Name: fabrication of tongue extracellular matrix and reconstitution of tongue squamous cell carcinoma in vitro
Uploaded: 2018-06-20T12:30:00
Description: Watch this Scientific Journal Video about Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro at JoVE.com

Yazarlar, Ulusal Doğa Bilimleri Vakfı Çin (31371390), devlet yüksek teknoloji geliştirme projesi (2014AA020702) programı ve programın Guangdong bilim ve teknoloji (2016B030231001) araştırma hibe destek kabul etmiş oluyorsunuz.

Tüm hayvan iş hayvan refahı Yasası, kurumsal yönergelere uygun olarak gerçekleştirilen ve kurumsal hayvan bakım ve kullanım Komitesi, Sun Yat-Sen'in Üniversitesi tarafından onaylanmıştır.
1. TEM hazırlanması
<ol><li>Fareler tarafından servikal çıkığı yürütmek ve steril cerrahi makas ve cımbız kullanarak dilleri kaldırın.</li>
	<li>Tongues yılında % 75 etanol 3 min için bırakın, sonra her dil bir 1,5 mL koymak Eppendorf (EP) tüp ile 1 mL 10 mM steril fosfat t...

<ol>
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Oncol. 29, 1488-1494 (2011).</li><li>Zhao, L., Huang, L., Yu, S., Zheng, J., Wang, H., Zhang, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decellularized+tongue+tissue+as+an+in+vitro.+model+for+studying+tongue+cancer+and+tongue+regeneration.">Decellularized tongue tissue as an in vitro. model for studying tongue cancer and tongue regeneration.</a> Acta Biomaterialia. 58, 122-135 (2017).</li><li>Ng, S. L., Narayanan, K., Gao, S., Wan, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lineage+restricted+progenitors+for+the+repopulation+of+decellularized+heart.">Lineage restricted progenitors for the repopulation of decellularized heart.</a> Biomaterials. 32, 7571-7580 (2011).</li><li>Ott, H. 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B., 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=Initial+binding+and+recellularization+of+decellularized+mouse+lung+scaffolds+with+bone+marrow-derived+mesenchymal+stromal+cells.">Initial binding and recellularization of decellularized mouse lung scaffolds with bone marrow-derived mesenchymal stromal cells.</a> Tissue Eng. Pt A. 18, 1-16 (2012).</li><li>Ott, H. 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=Regeneration+and+orthotopic+transplantation+of+a+bioartificial+lung.">Regeneration and orthotopic transplantation of a bioartificial lung.</a> Nat. Med. 16, 927-933 (2010).</li><li>Petersen, T. 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W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perspectives+on+whole-organ+assembly:+moving+toward+transplantation+on+demand.">Perspectives on whole-organ assembly: moving toward transplantation on demand.</a> J. Clin. Invest. 122, 3817-3823 (2012).</li><li>Song, J. J., Ott, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organ+engineering+based+on+decellularized+matrix+scaffolds.">Organ engineering based on decellularized matrix scaffolds.</a> Trends Mol. Med. 17, 424-432 (2011).</li><li>Badylak, S. 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Decellularized ECM imalat için köklü bir protokol dokularda hücresel bileşenleri kaldırma sırasında yerel ECM kompozisyon tutmak gerekir neredeyse tamamen21. Perfüzyon konvektif taşıma ile hücresel malzemeleri kaldırmak için damarlara aracılığıyla gerektiren şu anda bildirilen decellularization protokolleri rağmen mekanik ajitasyon burada, geleneksel basit ve ucuz Yöntem22 bilinen kabul edilmiştir , 23 , <s...

Dil deglutition, eklem ve tatma gibi çeşitli önemli işlevleri vardır. Böylece, dil işlev bozukluğu hastaların yaşam kalitesi1üzerinde büyük etkisi vardır. Oral kavite içinde en yaygın malignite dil Skuamöz Hücre alkol veya sigara tütün2insan genellikle oluşan Karsinomu (TSCC), olduğunu.
Son yıllarda, TSCC üzerinde temel araştırma az bir gelişme sağlanmıştır. Verimli vitro araştırma eksikliği en büyük sorunlardan biri kalır modelleri. Böylece, hücre dışı matriks (ECM) olası bir çözüm olarak ortaya çıktı. ECM son derece organize matris bileşenlerden oluşan bir karmaşık ağ çerçeve olduğu bir ECM benzeri yapıda ve kompozisyon sahip İskele malzemeleri kanser araştırmaları için yetkili gerekir. Decellularized ECM mükemmel niş için hangi ECM en önemli avantajı için çıkıyor aynı kökenli vitro, hücrelerden sağlar.
ECM doku deterjanlar ve enzimler kullanarak decellularization aracılığıyla gelen kaldırılmakta olan hücresel bileşenleri ile muhafaza edilebilir. Kolajen, fibronektin ve laminin decellularized matris içinde de dahil olmak üzere çeşitli ECM bileşenleri yerel doku gibi microenvironment kültürlü hücreler için hayatta kalma, yayılması ve hücreleri3farklılaşma teşvik sağlar. Ayrıca, ECM hücresel bileşenleri yokluğu ile en az düzeye immünojenisite nakli için azaltılabilir.
Şimdiye kadar decellularized ECM imalat yöntemleri farklı doku ve organlarında, kalp4,5,6,7, karaciğer8,gibi9,10 denendi ,11, akciğer12,13,14,15,16,17ve böbrek18,19 , 20. ancak, ilgili hiçbir araştırma olmak buldu en iyi bildiğimiz kadarıyla dilinde benzer iş.
Bu çalışmada, decellularized dil hücre dışı matriks (TEM) verimli ve uygun fiyatla rezervasyon yaptırın fiziksel, kimyasal ve enzimatik tedavi bir dizi tarafından fabrikasyon. Sonra TEM TSCC vitro, özetlemek için TSCC davranış ve Kalkınma için uygun bir simülasyon gösterilen kullanıldı. TEM iyi biyouyumluluk hücreleri TEM TSCC araştırma3' te büyük potansiyeli olabilir gösterir doku özel niş Kılavuzu yeteneği vardır. Burada gösterilen iletişim kuralı Patogenez ya da TSCC klinik tedaviler üzerinde eğitim araştırmacılar için bir seçim sağlar.

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		<tr height="42">
			<td height="42" style="height:42px;width:216px;">C57-BL/6J mice</td>
			<td style="width:292px;">Sun Yat-sen University Laboratory Animal Center</td>
			<td style="width:119px;"></td>
			<td style="width:264px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ethanol</td>
			<td style="width:292px;">Guangzhou Chemical Reagent Factory</td>
			<td style="width:119px;">HB15-GR-2.5L</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sodium chloride</td>
			<td style="width:292px;">Sangon Biotech</td>
			<td style="width:119px;">A501218</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Potassium chloride</td>
			<td style="width:292px;">Sangon Biotech</td>
			<td style="width:119px;">A100395</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Dibasic Sodium Phosphate</td>
			<td style="width:292px;">Guangzhou Chemical Reagent Factory</td>
			<td style="width:119px;">BE14-GR-500G</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Potassium Phosphate Monobasic&#160;</td>
			<td style="width:292px;">Sangon Biotech</td>
			<td style="width:119px;">A501211</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">1.5 mL EP tube</td>
			<td style="width:292px;">Axygen</td>
			<td style="width:119px;">MCT-150-A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ultra-low temperature freezer&#160;</td>
			<td style="width:292px;">Thermo Fisher Scientific</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">3.5 cm cell culture dish</td>
			<td style="width:292px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">153066</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">6 cm cell culture dish</td>
			<td style="width:292px;">Greiner</td>
			<td style="width:119px;">628160</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Triton X-100</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">V900502</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Calcium chloride</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">746495</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Magnesium chloride</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">449164</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">DNase</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">D5025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Magnesium sulphate</td>
			<td style="width:292px;">Sangon Biotech</td>
			<td style="width:119px;">A601988</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Glucose</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">158968</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sodium bicarbonate</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">S5761</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ampicillin</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">A9393</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Kanamycin</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">PHR1487</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Surgical suture</td>
			<td style="width:292px;">Shanghai Jinhuan</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">250 mL wide-mouth bottle</td>
			<td style="width:292px;">SHUNIU</td>
			<td style="width:119px;">1407</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Magnetic stirrer</td>
			<td style="width:292px;">AS ONE</td>
			<td style="width:119px;">1-4602-32</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">CO2 incubator</td>
			<td style="width:292px;">SHEL LAB</td>
			<td style="width:119px;">SCO5A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">10 mL syringe</td>
			<td style="width:292px;">Hunan Pingan</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">50 mL centrifuge tube</td>
			<td style="width:292px;">Greiner</td>
			<td style="width:119px;">227270</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Cal27 cell</td>
			<td style="width:292px;">Chinese Academy of Science, Shanghai Cell Bank</td>
			<td style="width:119px;"></td>
			<td>Tongue squamous cell carcinoma cell line</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">U2OS cell</td>
			<td style="width:292px;">Chinese Academy of Science, Shanghai Cell Bank</td>
			<td style="width:119px;"></td>
			<td>Human osteosarcoma cell line</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">DMEM/F12</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">D0547</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sodium pyruvate</td>
			<td style="width:292px;">Sigma-Aldrich</td>
			<td style="width:119px;">P5280</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hepes free acid</td>
			<td style="width:292px;">BBI</td>
			<td style="width:119px;">A600264</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">FBS</td>
			<td style="width:292px;">Hyclone</td>
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			<td height="21" style="height:21px;width:216px;">4 &#176;C fridge</td>
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fabrication of tongue extracellular matrix and reconstitution of tongue squamous cell carcinoma in vitro

Dil Skuamöz Hücre Karsinomu (TSCC) vitroiçin etkili ve gerçekçi bir model oluşturmak için yöntemleri TSCC İnşaat için fonksiyonel iskele sağlayan decellularized üretmek dil hücre dışı matriks (TEM) oluşturulmuştur. TEM vitro niş hücre büyümesi, farklılaşma ve hücre geçiş sağlar. Yerel hücre dışı matriks (ECM) ve biyokimyasal besteleri decellularized matris içinde muhafaza microstructures doku özel niş hücreleri demirleme için sağlar. TEM imalatı deoksiribonükleaz (DNaz) sindirim bir ciddi organik veya inorganik Önarıtma ile eşlik tarafından gerçekleştirilebilir. Bu iletişim kuralı kullanımı kolay ve decellularization için yüksek verimlilik sağlar. TEM inşaat TSCC modeli sağlayan olumlu cytocompatibility TSCC hücrelerin çalkalanmış mı statik kültür koşullarda gösterdi. Kendini yetiştirmiş bir biyoreaktör Ayrıca hücre kültürü için kalıcı karıştırılmış koşul için kullanıldı. Yeniden oluşturulan TSCC TEM kullanarak özellikleri ve klinik TSCC histopatoloji, benzer özellikleri TSCC araştırma potansiyeli düşündüren gösterdi.

Bu iletişim kuralı TEM hazırlanması için verimli ve uygun olduğu ortaya çıkıyor. TEM anadili kurcalayarak kıyasla mükemmel decellularization gösterdi. Decellularization etkinliğini hematoksilen-(Şekil 1A-B) boyama eozin tarafından (o) doğrulandı. Sonuçları boyama o nükleer TEM (Şekil 1B) boyama tam ortadan kalkması saptandı. Ayrıca, önceki işten DNA içerik miktar DNA TE...

Watch this Scientific Journal Video about Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro at JoVE.com

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro

Dil hücre dışı matriks ve dil Skuamöz Hücre Karsinomu Vitro sulandırma imalatı

Bir yöntem etkili decellularization ile dil hücre dışı matriks (TEM) hazırlanması için burada gösterilir. TEM fonksiyonel iskele dil Skuamöz Hücre Karsinomu (TSCC) manken çalkalanmış mı statik kültür koşullar altında yeniden inşası için kullanılabilir.

In order to construct an effective and realistic model for tongue squamous cell carcinoma (TSCC) in vitro, the methods were created to produce ...

Research

JoVE Journal

Cancer Research

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro

A Model for Perineural Invasion in Head and Neck Squamous Cell Carcinoma

Baş ve Boyun Skuamöz Hücre Karsinomu içinde perinöral invazyon İçin Bir Model

Perineural invasion (PNI) is found in approximately 40% of head and neck squamous cell carcinomas (HNSCC). Despite multimodal treatment with surgery, ...

A Syngeneic Mouse Model of Metastatic Renal Cell Carcinoma for Quantitative and Longitudinal Assessment of Preclinical Therapies

Preklinik Tedaviler Nicel ve Boyuna Değerlendirme A sinjeneik Fare Modeli Yapmış Bir Renal Hücreli Karsinom

Renal cell carcinoma (RCC) affects &#62; 60,000 people in the United States annually, and ~ 30% of RCC patients have multiple metastases at the time of ...

Four-color Fluorescence Immunohistochemistry of T-cell Subpopulations in Archival Formalin-fixed, Paraffin-embedded Human Oropharyngeal Squamous Cell Carcinoma Samples

Arşiv Formalin Sabitlenmiş, Parafin Gömülü İnsan Orofaringeal Skuamöz Hücreli Karsinoma Örneklerinde T-hücresi Subpopülasyonlarının Dört Renkli Floresan İmmunohistokimyası

The four-color fluorescence immunohistochemistry (IHC) technique is a method to quantify cell populations of interest while taking into account their ...

Moving Upwards: A Simple and Flexible In Vitro Three-dimensional Invasion Assay Protocol

Moving Upwards: A Simple and Flexible In Vitro Three-dimensional Invasion Assay Protocol

Yukarı doğru hareket: Bir basit ve esnek Vitro üç boyutlu işgali tahlil iletişim kuralı

Although 3D invasion assays have been developed, the challenge remains to study cells without affecting the integrity of their microenvironment. ...

Analyzing the Communication Between Monocytes and Primary Breast Cancer Cells in an Extracellular Matrix Extract (ECME)-based Three-dimensional System

Analyzing the Communication Between Monocytes and Primary Breast Cancer Cells in an Extracellular Matrix Extract ECME-based Three-dimensional System

Monosit ve bir hücre dışı matriks içinde birincil meme kanseri hücreleri arasındaki iletişimi analiz özü (ECME)-üç boyutlu sistem tabanlı

Embedded in the extracellular matrix (ECM), normal and neoplastic epithelial cells intimately communicate with hematopoietic and non-hematopoietic cells, ...

Therapy Testing in a Spheroid-based 3D Cell Culture Model for Head and Neck Squamous Cell Carcinoma

Baş ve boyun Skuamöz hücreli karsinom için bir küresel tabanlı 3D hücre kültür modelinde test terapi

Current treatment options for advanced and recurrent head and neck squamous cell carcinoma (HNSCC) enclose radiation and chemo-radiation approaches with ...

Intramucosal Inoculation of Squamous Cell Carcinoma Cells in Mice for Tumor Immune Profiling and Treatment Response Assessment

Skuamöz Hücre Karsinomu Hücre Tümörü bağışıklık profil oluşturma için fareler ve tedaviye yanıt değerlendirme intramucosal aşı

Head and neck squamous cell carcinoma (HNSCC) is a debilitating and deadly disease with a high prevalence of recurrence and treatment failure. To develop ...

Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma

Üç boyutlu kemik hücre dışı matriks modeli osteosarkom için

Osteosarcoma (OS) is the most common and a highly aggressive primary bone tumor. It is characterized with anatomic and histologic variations along with ...

Studying Normal Tissue Radiation Effects using Extracellular Matrix Hydrogels

Ekstrasellüler matris Hidrogeller kullanarak normal doku radyasyon etkileri eğitim

Radiation is a therapy for patients with triple negative breast cancer. The effect of radiation on the extracellular matrix (ECM) of healthy breast tissue ...

Time-Lapse 2D Imaging of Phagocytic Activity in M1 Macrophage-4T1 Mouse Mammary Carcinoma Cells in Co-cultures

Eş-Kültürlerde M1 Makrofaj-4T1 Fare Mammary Karsinom Hücrelerinde Fagositik Aktivitenin Zaman Atlamalı 2D Görüntülemesi

Tumor-associated macrophages (TAMs) have been identified as an important component for tumor growth, invasion, metastasis, and resistance to cancer ...