Name: induction and analysis of oxidative stress in sleeping beauty transposon-transfected human retinal pigment epithelial cells
Uploaded: 2020-12-11T13:00:00
Description: Watch this Scientific Journal Video about Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells at JoVE.com

Yazarlar mükemmel teknik yardım için Gregg Sealy ve Alain Conti'ye ve Berlin'deki Max-Delbrück Merkezi'nden Prof. Zsuzsanna Izsvák'a pSB100X ve pT2-CAGGS-Venus plazmidlerini nazikça sağladıkları için teşekkür eder. Bu çalışma İsviçre Ulusal Bilimler Vakfı ve Avrupa Komisyonu tarafından Yedinci Çerçeve Programı kapsamında desteklendi. Z.I, Avrupa Araştırma Konseyi, ERC Advanced [ERC-2011-ADG 294742] tarafından finanse edildi.

İnsan gözlerinin toplanması ve kullanımına ilişkin prosedürler Kanton Etik Araştırma Komisyonu (no. 2016-01726) tarafından onaylanmıştır.
1. Hücre izolasyonu ve kültür koşulları
<ol><li>İnsan ARPE-19 hücre hattı<ol>
<li>Kültür 5 x 105 ARPE-19 hücreleri, bir insan RPE hücre hattı, Dulbecco'nun Modifiye Kartal Orta / Besin Karışımı F-12 Jambon (DMEM / Ham'ın F-12) % 10 fetal sığır serumu (FBS), 80 U / mL penisilin ile desteklenmiş, 80 μg/mL strept...

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Burada sunulan protokol, herhangi bir putatif faydalı genle transkârlaşmış hücrelere uygulanabilen transfected hücreler tarafından üretilen PEDF ve GM-CSF'nin anti-oksidatif ve koruyucu işlevini analiz etmek için bir yaklaşım sunmaktadır. Genetiği değiştirilmiş hücreleri naklederek proteinleri dokuya ulaştırma amacına sahip gen terapötik stratejilerinde, protein ekspresyon seviyesi, ifadenin uzun ömürlülüğü ve hastalığın bir modelinde ifade edilen proteinin etkinliği hakkında bilgi edinm...

Burada açıklanan model, hücrelerdeki oksidatif stresi azaltmak içinbiyofarmasötik ajanların verimliliğini değerlendirmek için yararlı bir yaklaşım sunar. Modeli, PEDF ve GM-CSF'nin yüksek düzeyde O 2 ve görünür ışığa maruz kalan retina pigment epitel hücreleri üzerindeki H2O2aracılı oksidatif stres üzerindeki koruyucu etkilerini ve fotoreceptör dış segment membranlarının fagositozunu araştırmak için kullandık, önemli düzeyde reaktif oksijen türü (ROS)1, 2. Avasküler yaşa bağlı makula dejenerasyonu (aAMD) 3 , 4 , 5 ,6,7,8patogenezine önemli bir katkıda bulunurlar. Ayrıca, RPE sentezlenmiş nöroprotektif faktörlerde bir azalma vardır, özellikle pigment epitel türevi faktör (PEDF), insülin benzeri büyüme faktörleri (IGF'ler) ve granülosit makrofaj-koloni uyarıcı faktör (GM-CSF) RPE hücrelerinin işlev bozukluğuna ve kaybına yol yorar, ardından fotoreceptör ve retinal ganglion hücre (RGC) ölümü3,4,5 . AMD metabolik, fonksiyonel, genetik ve çevresel faktörler arasındaki etkileşimden kaynaklanan karmaşık bir hastalıktır4. AAMD için tedavi eksikliği, sanayileşmiş ülkelerde 60 yaşından büyük hastalarda körlüğün başlıca nedenidir9,10. PEDF ve GM-CSF'yi aşırı ifade eden genetiği değiştirilmiş RPE hücrelerinin subretinal nakli ile nöroprotektif ve nörojenik retina ortamının yeniden birleşmesi, oksidatif stresin etkilerini azaltarak, iltihabı inhibe ederek ve hücre sağkalımını destekleyerek retina dejenerasyonunu önleme potansiyeline sahiptir11,12,13,14,15,16 . Genleri hücrelere ulaştırmak için çeşitli metodolojiler olsa da, pedf ve GM-CSF genlerini RPE hücrelerine ulaştırmak için viral olmayan hiperaktif Uyuyan Güzel transposon sistemini seçtik, çünkü güvenlik profili, genlerin konak hücrelerin genomuna entegrasyonu ve teslim edilen genleri transkripsiyonel olmayan aktif bölgelere entegre etme eğilimi nedeniyle17. 18,19.
Hücresel oksidatif stres, hidrojen peroksit (H 2O2),4-hidroinonenal (HNE), tertbutilhidroksit (tBH), yüksek oksijen gerginlikleri ve görünür ışık (tam spektrum veya UV ışınlama)20,21dahil olmak üzere çeşitli oksidatif ajanlar tarafından in vitro kültürlenen hücrelerde indüklenebilir. Yüksek oksijen gerilimleri ve ışık, diğer sistemlere aktarilebilirliği sınırlayan özel ekipman ve koşullar gerektirir. H 2 O2,HNE ve tBH gibi ajanlar üst üste binen oksidatif stres moleküler ve hücresel değişikliklere neden olarak. PEDF ve GM-CSF'nin antioksidan aktivitesini test etmek içinH2 O2'yi seçtik, çünkü RPE hücreleri tarafından fotoreceptör dış segment fagositoz22 sırasında reaktif oksijen ara olarak üretildiği ve in vivo23oküler dokularda bulunduğu için uygun ve biyolojik olarak alakalıdır. Glutatyonun oksidasyonu gözdeki H 2 O2üretiminden kısmen sorumlu olabileceğinden, çalışmalarımızda H 2 O2 kaynaklı oksidatif stres ve 21,22hücrelerinin rejeneratifkapasitesi ile bağlantılı GSH / glutatyon seviyelerini analizettik. Glutatyon seviyelerinin analizi özellikle gözdeki anti-oksidatif koruyucu mekanizmalara katıldığı içinönemlidir 24. H 2 O2'yemaruz kalma, RPE hücrelerinin 1 , 25 ,26,27 , 28, 29,30oksidatifstres duyarlılığını ve antioksidan aktivitesini incelemek için bir model olarak sıklıkla kullanılır ve ayrıca, "fizyolojik" bir oksidatif stres kaynağı olan ışık kaynaklı oksidatif stres hasarı ile benzerlikler gösterir21.
Nöroprotektif faktörlerin işlevselliğini ve etkinliğini değerlendirmek için, analizin PEDF ve GM-CSF'yi aşırı ifade etmek için genetiği değiştirilmiş hücreler tarafından ifade edilen büyüme faktörlerinin anti-oksidatif etkisini ölçmesini sağlayan bir in vitro model geliştirdik. Burada, PEDF ve GM-CSF genleri ile transfected RPE hücrelerinin H 2 O2'ninzararlı etkilerine transfected olmayan kontrol hücrelerinden daha dirençli olduğunu gösteriyoruz, hücre morfolojisi, yoğunluğu, canlılığı, hücre içi glutatyon seviyesi ve reaktif oksijen türlerini azalttığı gösterilen mitokondriyal ayrıştırma proteini 2'yi kodlayan UCP2 geninin ekspresyonu ile kanıtlandı (ROS)31.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>24-well plates</td>
			<td>Corning</td>
			<td>353047</td>
			<td></td>
		</tr>
		<tr>
			<td>6-well plates</td>
			<td>Greiner</td>
			<td>7657160</td>
			<td></td>
		</tr>
		<tr>
			<td>96-well culture plate white with clear flat bottom&#160;</td>
			<td>Costar</td>
			<td>3610</td>
			<td>Allows to check the cells before measuring the luminescence (GSH-Glo Assay)</td>
		</tr>
		<tr>
			<td>96-well plates</td>
			<td>Corning&#160;</td>
			<td>353072</td>
			<td></td>
		</tr>
		<tr>
			<td>Acrylamid 40%</td>
			<td>Biorad</td>
			<td>161-0144</td>
			<td></td>
		</tr>
		<tr>
			<td>Amphotericin B</td>
			<td>AMIMED</td>
			<td>4-05F00-H</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-GMCSF</td>
			<td>ThermoFisher Scientific</td>
			<td>PA5-24184</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-mouse IgG/IgA/IgM&#160;</td>
			<td>Agilent</td>
			<td>P0260</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-PEDF&#160;</td>
			<td>Santa Cruz Biotechnology Inc</td>
			<td>sc-390172</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-penta-His&#160;</td>
			<td>Qiagen</td>
			<td>34660</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-phospho-Akt</td>
			<td>Cell Signaling Technology</td>
			<td>9271</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-rabbit IgG H&#38;L-HRP</td>
			<td>Abcam</td>
			<td>ab6721</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody donkey anti-rabbit Alexa Fluor 594&#160;</td>
			<td>ThermoFisher Scientific&#160;</td>
			<td>A11034</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody goat anti-mouse Alexa 488</td>
			<td>ThermoFisher Scientific</td>
			<td>A-11029</td>
			<td></td>
		</tr>
		<tr>
			<td>ARPE-19 cell line</td>
			<td>ATCC</td>
			<td>CRL-2302</td>
			<td></td>
		</tr>
		<tr>
			<td>BSA&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>A9418-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>chamber culture glass slides</td>
			<td>Corning</td>
			<td>354118</td>
			<td></td>
		</tr>
		<tr>
			<td>CytoTox-Glo Cytotoxicity Assay&#160;</td>
			<td>Promega</td>
			<td>G9291</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Sigma-Aldrich</td>
			<td>D9542-5MG</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/Ham`s F12&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>D8062</td>
			<td></td>
		</tr>
		<tr>
			<td>Duo Set ELISA kit</td>
			<td>R&#38;D Systems&#160;</td>
			<td>DY215-05</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>ThermoFisher Scientific</td>
			<td>78440</td>
			<td></td>
		</tr>
		<tr>
			<td>ELISAquant kit</td>
			<td>BioProducts MD</td>
			<td>PED613-10-Human</td>
			<td></td>
		</tr>
		<tr>
			<td>Eyes (human)</td>
			<td>Lions Gift of Sight Eye Bank (Saint Paul, MN)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>FBS&#160;</td>
			<td>Brunschwig</td>
			<td>P40-37500</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluoromount Aqueous Mounting Medium</td>
			<td>Sigma-Aldrich</td>
			<td>F4680-25ML</td>
			<td></td>
		</tr>
		<tr>
			<td>FLUOstar Omega plate reader&#160;</td>
			<td>BMG Labtech</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>GraphPad Prism software (version 8.0)</td>
			<td>GraphPad Software, Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>GSH-Glo Glutathione Assay</td>
			<td>Promega</td>
			<td>V6912</td>
			<td></td>
		</tr>
		<tr>
			<td>hydrogen peroxide (H2O2)</td>
			<td>Merck</td>
			<td>107209</td>
			<td></td>
		</tr>
		<tr>
			<td>ImageJ software (image processing program)</td>
			<td>W.S. Rasband, NIH, Bethesda, MD, USA; https://imagej.nih.gov/ij/; 1997&#8211;2014</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Imidazol&#160;</td>
			<td>Axonlab</td>
			<td>A1378.0010</td>
			<td></td>
		</tr>
		<tr>
			<td>Leica DMI4000B microscope&#160;</td>
			<td>Leica Microsystems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LightCycler 480 Instrument II&#160;</td>
			<td>Roche Molecular Systems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LightCycler 480 SW1.5.1 software</td>
			<td>Roche Molecular Systems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma-Aldrich</td>
			<td>71376-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>NaH2PO4</td>
			<td>Axonlab</td>
			<td>3468.1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Neon Transfection System&#160;</td>
			<td>ThermoFisher Scientific</td>
			<td>MPK5000</td>
			<td></td>
		</tr>
		<tr>
			<td>Neon Transfection System 10 &#181;L Kit</td>
			<td>ThermoFisher Scientific</td>
			<td>MPK1096</td>
			<td></td>
		</tr>
		<tr>
			<td>Neubauer chamber</td>
			<td>Marienfeld-superior</td>
			<td>640010</td>
			<td></td>
		</tr>
		<tr>
			<td>Ni-NTA superflow&#160;</td>
			<td>Qiagen</td>
			<td>30410</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrocellulose&#160;</td>
			<td>VWR</td>
			<td>732-3197</td>
			<td></td>
		</tr>
		<tr>
			<td>Omega Lum G Gel Imaging System</td>
			<td>Aplegen Life Science</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PBS 1X</td>
			<td>Sigma-Aldrich</td>
			<td>D8537</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin</td>
			<td>Sigma-Aldrich</td>
			<td>P0781-100</td>
			<td></td>
		</tr>
		<tr>
			<td>PerfeCTa SYBR Green FastMix</td>
			<td>Quantabio</td>
			<td>95072-012</td>
			<td></td>
		</tr>
		<tr>
			<td>PFA&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>158127-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Pierce BCA Protein Assay Kit&#160;</td>
			<td>ThermoFisher Scientific</td>
			<td>23227</td>
			<td></td>
		</tr>
		<tr>
			<td>Primers&#160;</td>
			<td>Invitrogen&#160;</td>
			<td></td>
			<td>See Table 1 in Supplementary Materials</td>
		</tr>
		<tr>
			<td>pSB100X (250 ng/&#181;L)</td>
			<td></td>
			<td></td>
			<td>M&#225;t&#233;s et al., 2009. Provide by Prof. Zsuzsanna Izsvak</td>
		</tr>
		<tr>
			<td>pT2-CMV-GMCSF-His plasmid DNA (250 ng/&#181;L)</td>
			<td></td>
			<td></td>
			<td>Constructed using the existing pT2-CMV-PEDF-EGFP plasmid reported in Johnen, S. et al. (2012) IOVS, 53 (8), 4787-4796.</td>
		</tr>
		<tr>
			<td>pT2-CMV-PEDF-His plasmid DNA (250 ng/&#181;L)</td>
			<td></td>
			<td></td>
			<td>Constructed using the existing pT2-CMV-PEDF-EGFP plasmid reported in Johnen, S. et al. (2012) IOVS, 53 (8), 4787-4796.</td>
		</tr>
		<tr>
			<td>QIAamp DNA Mini Kit</td>
			<td>QIAGEN</td>
			<td>51304</td>
			<td></td>
		</tr>
		<tr>
			<td>recombinant hGM-CSF&#160;</td>
			<td>Peprotech</td>
			<td>100-11</td>
			<td></td>
		</tr>
		<tr>
			<td>recombinant hPEDF&#160;&#160;</td>
			<td>BioProductsMD</td>
			<td>004-096</td>
			<td></td>
		</tr>
		<tr>
			<td>ReliaPrep RNA Cell Miniprep System</td>
			<td>Promega</td>
			<td>Z6011</td>
			<td></td>
		</tr>
		<tr>
			<td>RIPA buffer</td>
			<td>ThermoFisher Scientific</td>
			<td>89901</td>
			<td></td>
		</tr>
		<tr>
			<td>RNase-free DNase Set</td>
			<td>QIAGEN</td>
			<td>79254</td>
			<td></td>
		</tr>
		<tr>
			<td>RNeasy Mini Kit</td>
			<td>QIAGEN</td>
			<td>74204</td>
			<td></td>
		</tr>
		<tr>
			<td>SDS</td>
			<td>Applichem</td>
			<td>A2572</td>
			<td></td>
		</tr>
		<tr>
			<td>Semi-dry transfer system for WB&#160;</td>
			<td>Bio-Rad</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>SuperMix qScript</td>
			<td>Quantabio</td>
			<td>95048-025</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-buffered saline (TBS)&#160;</td>
			<td>ThermoFisher Scientific</td>
			<td>15504020</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>AppliChem</td>
			<td>A4975</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin/EDTA</td>
			<td>Sigma-Aldrich</td>
			<td>T4174</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween</td>
			<td>AppliChem&#160;</td>
			<td>A1390</td>
			<td></td>
		</tr>
		<tr>
			<td>Urea</td>
			<td>ThermoFisher Scientific</td>
			<td>29700</td>
			<td></td>
		</tr>
		<tr>
			<td>WesternBright ECL HRP substrate</td>
			<td>Advansta</td>
			<td>K-12045-D50</td>
			<td></td>
		</tr>
		<tr>
			<td>Whatman nitrocellulose membrane</td>
			<td>Chemie Brunschwig</td>
			<td>MNSC04530301</td>
			<td></td>
		</tr>
	</tbody>
</table>

induction and analysis of oxidative stress in sleeping beauty transposon-transfected human retinal pigment epithelial cells

Oksidatif stres, dünya çapında ~30 milyon hastayı etkileyen bir patoloji olan yaşa bağlı makula dejenerasyonu (AMD) da dahil olmak üzere çeşitli dejeneratif hastalıklarda kritik bir rol oynamaktadır. Retina pigment epitelinde (RPE)- sentezlenmiş nöroprotektif faktörlerde, örneğin pigment epitel türevi faktör (PEDF) ve granülosit-makrofaj koloni uyarıcı faktöründe (GM-CSF) azalmaya, ardından RPE hücrelerinin kaybına ve sonunda fotoreceptör ve retina ganglion hücre (RGC) ölümüne yol açar. PeDF ve GM-CSF'yi aşırı ifade eden transkreinal RPE hücrelerinin subretinal nakli ile nöroprotektif ve nörojenik retina ortamının yeniden kavrunun, oksidatif stresin etkilerini azaltarak, iltihabı inhibe ederek ve hücre sağkalımını destekleyerek retina dejenerasyonunu önleme potansiyeline sahip olduğunu hipotez ediyoruz. Uyuyan Güzel transposon sistemi (SB100X) insan RPE hücreleri PEDF ve GM-CSF genleri ile transfekte edilmiştir ve qPCR, batı lekesi, ELISA ve immünofluoresans kullanılarak kararlı gen entegrasyonu, uzun süreli gen ekspresyonu ve protein salgılanması gösterilmiştir. Transfected RPE hücreleri tarafından salgılanan PEDF ve GM-CSF'nin işlevselliğini ve gücünü doğrulamak için, kültürdeki RPE hücreleri üzerinde H2O2kaynaklı oksidatif stresin azaltılmasını ölçmek için bir in vitro test geliştirdik. Hücre koruması hücre morfolojisi, yoğunluğu, hücre içi glutatyon düzeyi, UCP2 gen ekspresyonu ve hücre canlılığı analiz ederek değerlendirildi. Her ikisi de, PEDF ve/veya GM-CSF'yi aşırı ifade eden transfected RPE hücreleri ve transknakte olmayan ancak PEDF ve/veya GM-CSF ile önceden tedavi edilen hücreler (ticari olarak mevcut veya transfected hücrelerden arındırılmış) tedavi edilmeyen kontrollere kıyasla önemli antioksidan hücre koruması göstermiştir. Mevcut H2O2-modeli, AMD veya benzeri nörodejeneratif hastalıkları tedavi etmek için etkili olabilecek faktörlerin antioksidan etkisini değerlendirmek için basit ve etkili bir yaklaşımdır.

İnsan Retina Pigment Epitel hücrelerinde oksidatif stresin indüksiyonu ARPE-19 ve primer hRPE hücreleri24saat boyunca değişen H 2 O2 konsantrasyonları ile tedavi edildi ve antioksidan glutatyonun hücre içi seviyesi ölçüldü (Şekil 2A,B). H2O2 50 μM ve 100 μM glutatyon üretimini etkilemezken, 350 μM'de ARPE-19 ve primer hRPE hücrelerinde glutatyonda önemli bir a...

Watch this Scientific Journal Video about Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells at JoVE.com

Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells

Hücre morfolojisi, canlılık, yoğunluk, glutatyon ve UCP-2 seviyesini analiz eden retina pigment epitel hücrelerini H2O2ile tedavi ederek oksidatif stres modelinin geliştirilmesi ve kullanımı için bir protokol sunuyoruz. Transposon transfected hücreler tarafından salgılanan proteinlerin nöroretinal dejenerasyonu tedavi etmek için antioksidan etkisini araştırmak için yararlı bir modeldir.

Uyuyan Güzel Transposon Transposon Transfected İnsan Retina Pigment Epitel Hücrelerinde Oksidatif Stresin İndüksiyonu ve Analizi

Oxidative stress plays a critical role in several degenerative diseases, including age-related macular degeneration (AMD), a pathology that affects ~30 ...

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Medicine

Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells

Identification of Sleeping Beauty Transposon Insertions in Solid Tumors using Linker-mediated PCR

Identification of Sleeping Beauty Transposon Insertions in Solid Tumors using Linker-mediated PCR

Tanımlanması<em&gt; Sleeping Beauty</emLinker-aracılı PCR kullanarak Solid Tümörlerde&gt; Transposon Eklemeler

Genomic, proteomic, transcriptomic, and epigenomic analyses of human tumors indicate that there are thousands of anomalies within each cancer genome ...

Transcriptomic Analysis of Human Retinal Surgical Specimens Using jouRNAl

Journal kullanarak İnsan Retina Cerrahi Örneklerinin transkriptomik Analizi

Retinal detachment (RD) describes a separation of the neurosensory retina  from the retinal pigmented epithelium (RPE). The RPE is essential for normal  ...

Isolation, Cryopreservation and Culture of Human Amnion Epithelial Cells for Clinical Applications

İzolasyon, Kriyoprezervasyon ve Kültür Klinik Uygulamaları İnsan Amniyon Epitel Hücreleri

Human amnion epithelial cells (hAECs) derived from term or pre-term amnion membranes have attracted attention from researchers and clinicians as a ...

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

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 ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pankreas Kanseri Yeni Tedavi Stratejileri Geliştirme Hücre Özellikleri Kök incelenmesi

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

A Step by Step Protocol for Subretinal Surgery in Rabbits

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 ...

Depletion of Mouse Cells from Human Tumor Xenografts Significantly Improves Downstream Analysis of Target Cells

İnsan Tümör ksenogratflarının gelen Fare Hücrelerinin tükenmesi Anlamlı Hedef Hücre Alt Analizi geliştirir

The use of in vitro cell line models for cancer research has been a useful tool. However, it has been shown that these models fail to reliably mimic ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Farelerde Hızlandırılmış Ateroskleroz Indüksiyonu: "Tel-Yaralanma" Modeli

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Fosfor-31 Manyetik Rezonans Spektroskopisi: Ölçme için Aracı<em&gt; İn Vivo</emİnsan İskelet Kası içinde&gt; Mitokondriyal Oksidatif Fosforilasyon Kapasitesi

Skeletal muscle mitochondrial oxidative phosphorylation (OXPHOS) capacity, which is critically important in health and disease, can be measured in vivo ...