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

저자는 훌륭한 기술 지원에 대한 그레그 실리와 알레인 콘티와 베를린의 맥스 델브뤼크 센터에서 교수 Zsuzsanna 이즈바크에게 감사하고 싶습니다 pSB100X 및 pT2-CAGGS-비너스 플라스미드를 친절하게 제공합니다. 이 작품은 스위스 국립 과학 재단과 유럽 위원회가 제7 프레임 워크 프로그램의 맥락에서 지원되었습니다. Z.I는 유럽 연구 위원회, ERC 고급 [ERC-2011-ADG 294742]에 의해 투자되었다.

인간의 눈의 수집 및 사용에 대한 절차는 연구를위한 주 윤리위원회에 의해 승인되었다 (아니. 2016-01726).
1. 세포 격리 및 배양 조건
<ol><li>인간 ARPE-19 세포주<ol>
<li>문화 5 x 105 ARPE-19 세포, 인간 RPE 세포주, 덜벡코의 수정 된 독수리의 중간 / 영양 혼합물 F-12 햄 (DMEM / 햄의 F-12)은 10 % 태아 소 혈청 (FBS), 80 U / mL 페니실린, 80 U / mL 페니실린으로 보충 80 μg/mL 연쇄상 구균, 및...

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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=Modulation+of+oxidative+stress+as+an+anticancer+strategy.">Modulation of oxidative stress as an anticancer strategy.</a> Nature Reviews Drug Discovery. 12, 931-947 (2013).</li><li>Wang, X., 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=PEDF+protects+human+retinal+pigment+epithelial+cells+against+oxidative+stress+via+upregulation+of+UCP2+expression.">PEDF protects human retinal pigment epithelial cells against oxidative stress via upregulation of UCP2 expression.</a> Molecular Medicine Reports. 19 (1), 59-74 (2019).</li><li>Donadelli, M., Dando, I., Fiorini, C., Palmieri, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=UCP2,+a+mitochondrial+protein+regulated+at+multiple+levels.">UCP2, a mitochondrial protein regulated at multiple levels.</a> Cellular and Molecular Life Sciences. 71, 1171-1190 (2014).</li></ol>

여기에 제시된 프로토콜은 어떤 putative 유익한 유전자로 전염되는 세포에 적용될 수 있는 전감염된 세포에 의해 생성된 PEDF 및 GM-CSF의 산화 및 보호 기능을 분석하는 접근법을 제안합니다. 유전자 변형 세포를 이식하여 조직에 단백질을 전달하는 것을 목표로 하는 유전자 치료 전략에서, 단백질 발현의 수준, 발현의 장수 및 질병의 모델에서 발현 된 단백질의 효과에 대한 정보를 얻는 것이 중요하...

여기에 설명된 모델은 세포의 산화 스트레스를 줄이기 위한 바이오 의약품 제제의 효율성을 평가하는 유용한 접근법을 제공합니다. 우리는 H2O2-매개산화 용 안료 상피 세포에 대한 PEDF 및 GM-CSF의 보호 효과를 조사하기 위해 모델을 사용했으며, 이는 높은 수준의 O2에노출되며, 가시광선, 광수용체 외부 세그먼트 멤브레인의 phagocytosis를 통해 반응성 산소 종(ROS)1, 1, 1, 1 2. 그(것)들은 혈관 나이 관련 황반 변성의 병인에 중요한 기여자로 간주됩니다 (aAMD)3,4,5,6,7,8. 게다가, RPE 합성 신경 보호 인자, 특히 안료 상피 유래 인자 (PEDF), 인슐린 과같은 성장 인자 (IGFs), 및 과립구 대식세포 -콜로니 자극 인자 (GM-CSF)가 RPE 세포의 기능 장애 및 손실로 이어지는후, 광수용체 및 망막 갱셀(R) 세포3,4GC세포(RGC) . AMD는 대사, 기능성, 유전적 및 환경적 요인4사이의 상호 작용에서 발생하는 복잡한 질환이다. aAMD에 대한 치료의 부족은 선진국9,10에서60 세 이상의 환자에서 실명의 주요 원인입니다. PEDF및 GM-CSF를 과도하게 발현하는 유전자 변형 RPE 세포의 피하 이식에 의한 신경보호 및 신경성 망막 환경의 재구성은 산화 스트레스의 효과를 완화하고 염증을 억제하고 세포생존11,12,13, 14,14, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15,15,1 . 세포에 유전자를 전달하는 몇 가지 방법론이 있지만, 우리는 그 안전 프로필, 숙주 세포의 게놈에 유전자의 통합, 그리고 우리가이전에보여 준 바와 같이 비 전사 활성 사이트에 전달 된 유전자를 통합하는 성향 때문에 RPE 세포에 PEDF 및 GM-CSF 유전자를 제공하기 위해 비 바이러스 성 활동적인 수면 뷰티 트랜스포슨 시스템을선택했습니다. 18,19.
세포 산화 스트레스는 과산화수소(H2O2),4-하이드로이네날(HNE), 테르부틸하이드로산화물(tBH), 높은 산소 장력, 가시광선(전체 스펙트럼 또는 UV 조사)20,21을포함한 여러 산화제에 의해 시험관내에서 배양되는 세포에서 유도될 수 있다. 높은 산소 장력과 빛은 다른 시스템으로의 전송 가능성을 제한하는 특수 장비 와 조건이 필요합니다. H2O2,HNE 및 tBH와 같은 제제는 겹치는 산화 스트레스 분자 및 세포 변화를 유도한다. 우리는 포토수용체 외부 세그먼트 phagocytosis(22)에서 반응성 산소 중간체로서 RPE 세포에 의해 생성되기 때문에 편리하고 생물학적으로 관련성이 있기 때문에 PEDF 및 GM-CSF의 항산화 활성을테스트하기 위해 H2O2를 선택했으며 생체내안구 조직에서 발견된다. 글루타티온의 산화는 눈에 H2O2의 생산에 부분적으로 책임이 있을 수 있기 때문에, 우리는 H2 O2-유도산화스트레스및세포의 재생 능력에 연결되는 우리의 연구에서 GSH/글루타티온의 수준을 분석하였다21,22. 글루타티온 수준의 분석은 눈24의산화 방지 메커니즘에 참여하기 때문에 특히 관련이 있다. H2O2에노출되는 것은 RPE 세포의 산화 스트레스 감수성 및 항산화활성을검사하기 위한 모델로 자주 사용되며, 또한, 광유발 산화 스트레스 손상, 산화 스트레스21의"생리적" 원인과 유사성을 나타낸다. 
신경 보호 요인의 기능성과 효과를 평가하기 위해, 우리는 분석이 PEDF와 GM-CSF를 과발현하기 위하여 유전자 변형된 세포에 의해 표현된 성장 인자의 산화 효과를 정량화하는 분석을 허용하는 시험관 내 모형을 개발했습니다. 여기서, 우리는 PEDF및 GM-CSF에 대한 유전자와 함께 전염된 RPE 세포가 세포 형태, 밀도, 생존성, 글루타티온의 세포내 수준 및 UCP2 유전자의 발현에 의해 입증된 바와 같이, 비과감염 대조구 세포보다 H2O2의 유해한 영향에 더 저항력이 있음을 보여주며, 이는 미토콘드리아 분리 단백질(ROS31)에 대한 코드로 인해31종의산소를 감소시키는 것으로 나타났다.

<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

산화 스트레스는 전 세계적으로 3천만 명의 환자에게 영향을 미치는 병리학인 노화 관련 황반 변성(AMD)을 포함한 여러 퇴행성 질환에서 중요한 역할을 합니다. 망막 색소 상피(RPE)-합성 신경보호인자, 예를 들어, 안료 상피 유래 인자(PEDF) 및 과립세포-대식세포 콜로니-자극인자(GM-CSF)의 감소로 이어지며, 결국 광수용체 및 망막 갱셀(RGC)의 손실로 이어진다. 우리는 PEDF와 GM-CSF를 과발현하는 과발뇌형 RPE 세포의 감속 이식에 의한 신경보호 및 신경성 망막 환경의 재구성이 산화 스트레스의 효과를 완화하고 염증을 억제하며 세포 생존을 지원함으로써 망막 변성을 예방할 수 있는 잠재력을 가지고 있다고 가설합니다. 수면미트랜스포손 시스템(SB100X)을이용하여 인간 RPE 세포는 PEDF 및 GM-CSF 유전자에 감염되어 qPCR, 서부 블롯, ELISA 및 면역불률을 이용한 안정적인 유전자 통합, 장기 유전자 발현 및 단백질 분비를 보였다. 전감염된 RPE 세포에 의해 분비된 PEDF 및 GM-CSF의 기능성 및 효능을 확인하기 위해, 배양시 RPE 세포에 대한H2O2-유도산화 스트레스의 감소를 정량화하기 위한 시험관 내 분석서를 개발하였다. 세포 보호는 세포 형태학, 밀도, 글루타티온의 세포 내 수준, UCP2 유전자 발현 및 세포 생존가능성을 분석하여 평가하였다. 둘 다, PEDF 및/또는 GM-CSF 및 세포를 과도하게 발현하는 전감염된 RPE 세포는 비감염되었지만 PEDF 및/또는 GM-CSF(소판 또는 전감염된 세포로부터 정수)로 전처리된 비처리 대조군과 비교하여 상당한 항산화 세포 보호를 보였다. 본H2O2-모델은AMD 또는 이와 유사한 신경퇴행성 질환을 치료하는 데 효과적일 수 있는 요인의 항산화 효과를 평가하는 간단하고 효과적인 접근법이다.

인간 망막 안료 상피 세포에서 산화 스트레스유도 ARPE-19 및 1차 hRPE 세포는 24시간 동안H2 O2의다양한 농도로 처리되었고 항산화 글루타티온의 세포내 수준을 정량화하였다(도2A,B). H2O2 앳 50 μM 및 100 μM은 글루타티온 생산에 영향을 미치지 않았지만 350 μM에서는 ARPE-19 및 1차 hRPE 세포에서 글?...

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

우리는 H 2O2로망막 색소 상피 세포를 치료하여 세포 형태, 생존가능성, 밀도, 글루타티온 및 UCP-2 수준의 개발 및 사용을 위한 프로토콜을 제시합니다. 신경망 변성을 치료하기 위해 트랜스포슨-감염된 세포에 의해 분비되는 단백질의 항산화 효과를 조사하는 데 유용한 모델이다.

수면 의 아름다움 트랜스포슨 - 전염 인간 망막 안료 상피 세포에서 산화 스트레스의 유도 및 분석

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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JoVE Journal

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

의 식별<em&gt; 잠자는 숲속의 미녀</em링커로 인한 PCR을 사용하여 고체 종양의&gt; Transposon의 삽입

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

연구

의학

Transcriptomic Analysis of Human Retinal Surgical Specimens Using jouRNAl

저널을 사용하여 인간의 망막 수술 표본의 Transcriptomic 분석

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

분리, 냉동 보존 및 문화 임상 응용 프로그램에 대한 인간 양막 상피 세포

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

서스펜션에 망막 색소 상피 세포를 제공하기 위해 설치류에서 망막 주사를 수행

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

췌장암 새로운 치료 전략을 개발하기위한 전지 특성 줄기 공부

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

토끼의 망막 수술에 대한 단계 프로토콜에 의해 단계

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

인간 종양 이종 이식에서 마우스 세포의 고갈은 크게 대상 세포의 다운 스트림 분석을 향상

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

마우스에서 가속 죽상 경화증의 유도: "와이어 부상" 모델

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

인-31 자기 공명 분광학 : 측정을위한 도구<em&gt; 생체</em인간의 골격 근육에&gt; 미토콘드리아 산화 적 인산화 능력

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