Yazarlar hiçbir katkıda bulunanlar var.

Protokol Üniversitesi Hastanesi Zürih insan araştırma Etik Komitesi kuralları izler.
1. Tanı Neuromata ve hasta seçimi
<ol><li>Ameliyat öncesi sinir blokları için nöroma proksimal tanılar onaylamak için gerçekleştirin. Önkol orta üçüncü bloklarını değerlendirmek ve SBRN, lateral antebrachial Kutanöz sinir (LACN), posterior interosseöz sinir (PIN) ve ulnar sinir dorsal bir dalı gibi örtüşen innervasyon alanlarında potansiyel Tetikleyicileri hariç 10.</li>
	<li>Sadece acı azaltma (en az 5 Puan ağrı için görsel Analog ölçek (VAS) üzerinde)11olan hastalarda ameliyat.</li>
</ol>2. cerrahi hazırlık
<ol><li>Hastanın herhangi bir yiyecek veya sıvı alımı ameliyat öncesi 6 h yasaklıyorum.</li>
	<li>Hastanın cerrahi suite ortasında evrensel operatif tablodaki yinelenen bir plaj sandalye konuma yerleştirin.</li>
	<li>Etkilenen kol/el kol dayanağı koyun.</li>
	<li>Endotrakeal anestezi gerçekleştirmek için Anestezistler sormak (µg/kg fentanil 2, rocuronium 0.6 mg/kg, propofol 150-250 mg).</li>
	<li>Cilt povidone iyot alan liposuction (örneğin iç uyluk ve/veya üst/alt karın) ve site nöroma eksizyonu ile dezenfekte.</li>
	<li>Hasta steril cerrahi perdeler ile kapağı, liposuction ve cerrahi siteleri maruz bırakarak.</li>
</ol>3. şişen Liposuction
<ol><li>#11-bıçak neşter ile yaklaşık 3-5 mm çapında birkaç küçük bıçak (delik) insizyon liposuction alanında olun. Subkutan katman ulaşılana kadar deşmek.</li>
	<li>İçinde bir seyreltik lidokain ve epinefrin (500 mL zil'ın laktat çözüm, 20 mL %1 lidokain ve 1 mg epinefrin) yolu ile (kanül infiltre) 14'lük yaklaşıyor subkutan yağ katmanına (3 mm derinlik) içeren şişen çözüm enjekte Bölge şişman hasat.</li>
	<li>Şişen çözüm etkili olması için izin en az 20 dakika bekleyin.</li>
	<li>Kanca 3 mm toplama kanül (dış çapı 3 mm, boyu 26 cm) 60 mL Toomey şırınga ve yağ bıçak insizyon ile hasat Başlat. Not: Nondominant el sürekli yerleşim ve tabii ki kanül denetler.</li>
	<li>60 mL işaretine Toomey şırınga pistonu geri çekin ve bir sivrisinek kelepçe şırınga negatif basınç ve dominant el tarafından sürekli bir vakum oluşturma önlemek için içine kapanık pistonu boyunca yerleştirin.</li>
	<li>220 mL yağ dokusunun en az sağlanan Toomey şırınga stand dikey konumda Toomey şırınga konumlandırma tarafından hasat; Bu yağ ve sıvı ayrılması sedimantasyon sağlar.</li>
	<li>Kaliteli yağ greft önemli bir miktar kan olmadan elde etmek için özel dikkat. Not: bölgeyi liposuction artar kan içerik farketmesinden değiştirin.</li>
	<li>Bıçak insizyon ile basit Dikiş dikiş ve baskı uygulamak gerektiği gibi (örneğin basınç kuşağı).</li>
	<li>Lipoaspirate tek kullanımlık steril tek kullanımlık birtakım yalıtım sistemi aktarın.</li>
	<li>Bir Toomey şırınga lipoaspirate ile sağlanan tribünde daha sonra lipofilling için yağ kesir elde etmek için devam et.</li>
</ol>4. yağ doku işleme
<ol><li>Steril tüketim kümesini kapatın. Not: Yalıtım sistemi ( Tablo malzemelerigörmek) tam ıslak bir test yaparak onaylar.</li>
	<li>Ekrandaki yönergeleri izleyin. Not: Çoğunlukla yazılım kontrollü ve yalnızca el ile onay için listelenen adımları "sonraki" veya "geri" kontrol paneli üzerindeki düğmelere basarak gerektirir işlemidir. Lipoaspirate doku toplama tüp aktarılır. Doku toplama tüp lipoaspirate tartılır ve otomatik olarak kalan kan hücreleri ve ıslatma çözüm kaldırmak için zil'ın laktat çözüm ile temizlenir.</li>
	<li>Daha fazla yağ gerekirse ekleyin. Not: hasat yağ doku ağırlığına bağlı olarak yalıtım sistemi daha fazla yağ eklemek için kullanıcıdan veya eklemek için ek enzim reaktif miktarını hesaplar.</li>
	<li>Yalıtım sistem tarafından istendiğinde enzim reaktif gerekli miktarda ekleyin. Not: Enzimatik sindirim işlemi sırasında lipid ve SVF ayrılması sağlanır. Böylece, hareket salınan dokudur. Enzimatik sindirim tamamlandıktan sonra SVF hücre yoğunlaşması santrifüj odasına otomatik olarak transfer edilir. Santrifüj odasında SVF hücre Pelet konsantre (santrifüj hızı ise yaklaşık 3000 x g). Son olarak, hücre Pelet yıkar kalan enzim düzeylerinin çıktı azaltmak için zil'ın laktat solüsyonu ile bir dizi uğrar. İşlem tamamlandığında, net sıvı (SVF) 5 mL alınır.</li>
</ol>5. cerrahi müdahale
<ol><li>Nöroma yaklaşım önkol distal üçüncü daha önce teşhisi nöroma site üzerinden bir cilt kesi yapın.</li>
	<li>Cerrahi Binoküler büyüteçler Büyüteç gözlük (2.5-3.5 X) kullanımı altında subkutan doku teşrih ve işin içinde sinir nöroma için proksimal belirlemek devam edin.<ol>
<li>Daha sonra sinir klemple izleyin ve Nörom (görsel muayene on bağlı olarak) tanımlar.</li>
	</ol>
</li>
	<li>Nöroma tam çekiş temizlendikten sonra bir düz trans-nöro makas kesim tarafından nöroma tüketim başlar. Hiçbir nöroma belirlenirse, yalnızca neurolysis sinirin gerçekleştirilir.</li>
	<li>Kalan sinir güdük neurolysis ile 3 cm için bilek proksimal kadar devam edin. Not: Sinir güdük sonuna asıl kesi ortasında yer alan olmalıdır.</li>
	<li>İki ila dört küçük delik (bıçak) insizyon (1 mm çapında) epidermal ve dermal cutis asıl yaklaşım çevresinde aracılığıyla bir neşter ile düzenlemek.</li>
	<li>2-4 künt Kanüller yerleştirin (16 lik uzunluğu 9 cm) güdük için bıçak kesikleri perineural üzerinden. Not: Kanüller ucu sinir güdük doğru yönlendirilmelidir.</li>
	<li>Kanüller yerinde yapıştırıcı soyunma tarafından güvenli.</li>
	<li>Dikiş Kanüller çevresinde yer ve belgili tanımlık suture maddi in situ durumdaki düğüm güvenliğini sağlama olmadan bırakın.</li>
	<li>Dikiş sıkıca aşılama çıkım önlemek için ana kesi. Not: Kanüller in situkalmasını sağlamak.</li>
</ol>6. SVF zenginleştirilmiş yağ grefti uygulanması
Not: işlem yaklaşık 1,5 saat sonra işlenmiş SVF 5 mL daha fazla kullanıma hazırdır.
<ol><li>SVF 5 mL içinde bir 10 mL iletişim şırınga Aspire edin.</li>
	<li>Aynı şekilde, karşı 10 mL şırınga posalı lipid fraksiyonunun 2 mL Aspire edin.</li>
	<li>5 mL SVF de bir şırınga pistonu geri çekilmesi ve yeterli karıştırma elde edilir kadar iletişim, karşıt şırınga pistonu çekerek alternatif tarafından posalı lipid kesir 2 mL ile karıştırın.</li>
	<li>Eşit olarak 2-4 10 mL şırınga SVF zenginleştirilmiş yağ greft bölün.</li>
	<li>Diğer aynı derecede sinir çevresinde güdük sonra künt Kanüller şırıngaları bağlanarak hazırlanan SVF zenginleştirilmiş yağ dokuları bir dağıtın. Not: greft uygulamadan önce Kanüller ve doğru konumda güvenli Kanüller değil dislocate yağ greft uygulanırken dikkatli olun.</li>
	<li>Kanüller dikkatli bir şekilde çıkarın.</li>
	<li>Sonra küçük bir delik insizyon steri-şeritler ile teyp.</li>
	<li>El ateli içsel olarak artı pozisyon (, 60-70 ° fleksiyondayken metakarpofalangeal (MCP) eklem, tamamen uzun interfalangeal (IP) eklem, başparmak yumruk projeksiyon, 25 ° dorsal fleksiyon bilekte) uygulayarak hareketsiz. Sinir neurolysis gerçekleştirilen yalnızca, hiçbir ateli gereklidir ve kol ve bilek için 10 gün sargılı.</li>
</ol>7. ameliyat sonrası hasta taşıma
<ol><li>Hasta için ameliyat sonrası sonrası anestezi kurtarma birimi aktarmak ve yeterli koğuşunda izlemek için izin verir.</li>
	<li>Ameliyattan sonra 10 gün boyunca bilek hareketsiz devam etmek.</li>
	<li>48 saat sonra ameliyat sonrası bandaj ilk değişiklik yapabilir ve (Yani her iki günde kadar deşarj) düzenli aralıklarla devam.</li>
	<li>Dikişler 12-14 gün postoperatively kaldırın.</li>
	<li>2, 6, 12, takip sırasında ağrı ve 36 ay klinik değerlendirmesi gerçekleştir.</li>
</ol>

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Yazarlar ifşa gerek yok.

Bu da çalışmanın amacı yöntemli semptomatik sonu-neuromata için yeni bir tedavi olarak yağ grefti yapılması SVF zenginleştirilmiş teknik yönlerini göstermektir.
SBRN neuromata cerrahi başarı oranı sadece % 3312ile ilişkilidir. Vaienti ve ark. sunulan bir dizi olumlu sonuçları erken takip acı neuromata üst ekstremite58 hastalarda SVF eklemeden perineural (PFG) yağ aşılama kullanar...

Semptomatik sonu-neuromata SBRN yönetimi çok sayıda mevcut cerrahi tedaviler ile zorlu kalır. Prensip olarak, nöroma varlığına ve kalan sinir güdük kasları1,2, kemik3ya da damarlar4gibi komşu yapıların içine taşındı. Bu Yenileyici sinirsel fascicles uç organ havasını taklit ederek dışarı çimlenme önlemek için kabul edilir. Ancak, hiçbir bu tekniklerin üstün, ağrı nüks ve yetersiz acı azaltma yüksek oranda kez gözlendi gibi değildi.
Son zamanlarda, yağ kalan sinir güdük aşılama umut verici sonuçlar vermiştir ve bir yeni tedavi seçeneği üst ekstremite5için acı sonu-neuromata savundu. Sürekli hareket önlemek ve ağrı dokunun mekanik bir engel olarak hareket yanı sıra yağ aşılama faydalı yönü oldukça hücresel düzeyde atfedilir. İnsan yağ dokusu preadipocytes, mast hücreleri, perisitlerden, endotel hücreleri, vasküler düz kas hücreleri, fibroblastlar, dahil olmak üzere birden çok hücre tipleri içerir ve yağ kök hücreleri (ADSCs) elde edilen hematopoetik kök hücre6ile birlikte. Bu türdeş olmayan hücre nüfus Adipose-Derived yenilenme hücreleri (ADRCs) veya SVF denir ve lipoaspirate bir yalıtım sistemi tarafından elde edilebilir ( Tablo malzemelerigörmek) ve uygulamalı enzim reaktif (bkz: malzemeler tablo )7. Tam etki mekanizması hala tam anlaşılmaktadır; Ancak, SVF anjiogenezi, immunomodulation, farklılaşma ve hücre dışı matriks8salgılanmasını iyileştirmeler ile ilişkili olduğu düşünülmektedir.
Son yıllarda yapılan çalışmalarda ağrı nüks yağ eklenen SVF aşılama ile 12 ay sonra ortaya SVF konsantre hücre nüfusu rezorpsiyonu oranını azaltmak için ve yağ dokuları yeniden üretim potansiyelini artırmak amacı ile yağ greft eklenir 5. Ayrıca, aynı zamanda sinir kendisi üzerinde yararlı etkiler sağlar biyolojik eylem dağınık aksonlar, güdük9 ve sinir sıkışması neden olabilir sonraki yara adherences çimlenme azaltarak.
SBRN semptomatik sonu-neuromata tedavisi için yeni bir cerrahi seçenek olduğu gibi SVF zenginleştirilmiş yağ aşılama açıklamak için ilk klinik çalışmada son zamanlarda10yayınlandı. Genel olarak, teknik yanlışlıkla veya iyatrojenik sinir yaralanması sonra meydana gelen herhangi bir nöroma uygulanabilir. Sonraki çalışmanın sistemli bir şekilde bu yordamı teknik yönlerini göstermek için amaçtır.

<table border="0" cellpadding="0" cellspacing="0" width="688">
	<tbody>
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			<td height="21" style="height:21px;width:331px;">Cytori Celution System</td>
			<td style="width:71px;">Cytori</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">isolation system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Celase</td>
			<td style="width:71px;">Cytori</td>
			<td style="width:119px;"></td>
			<td>enyzme reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">liposuction system</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:331px;">tumescent solution (500 ml Lactated Ringers, 20 ml of 1% lidocaine and 1 mg epinephrine)</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">14-gauge infiltrator</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3 mm-collecting cannula (15, 26, 32 cm)</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">60 mL Tommey syringes</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">mosquito clamp</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Toomey syringe stand</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:331px;">4 cannulas (fat injecting cannulas 1.5 mm, syringe NO.2, 90 mm)</td>
			<td style="width:71px;">Chalybs</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">10 mL syringes</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">connecting device for syringes</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">#11- and #15-blade scalpel</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">stevens scissors</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">pins pincette</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">bipolar electrocautery</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">suture material</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">adhesive dressing</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">binocular loupes magnifier glasses (2.5-3.5x)</td>
			<td style="width:71px;">Carl Zeiss</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:331px;">Statistical Package for Social Sciences, version 22</td>
			<td>IBM</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

stromal vascular fraction-enriched fat grafting for the treatment of symptomatic end-neuromata

Bu çalışmada sistemli bir şekilde göstermek ve stromal vasküler kesir (SVF) çok önemli adımları vurgulamak oldu-şişman semptomatik sonu-neuromata çevre duyusal sinir ve bu çalışmada yeni bir tedavi olarak özellikle aşılama zenginleştirilmiş Radyal sinir (SBRN) Yüzeysel dalı. Mevcut tedaviler çok sayıda rağmen inatçı Postoperatif ağrı ve yaygın ağrı nüks hala çok yaygın, bağımsız olarak değerlendirildi yordamı. Nöroma microsurgically eksize buna göre için standart bir protokoldür. Anatomik yapıları, kas veya kemik gibi komşu Yenileyici sinir güdük tehcir yerine yağ greft uygulanan perifocally ve mekanik bir engel olarak. Şişman rezorpsiyonu oranını azaltmak ve dokuları yeniden üretim potansiyelini arttırmak amacıyla, yüksek konsantrasyonlu SVF aşılama içinde entegre edilmiştir. SVF subcutaneous yağlı belirli ticari yalıtım sistemi tarafından lipoaspirate enzimatik ve mekanik ayrılması tarafından izole edilmiştir. SVF zenginleştirilmiş yağ greft mekanik bir engel ve hücresel düzeyde, anjiogenez, inflamasyon ve fibrozis iyileştirilmesi de dahil olmak üzere çeşitli biyolojik etkileri sağlar. Mekanik ve biyolojik etkileri sinir güdük dağınık aksonal akıbet sinir yeniden oluşturma işlemi sırasında azaltmak ve dolayısıyla acı sonu-neuromata nüks önlemek yardımcı olur.

Veri istatistiksel yazılım kullanarak ve ± standart sapma anlamına gelir olarak sunulan analiz edildi. Kolmogorov-Smirnov sınaması çalışma nüfusunun parametrik olmayan dağıtım doğrulamak için uygulandı ve negatif bir ampirik dağıtım doğruladı. Çalışma nüfus parametrik olmayan niteliğine bağlı olarak, Wilcoxon'ın test ilgili örnekleri için ameliyat öncesi ve ameliyat sonrası sonuçları karşılaştırmak için istihdam edildi. İstatistiksel anlamlılık p...

Watch this Scientific Journal Video about Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata at JoVE.com

Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata

Stromal vasküler kesir zenginleştirilmiş şişman semptomatik sonu-neuromata tedavisi için aşılama

Çalışmada stromal vasküler kesir (SVF) teknik yordamı göstermektir-yağ aşılama semptomatik sonu-neuromata yeni bir tedavi olarak zenginleştirilmiş. Bu tekniğin mekanik bariyer ve işlenmiş ve konsantre SVF tarafından hücresel düzeyde biyolojik eylem avantajları sağlar.

The purpose of this study was to methodically illustrate and highlight the crucial steps of stromal vascular fraction (SVF)-enriched fat grafting as a ...

stromal-vascular-fraction-enriched-fat-grafting-for-treatment

Research

JoVE Journal

Medicine

7.6K Görüntüleme.  University Hospital Zurich. Çalışmada stromal vasküler kesir (SVF) teknik yordamı göstermektir-yağ aşılama semptomatik sonu-neuromata yeni bir tedavi olarak zenginleştirilmiş. Bu tekniğin mekanik bariyer ve işlenmiş ve konsantre SVF tarafından hücresel düzeyde biyolojik eylem avantajları sağlar.

Video: Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata

Experimental Generation of Carcinoma-Associated Fibroblasts (CAFs) from Human Mammary Fibroblasts

Carcinomas are complex tissues comprised of neoplastic cells and a non-cancerous compartment referred to as the 'stroma'. The stroma consists of extracellular matrix (ECM) and a variety of mesenchymal cells, including fibroblasts, myofibroblasts, endothelial cells, pericytes and leukocytes 1-3.
The tumour-associated stroma is responsive to substantial paracrine signals released by neighbouring carcinoma cells. During the disease process, the stroma often becomes populated by carcinoma-associated fibroblasts (CAFs) including large numbers of myofibroblasts. These cells have previously been extracted from many different types of human carcinomas for their in vitro culture. A subpopulation of CAFs is distinguishable through their up-regulation of &alpha;-smooth muscle actin (&alpha;-SMA) expression4,5. These cells are a hallmark of 'activated fibroblasts' that share similar properties with myofibroblasts commonly observed in injured and fibrotic tissues 6. The presence of this myofibroblastic CAF subset is highly related to high-grade malignancies and associated with poor prognoses in patients.
Many laboratories, including our own, have shown that CAFs, when injected with carcinoma cells into immunodeficient mice, are capable of substantially promoting tumourigenesis 7-10. CAFs prepared from carcinoma patients, however, frequently undergo senescence during propagation in culture limiting the extensiveness of their use throughout ongoing experimentation. To overcome this difficulty, we developed a novel technique to experimentally generate immortalised human mammary CAF cell lines (exp-CAFs) from human mammary fibroblasts, using a coimplantation breast tumour xenograft model.
In order to generate exp-CAFs, parental human mammary fibroblasts, obtained from the reduction mammoplasty tissue, were first immortalised with hTERT, the catalytic subunit of the telomerase holoenzyme, and engineered to express GFP and a puromycin resistance gene. These cells were coimplanted with MCF-7 human breast carcinoma cells expressing an activated ras oncogene (MCF-7-ras cells) into a mouse xenograft. After a period of incubation in vivo, the initially injected human mammary fibroblasts were extracted from the tumour xenografts on the basis of their puromycin resistance 11.
We observed that the resident human mammary fibroblasts have differentiated, adopting a myofibroblastic phenotype and acquired tumour-promoting properties during the course of tumour progression. Importantly, these cells, defined as exp-CAFs, closely mimic the tumour-promoting myofibroblastic phenotype of CAFs isolated from breast carcinomas dissected from patients. Our tumour xenograft-derived exp-CAFs therefore provide an effective model to study the biology of CAFs in human breast carcinomas. The described protocol may also be extended for generating and characterising various CAF populations derived from other types of human carcinomas.

Experimental Generation of Carcinoma-Associated Fibroblasts CAFs from Human Mammary Fibroblasts

Carcinoma-associated fibroblasts (CAFs) rich in myofibroblasts present within the tumour stroma, play a major role in driving tumour progression. We developed a coimplantation tumour xengraft model for experimentally generating CAFs from human mammary fibroblasts. The protocol describes how to establish CAF myofibroblasts that acquire an ability to promote tumourigenesis.

İnsan Meme Fibroblastlar Karsinom İlişkili Fibroblastlar Deneysel Nesil (CAFs)

Carcinomas are complex tissues comprised of neoplastic cells and a non-cancerous compartment referred to as the 'stroma'. The stroma consists of ...

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Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

Damar Hastalığı Çalışmaları Farelerde Ortotopik Aort Nakli

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Fare Aortocaval Fistül Teknik Yönleri

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Three-dimensional Co-culture Model for Tumor-stromal Interaction

Cancer progression (initiation, growth, invasion and metastasis) occurs through interactions between malignant cells and the surrounding tumor stromal cells. The tumor microenvironment is comprised of a variety of cell types, such as fibroblasts, immune cells, vascular endothelial cells, pericytes and bone-marrow-derived cells, embedded in the extracellular matrix (ECM). Cancer-associated fibroblasts (CAFs) have a pro-tumorigenic role through the secretion of soluble factors, angiogenesis and ECM remodeling. The experimental models for cancer cell survival, proliferation, migration, and invasion have mostly relied on two-dimensional monocellular and monolayer tissue cultures or Boyden chamber assays. However, these experiments do not precisely reflect the physiological or pathological conditions in a diseased organ. To gain a better understanding of tumor stromal or tumor matrix interactions, multicellular and three-dimensional cultures provide more powerful tools for investigating intercellular communication and ECM-dependent modulation of cancer cell behavior. As a platform for this type of study, we present an experimental model in which cancer cells are cultured on collagen gels embedded with primary cultures of CAFs.

Here we present a protocol to co-culture in three-dimensions, which is useful for investigating multicellular interactions and extracellular matrix-dependent modulation of cancer cell behavior. In this experimental model, cancer cells are cultured on collagen gels embedded with human cancer-associated fibroblasts.

Tümör-stromal Etkileşim için üç boyutlu Co-kültür Modeli

Cancer progression (initiation, growth, invasion and metastasis) occurs through interactions between malignant cells and the surrounding tumor stromal ...

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 ...

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control seizures. However, a significant minority of patients continues to exhibit post-operative seizures, even in those cases in which the suspected source of seizures has been correctly localized and resected. The protocol presented here combines a clinical procedure routinely employed during the pre-operative evaluation of temporal lobe epilepsy (TLE) patients with a novel technique for network analysis. The method allows for the evaluation of the temporal evolution of mesial network parameters. The bilateral insertion of foramen ovale electrodes (FOE) into the ambient cistern simultaneously records electrocortical activity at several mesial areas in the temporal lobe. Furthermore, network methodology applied to the recorded time series tracks the temporal evolution of the mesial networks both interictally and during the seizures. In this way, the presented protocol offers a unique way to visualize and quantify measures that considers the relationships between several mesial areas instead of a single area.

This protocol describes a procedure to track the evolution of mesial network measures in temporal lobe epilepsy (TLE) patients. It is based on the combination of intracranial recordings with a novel numerical technique for data analysis. Specifically, we present a protocol for network analyses of foramen ovale recordings.

İlaca dirençli Temporal Lob Epilepsi Hastalarında Foramen Ovale Elektrot Kayıtları Ağ Analizi

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control ...

Trans-Tympanic Drug Delivery for the Treatment of Ototoxicity

The systemic administration of protective agents to treat drug-induced ototoxicity is limited by the possibility that these protective agents could interfere with the chemotherapeutic efficacy of the primary drugs. This is especially true for the drug cisplatin, whose anticancer actions are attenuated by antioxidants which provide adequate protection against hearing loss. Other current or potential otoprotective agents could pose a similar problem, if administered systemically. The application of various biologicals or protective agents directly to the cochlea would allow for high levels of these agents locally with limited systemic side effects. In this report, we demonstrate a trans-tympanic method of delivery of various drugs or biological reagents to the cochlea, which should enhance basic science research on the cochlea and provide a simple way of directing the use of otoprotective agents in the clinics. This report details a method of trans-tympanic drug delivery and provides examples of how this technique has been used successfully in experimental animals to treat cisplatin ototoxicity.

We present a technique for localized administration of drugs through the trans-tympanic route into the cochlea. Drug delivery through this route would not interfere with the anti-cancer efficacy of the chemotherapeutic drugs such as cisplatin.

Trans-Tympanic ilaç dağıtım ototoksisite tedavisi için

The systemic administration of protective agents to treat drug-induced ototoxicity is limited by the possibility that these protective agents could ...

Clinical Protocol of Producing Adipose Tissue-Derived Stromal Vascular Fraction for Potential Cartilage Regeneration

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the knees by using different forms of mesenchymal stem cells (MSCs). In Korea, bone marrow concentrates and cord blood-derived stem cells have been approved by the Korean Food and Drug Administration (KFDA) for cartilage regeneration. In addition, an adipose tissue-derived stromal vascular fraction (SVF) has been allowed by the KFDA for joint injections in human patients. Autologous adipose tissue-derived SVF contains extracellular matrix (ECM) in addition to mesenchymal stem cells. ECM excretes various cytokines that, along with hyaluronic acid (HA) and platelet-rich plasma (PRP) activated by calcium chloride, may help MSCs to regenerate cartilage and improve knee functions. In this article, we presented a protocol to improve knee functions by regenerating cartilage-like tissue in human patients with OA. The result of the protocol was first reported in 2011 followed by a few additional publications. The protocol involves liposuction to obtain autologous lipoaspirates that are mixed with collagenase. This lipoaspirates-collagenase mixture is then cut and homogenized to remove large fibrous tissue that may clog up the needle during the injection. Afterwards, the mixture is incubated to obtain adipose tissue-derived SVF. The resulting adipose tissue-derived SVF, containing both adipose tissue-derived MSCs and remnants of ECM, is injected into knees of patients, combined with HA and calcium chloride activated PRP. Included are three cases of patients who were treated with our protocol resulting in improvement of knee pain, swelling, and range of motion along with MRI evidence of hyaline cartilage-like tissue.

Here, we present a protocol to produce an adipose tissue-derived stromal vascular fraction and its application to improve knee functions by regenerating cartilage-like tissue in human patients with osteoarthritis.

Yağ dokusundan elde edilen Stromal vasküler kesir için potansiyel kıkırdak rejenerasyon üreten klinik protokol

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the ...

Isolation of Adipose Derived Regenerative Cells for the Treatment of Erectile Dysfunction Following Radical Prostatectomy

Stem cells are used in many research areas within regenerative medicine in part because these treatments can be curative rather than symptomatic. Stem cells can be obtained from different tissues and several methods for isolation have been described. The presented method for the isolation of adipose-derived regenerative cells (ADRCs) can be used within many therapeutic areas because the method is a general procedure and, therefore, not limited to erectile dysfunction (ED) therapy. ED is a common and serious side effect to radical prostatectomy (RP) since ED often is not well treated with conventional therapy. Using ADRC&#8217;s as treatment for ED has attracted great interest due to the initial positive results after a single injection of cells into the corpora cavernosum. The method used for the isolation of ADRC&#8217;s is a simple, automated process, that is reproducible and ensures a uniform product. Furthermore, the sterility of the isolated product is ensured because the entire process takes place in a closed system. It is important to minimize the risk of contamination and infection since the stem cells are used for injection in humans. The whole procedure can be done within 2.5-3.5 hours and does not require a classified laboratory which eliminates the need for shipping tissue to an off-site. However, the procedure has some limitations since the minimum amount of drained lipoaspirate for the isolation device to function is 100 g.

Precise disclosure of methods and protocols is crucial for large scale uptake of stem cell therapies. Here, we present a protocol to isolate adipose-derived regenerative cells, used for a single intracavernous injection as treatment of erectile dysfunction (ED) following radical prostatectomy (RP).

Radikal Prostatektomi Sonrası Erektil Disfonksiyon Tedavisinde Yağ Kaynaklı Rejeneratif Hücrelerin İzolasyonu

Stem cells are used in many research areas within regenerative medicine in part because these treatments can be curative rather than symptomatic. Stem ...

Establishing a Swine Model of Post-myocardial Infarction Heart Failure for Stem Cell Treatment

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major causes of mortality and morbidity around the world. Cell-based therapies for cardiac repair and improvement of left ventricular function after MI have attracted considerable attention. Accordingly, the safety and efficacy of these cell transplantations should be tested in a preclinical large animal model of HF prior to clinical use. Pigs are widely used for cardiovascular disease research due to their similarity to humans in terms of heart size and coronary anatomy. Therefore, we sought to present an effective protocol for the establishment of a porcine chronic HF model using closed-chest coronary balloon occlusion of the left circumflex artery (LCX), followed by rapid ventricular pacing induced with pacemaker implantation. Eight weeks later, the stem cells were administered by intramyocardial injection in the peri-infarct area. Then the infarct size, cell survival, and left ventricular function (including echocardiography, hemodynamic parameters, and electrophysiology) were evaluated. This study helps establish a stable preclinical large animal HF model for stem cell treatment.

We sought to establish a swine model of heart failure induced by left circumflex artery blockage and rapid pacing to test the effect and safety of intramyocardial administration of stem cells for cell-based therapies.

Kök Hücre Tedavisi Için Post-miyokard Enfarktüsü Kalp Yetmezliği Domuz Modeli Oluşturma

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major ...