pigmentosa, koni-çubuk distrofisi ve diğer durumlarda gözlenebildiği gibi fotoreceptör hücre kaybı ile karakterize retina dejenerasyonudur. Bugüne kadar, kayıp görüşü geri yüklemek için etkili bir tedavi mevcut değildir. 2006 ve 2008'de birbirinden bağımsız iki farklı laboratuvar rapor edildi, çubuk fotoreceptör öncül hücrelerinin yetişkin vahşi tip fare retinalarına başarılı bir şekilde nakli1,2. Böylece, fotoreceptör öncüsü hücre nakli olasılığının da dejenere bir retinaya dönüşmesi, dejenere fotoreceptörlerin yerini alması ve görmeyi geri kazanma olasılığı ortaya çıkmıştır. Nitekim, son zamanlarda gösterilmiştir. bu tür nakledilen fotoreceptör öncül hücrelerin, düzgün bir şekilde geliştirilen dış segmentler3, endojen bipolar hücrelere yakın sinaptik terminaller ve dış nükleer katman2-4'tebulunan yuvarlak bir hücre gövdesi gibi olgun vahşi tip fotoreceptörlerin morfolojik kriterlerini ortaya çıkardığı ve ayrıca5-7ana sinir devresine işlevsel olarak entegre olma yeteneği . Bu stratejinin ana ilkelerinden biri, doğum sonrası 4. gün (PN 4, PN0 doğum günü olarak tanımlanır) genç fare retinalarının kullanılmasıdır ve bu da transplantasyon için farklı hücre tiplerinin bir karışımı ile sonuçlanır. Gelecekteki bir terapötik uygulamanın arka planında, bu karışımın fotoreceptör öncül hücreler için saflaştırılması gerekir. CD73 retina8-10genç fotoreceptörler için özel ilk hücre yüzey belirteci olarak tanımlanmıştır. Burada, bu hücre yüzeyi işaretleyicisine dayanan ve manyetik ilişkili hücre sıralama (MACS) tekniğinin kullanıldığı bir fotoreceptör öncül hücre temizleme yöntemini gösteriyoruz. MACS, hızlı sıralama süreleri ve GMP koşullarına daha kolay uyum sağladığı için floresanla etkinleştirilen hücre sıralama tekniklerine kıyasla avantajlara sahip olabilir. Zenginleştirilmiş popülasyonu yetişkin vahşi tip retinalarda alt alana nakledirken ~%90 zenginleştirme ve 3 kata kadar daha yüksek entegrasyon oranı gösterebiliriz. Bu nedenle, MACS tabanlı fotoreceptör öncül hücre zenginleştirme ve subretinal transplantasyon, retina dejenerasyonu tedavisi için rejeneratif terapötik stratejinin geliştirilmesi için güvenilir ve umut verici tekniklerdir.

<table border="0" cellpadding="0" cellspacing="0" width="585">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="60">
			<td height="60" style="height:60px;width:195px;">Papain Dissociation System</td>
			<td style="width:111px;">Worthington&#160; Biochemical Corporation</td>
			<td style="width:103px;">LK003150</td>
			<td style="width:176px;">supplied DNase I is not used in the method</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">purified rat anti-mouse CD73, clone TY/23</td>
			<td style="width:111px;">BD Pharmingen</td>
			<td style="width:103px;">550738</td>
			<td style="width:176px;">Stock concentration 0.5mg/ml</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Goat Anti-Rat IgG MicroBeads</td>
			<td style="width:111px;">Miltenyi</td>
			<td style="width:103px;">130-048-501</td>
			<td style="width:176px;">Total volume of 2ml</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">PBS</td>
			<td style="width:111px;">Gibco</td>
			<td style="width:103px;">10010-015</td>
			<td style="width:176px;">Used to count the total number of cells</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">DNase I</td>
			<td style="width:111px;">Sigma</td>
			<td style="width:103px;">D5025-150KU</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">HBSS</td>
			<td style="width:111px;">Gibco</td>
			<td style="width:103px;">14025050</td>
			<td style="width:176px;">Used for dissociation of the retinas</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Trypan blue</td>
			<td style="width:111px;">Sigma</td>
			<td style="width:103px;">Fluka93595</td>
			<td style="width:176px;">Used to count the total number of cells</td>
		</tr>
		<tr height="100">
			<td height="100" style="height:100px;width:195px;">Vidisic</td>
			<td style="width:111px;">Dr. Mann Pharma / Andreae-Noris Zahn AG</td>
			<td style="width:103px;">-</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">Domitor</td>
			<td style="width:111px;">Pfizer</td>
			<td style="width:103px;">76579</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">Ketamin 10%</td>
			<td style="width:111px;">Ratiopharm</td>
			<td style="width:103px;">7538843</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">Antisedan</td>
			<td style="width:111px;">Pfizer</td>
			<td style="width:103px;">76590</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="81">
			<td height="81" style="height:81px;width:195px;">Phenylephrin 2.5%-Tropicamid 0.5%</td>
			<td style="width:111px;">University clinics Dresden pharmacy</td>
			<td style="width:103px;">-</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:195px;"></td>
			<td style="width:111px;"></td>
			<td style="width:103px;"></td>
			<td style="width:176px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:195px;">Name of Equipment</td>
			<td style="width:111px;">Company</td>
			<td style="width:103px;">Catalog Number</td>
			<td style="width:176px;">Comments/Description</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">Pre-Separation Filters</td>
			<td style="width:111px;">Miltenyi</td>
			<td style="width:103px;">130-041-407</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">LS Columns</td>
			<td style="width:111px;">Miltenyi</td>
			<td style="width:103px;">130-042-401</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">MACS MultiStand</td>
			<td style="width:111px;">Miltenyi</td>
			<td style="width:103px;">130-042-303</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">QuadroMACS Separator</td>
			<td style="width:111px;">Miltenyi</td>
			<td style="width:103px;">130-090-976</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:195px;">fire polish glass pasteur pipette</td>
			<td style="width:111px;">Brand</td>
			<td style="width:103px;">74777 20</td>
			<td style="width:176px;">The pipette&#8217;s tips need to be fire-polished and autoclaved.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">MACS 15ml tube rack</td>
			<td style="width:111px;">Miltenyi</td>
			<td style="width:103px;">130-091-052</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">Cell count chamber</td>
			<td style="width:111px;">Carl Roth</td>
			<td style="width:103px;">T728.1</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Sterile 15ml tubes</td>
			<td style="width:111px;">Greiner Bio-one</td>
			<td style="width:103px;">188271</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Leica M651 MSD</td>
			<td style="width:111px;">Leica</td>
			<td style="width:103px;">M651 MSD</td>
			<td style="width:176px;">can be used instead of Olympus SZX10</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Olympus SZX10</td>
			<td style="width:111px;">Olympus</td>
			<td style="width:103px;">SZX10</td>
			<td style="width:176px;">can be used instead of Leica M651 MSD</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Olympus inverted stereo microscope CKX41</td>
			<td style="width:111px;">Olympus</td>
			<td style="width:103px;">CKX41</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Cell culture hood Thermo Scientific MSC-Advance</td>
			<td style="width:111px;">Thermo scientific</td>
			<td style="width:103px;">51025411</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">1.5ml reaction tube</td>
			<td style="width:111px;">Sarstedt</td>
			<td style="width:103px;">727706400</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">2ml reaction tube</td>
			<td style="width:111px;">Sarstedt</td>
			<td style="width:103px;">72695</td>
			<td style="width:176px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Eppendorf Centrifuge 5702</td>
			<td style="width:111px;">VWR (Eppendorf)</td>
			<td style="width:103px;">521-0733</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">Mouse head holder</td>
			<td style="width:111px;">myNeurolab</td>
			<td style="width:103px;">471030</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">BD Microlance 3 30G 1/2&#8221;</td>
			<td style="width:111px;">BD Pharmingen</td>
			<td style="width:103px;">304000</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Hamilton microliter syringe 5&#181;l, 75RN</td>
			<td style="width:111px;">Hamilton</td>
			<td style="width:103px;">065-7634-01</td>
			<td style="width:176px;">delivered without needles</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Hamilton RN special needle GA34</td>
			<td style="width:111px;">Hamilton</td>
			<td style="width:103px;">065-207434</td>
			<td style="width:176px;">Blunt, 12mm length</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:195px;">Vannas-T&#252;bingen Spring Scissors - 5mm Blades Straight</td>
			<td style="width:111px;">Fine Science Tools</td>
			<td style="width:103px;">15003-08</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:195px;">Dumont #7 Forceps - Titanium Biologie</td>
			<td style="width:111px;">Fine Science Tools</td>
			<td style="width:103px;">11272-40</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:195px;">Diamond pen</td>
			<td style="width:111px;">Tools-tech</td>
			<td style="width:103px;">-</td>
			<td style="width:176px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:195px;">15x15mm Cover slips</td>
			<td style="width:111px;">Sparks</td>
			<td style="width:103px;">MIC3366</td>
			<td style="width:176px;">-</td>
		</tr>
	</tbody>
</table>

subretinal transplantation of macs purified photoreceptor precursor cells into the adult mouse retina

Retinanın ışık algılayan hücrelerinin, yani fotoreceptörlerin kaybına bağlı görme bozukluğu ve körlük, sanayileşmiş ülkelerde temel engellilik nedenini temsil eder. Dejenere fotoreceptörlerin hücre nakli ile değiştirilmesi, gelecekteki klinik uygulamalarda olası bir tedavi seçeneğini temsil etmektedir. Nitekim, son preklinik çalışmalar, doğum sonrası 4. günde yenidoğan fare retinasından izole edilen olgunlaşmamış fotoreceptörlerin subretinal transplantasyondan sonra yetişkin fare retinasına entegre olma potansiyeline sahip olduğunu göstermiştir. Donör hücreler, iç ve dış segmentler, dış nükleer tabakada bulunan yuvarlak bir hücre gövdesi ve endojen bipolar hücrelere yakın sinaptik terminaller dahil olmak üzere olgun bir fotoreceptör morfolojisi oluşturdu. Nitekim, son raporlar donör fotoreceptörlerin konak farelerin sinir devrelerine işlevsel olarak entegre olduğunu göstermiştir. Bu tür hücre değiştirme yaklaşımının gelecekteki bir klinik uygulaması için, tercih edilen hücrelerin saflaştırılmış süspansiyonlarının üretilmesi ve göze uygun entegrasyon için doğru konuma yerleştirilmesi gerekir. Fotoreceptör öncüllerinin zenginleştirilmesi için, donör hücrelerin genetik muhabir modifikasyonunun önlenmesi için sıralama belirli hücre yüzeyi antijenlerine dayanmalıdır. Burada manyetik ilişkili hücre sıralamasını (MACS) gösteriyoruz - hücre yüzeyi işaretleyici CD73'e dayanarak fotoreceptöre özgü muhabir farelerin yenidoğan retinasından izole edilen nakledilebilir çubuk fotoreceptör öncüllerinin zenginleştirilmesi. Anti-CD73 antikorları ile inkübasyon ve ardından mikro boncuk konjuge sekonder antikorlar, çubuk fotoreceptör öncüllerinin MACS tarafından zenginleştirilmesini yaklaşık% 90'a kadar sağladı. Akış sitometrisine kıyasla MACS, GMP standartlarına daha kolay uygulanabilmesi ve yüksek miktarda hücrenin nispeten kısa zaman periyotlarında sıralanabilmesi avantajına sahiptir. Zenginleştirilmiş hücre süspansiyonlarının yetişkin vahşi tip farelerin alt alanına enjekte etmesi, sıralanmamış hücre süspansiyonlarına kıyasla 3 kat daha yüksek entegrasyon oranına neden oldu.

Çubuk fotoreseptörlerin fare retinasına entegre olma yeteneğini değerlendirmek için, GFP'nin nöral retina lösin fermuarı (Nrl, Nrl-GFP) promotörü11tarafından yönlendirildiği bir fare muhabir hattı kullanılmıştır. Nrl, yetişkinlik boyunca E12.5'te ekspresyonunu başlatan ve donör çubuk fotoreceptör hücrelerinin belirli bir şekilde etiketlenebilmesini sağlayan çubuk fotoreceptörlerin en erken belirtecidir.
PN 4 Nrl-GFP yavrularının kafası kesildi ve göz...

Watch this Scientific Journal Video about Subretinal Transplantation of MACS Purified Photoreceptor Precursor Cells into the Adult Mouse Retina at JoVE.com

Subretinal Transplantation of MACS Purified Photoreceptor Precursor Cells into the Adult Mouse Retina

Hücre nakli fotoreceptör kaybı ile karakterize retina dejenerasyonu tedavisi için bir stratejiyi temsil eder. Burada, nakledilebilir fotoreceptörlerin zenginleştirilmesi ve yetişkin farelere alt deri nakli için bir yöntem açıklıyoruz.

MACS Saflaştırılmış Fotoreceptör Öncü Hücrelerinin Yetişkin Fare Retinasına Subretinal Transplantasyonu

Vision impairment and blindness due to the loss of the light-sensing cells of the retina, i.e. photoreceptors, represents the main reason for disability ...

subretinal-transplantation-macs-purified-photoreceptor-precursor

Research

JoVE Journal

Medicine

10.6K Görüntüleme.  Technische Universität Dresden. Hücre nakli fotoreceptör kaybı ile karakterize retina dejenerasyonu tedavisi için bir stratejiyi temsil eder. Burada, nakledilebilir fotoreceptörlerin zenginleştirilmesi ve yetişkin farelere alt deri nakli için bir yöntem açıklıyoruz.

Video: Subretinal Transplantation of MACS Purified Photoreceptor Precursor Cells into the Adult Mouse Retina

Subretinal Injection of Gene Therapy Vectors and Stem Cells in the Perinatal Mouse Eye

The loss of sight affects approximately 3.4 million people in the United States and is expected to increase in the upcoming years.1 Recently, gene therapy and stem cell transplantations have become key therapeutic tools for treating blindness resulting from retinal degenerative diseases. Several forms of autologous transplantation for age-related macular degeneration (AMD), such as iris pigment epithelial cell transplantation, have generated encouraging results, and human clinical trials have begun for other forms of gene and stem cell therapies.2 These include RPE65 gene replacement therapy in patients with Leber's congenital amaurosis and an RPE cell transplantation using human embryonic stem (ES) cells in Stargardt's disease.3-4 Now that there are gene therapy vectors and stem cells available for treating patients with retinal diseases, it is important to verify these potential therapies in animal models before applying them in human studies. The mouse has become an important scientific model for testing the therapeutic efficacy of gene therapy vectors and stem cell transplantation in the eye.5-8 In this video article, we present a technique to inject gene therapy vectors or stem cells into the subretinal space of the mouse eye while minimizing damage to the surrounding tissue.

This surgical technique illustrates the injection of gene therapy vectors and stem cells into the subretinal space of the mouse eye.

Fare Göz içine Gen Terapisi Vektörler ve Kök Hücre Subretinal Enjeksiyon

The loss of sight affects approximately 3.4 million people in the United States and is expected to increase in the upcoming years.1 Recently, gene therapy ...

Tıp

Retinal Detachment Model in Rodents by Subretinal Injection of Sodium Hyaluronate

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the occurrence of subretinal hemorrhage can affect photoreceptor cell death in the detached retina. Hence, it is advantageous to create reproducible RDs without subretinal hemorrhage for evaluating photoreceptor cell death. We modified a previously reported method to create bullous and persistent RDs in a reproducible location with rare occurrence of subretinal hemorrhage. The critical step of this modified method is the creation of a self-sealing scleral incision, which can prevent leakage of sodium hyaluronate after injection into the subretinal space. To make the self-sealing scleral incision, a scleral tunnel is created, followed by scleral penetration into the choroid with a 30 G needle. Although choroidal hemorrhage may occur during this step, astriction with a surgical spear reduces the rate of choroidal hemorrhage. This method allows a more reproducible and reliable model of photoreceptor death in diseases that involve RD such as rhegmatogenous RD, retinopathy of prematurity, diabetic retinopathy, central serous chorioretinopathy, and age-related macular degeneration (AMD).

Creating experimental retinal detachments with a reproducible and sustained height of detachment, and without subretinal hemorrhage, is important for studying the pathophysiology of photoreceptor cell loss in retinal disease and evaluating potential therapeutic interventions. Here, we report such a method in detail.

Sodyum Hyaluronat subretinal Enjeksiyon Kemirgenler Retina Dekolmanı Modeli

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the ...

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.

Heterotopik Kalp Nakli bir Fare Modeli Kullanarak Pulmoner Vana Nakli

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

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.

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 ...

Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment epithelium (RPE) cells. RPE provide critical support for photoreceptors and death or dysfunction of RPE cells is characteristic of age-related macular degeneration (AMD), the leading cause of permanent vision loss in people age 55 and older. While no cure for AMD has been identified, implantation of healthy RPE in diseased eyes may prove to be an effective treatment, and large numbers of RPE cells can be readily generated from pluripotent stem cells. Several interesting questions regarding the safety and efficacy of RPE cell delivery can still be examined in animal models, and well-accepted protocols used to inject RPE have been developed. The technique described here has been used by multiple groups in various studies and involves first creating a hole in the eye with a sharp needle. Then a syringe with a blunt needle loaded with cells is inserted through the hole and passed through the vitreous until it gently touches the RPE. Using this injection method, which is relatively simple and requires minimal equipment, we achieve consistent and efficient integration of stem cell-derived RPE cells in between the host RPE that prevents significant amount of photoreceptor degeneration in animal models. While not part of the actual protocol, we also describe how to determine the extent of the trauma induced by the injection, and how to verify that the cells were injected into the subretinal space using in vivo imaging modalities. Finally, the use of this protocol is not limited to RPE cells; it may be used to inject any compound or cell into the subretinal space.

Here we present a community accepted protocol in multimedia format for subretinally injecting a bolus of RPE cells in rats and mice. This approach can be used for determining rescue potentials, safety profiles, and survival capacities of grafted RPE cells upon implantation in animal models of retinal degeneration.

Süspansiyon Retina pigment epiteli Hücreleri sunun Kemirgenler Subretinal Enjeksiyonları Sahne

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular weight metabolites in tissues of the treated host to achieve long-term survival. The semipermeable membrane allows engrafted encapsulated cells to avoid rejection by the immune system. The encapsulation procedure was designed to enable a controlled release of bioactive compounds, such as insulin, other hormones, and cytokines. Here we describe a method for encapsulation of catabolic cells, which consume lipids for heat production and energy dissipation (thermogenesis) in the intra-abdominal adipose tissue of obese mice. Encapsulation of thermogenic catabolic cells may be potentially applicable to the prevention and treatment of obesity and type 2 diabetes. Another potential application of catabolic cells may include detoxification from alcohols or other toxic metabolites and environmental pollutants.

Here, we present a protocol for encapsulation of catabolic cells, which consume lipids for heat production in intra-abdominal adipose tissue and increase energy dissipation in obese mice.

Yağ Dokusu Depolarında içine Nakli Enkapsülasyon termojenik Preadipositler

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

Orthotopic Hind Limb Transplantation in the Mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and translational research in the field of transplantation medicine. A vast array of reagents is available exclusively in this setting, including mono- and polyclonal antibodies for both diagnostic and interventional applications. In addition, a vast number of genotyped, inbred, transgenic, and knock out strains allow detailed investigation of the individual contributions of humoral and cellular components to the complex interplay of an immune response and make the mouse the gold standard for immunological research. 
Vascularized Composite Allotransplantation (VCA) delineates a novel field of transplantation using allografts to replace "like with like" in patients suffering traumatic or congenital tissue loss. This surgical methodological protocol shows the use of a non-suture cuff technique for super-microvascular anastomosis in an orthotopic mouse hind limb transplantation model. The model specifically allows for comparison between established paradigms in solid organ transplantation with a novel form of transplants consisting of various different tissue components. Uniquely, this model allows for the transplantation of a viable vascularized bone marrow compartment and niche that have the potential to exert a beneficial effect on the balance of immune acceptance and rejection. This technique provides a tool to investigate alloantigen recognition and allograft rejection and acceptance, as well as enables the pursuit of functional nerve regeneration studies to further advance this novel field of transplantation.

This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in&#160;vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA).

Fare Ortotopik Hind Ekstremite nakli

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

A Step by Step Protocol for Subretinal Surgery in Rabbits

Age related macular degeneration (AMD), retinitis pigmentosa, and other RPE related diseases are the most common causes for irreversible loss of vision in adults in industrially developed countries. RPE transplantation appears to be a promising therapy, as it may replace dysfunctional RPE, restore its function, and thereby vision.
Here we describe a method for transplanting a cultured RPE monolayer on a scaffold into the subretinal space (SRS) of rabbits. After vitrectomy xenotransplants were delivered into the SRS using a custom made shooter consisting of a 20-gauge metallic nozzle with a polytetrafluoroethylene (PTFE) coated plunger. The current technique evolved in over 150 rabbit surgeries over 6 years. Post-operative follow-up can be obtained using non-invasive and repetitive in vivo imaging such as spectral domain optical coherence tomography (SD-OCT) followed by perfusion-fixed histology.
The method has well-defined steps for easy learning and high success rate. Rabbits are considered a large eye animal model useful in preclinical studies for clinical translation. In this context rabbits are a cost-efficient and perhaps convenient alternative to other large eye animal models.

Retinal pigment epithelium (RPE) replacement strategies and gene-based therapy are considered for several retinal degenerative conditions. For clinical translation, large eye animal models are required to study surgical techniques applicable in patients. Here we present a rabbit model for subretinal surgery geared towards RPE transplantation, which is versatile and cost-efficient.

Tavşanlarda Subretinal Cerrahi Adım Protokolle Bir Adım

Age related macular degeneration (AMD), retinitis pigmentosa, and other RPE related diseases are the most common causes for irreversible loss of vision in ...

Transplantation Into the Mouse Ovarian Fat Pad

Orthotopic transplantation assays in mice are invaluable for studies of cell regeneration and neoplastic transformation. Common approaches for orthotopic transplantation of ovarian surface and tubal epithelia include intraperitoneal and intrabursal administration of cells. The respective limitations of these methods include poorly defined location of injected cells and limited space volume. Furthermore, they are poorly suited for long-term structural preservation of transplanted organs. To address these challenges, we have developed an alternative approach, which is based on the introduction of cells and tissue fragments into the mouse fat pad. The mouse ovarian fat pad is located in the immediate vicinity of the ovary and uterine tube (aka oviduct, fallopian tube), and provides a familiar microenvironment for cells and tissues of these organs. In our approach fluorescence-labeled mouse and human cells, and fragments of the uterine tube are engrafted by using minimally traumatic dorsal incision surgery. Transplanted cells and their outgrowths are easily located in the ovarian fat pad for over 40 days. Long-term transplantation of the entire uterine tube allows correct preservation of all principle tissue components, and does not result in adverse side effects, such as fibrosis and inflammation. Our approach should be uniquely applicable for answering important biological questions such as differentiation, regenerative and neoplastic potential of specific cell populations. Furthermore, it should be suitable for studies of microenvironmental factors in normal development and cancer.

We describe an ovarian fad pad transplantation assay suitable for studies of normal and transformed epithelia of the female reproductive tract. The mouse fat pad allows transplantation of large tissue fragments, is easily accessible for surgery and imaging, and provides the most favorable native environment for tissues of M&#252;llerian origin.

Fare Yumurtalık Yağ Pad içine Transplantasyon

Orthotopic transplantation assays in mice are invaluable for studies of cell regeneration and neoplastic transformation. Common approaches for orthotopic ...

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the established technique for a modified blood circuit reconstruction in a heterotopic abdominal heart transplantation model is presented. This method uses the intrathoracic inferior vena cava (IIVC) instead of the pulmonary artery of the donor heart for the anastomosis to the inferior vena cava of the recipient. It is facilitating and improving success rates for abdominal heart transplantation in mice.

A novel technique for blood circuit reconstruction in a heterotopic abdominal mouse heart transplantation model is demonstrated.

Abdominal Fare Kalp Nakli Modelinde Kan Devresi Rekonstrüksiyonu

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the ...