Name: microrna expression profiles of human ips cells, retinal pigment epithelium derived from ips, and fetal retinal pigment epithelium
Uploaded: 2014-06-24T13:00:00
Description: Watch this Scientific Journal Video about MicroRNA Expression Profiles of Human iPS Cells, Retinal Pigment Epithelium Derived From iPS, and Fetal Retinal Pigment Epithelium at JoVE.com

此处包含的意见或主张是作者的个人观点，并不应被理解为官方或反映了陆军部和国防部的意见。 这项研究进行的同时，作者惠特尼A.格林，阿尔贝托穆尼斯和拉梅什R.凯妮召开全国研究理事会博士后研究员院士在USAISR。 微阵列分析是由Greehey儿童癌症研究所基因芯片核心设备和生物信息学系UT健康科学中心圣安东尼奥进行。 这项工作是由美国陆军临床康复医学研究发展计划（CRMRP）和军事作战医学研究发展计划（MOMRP）的支持。

培养试剂和培养板的1。准备<ol><li> mTeSR1媒体：根据制造商的指示准备mTeSR1媒体。解冻百毫升mTeSR1 5倍的补充一夜之间在4°C。添加百毫升mTeSR1 5倍的补充至400ml mTeSR1基础培养基中拌匀。此介质是稳定的，在4℃下进行长达2周，并在-20℃下6个月。 </li><li>分化培养基：准备分化培养基中的DMEM/F12，得到0.1mM的β-巯基乙醇，0.1mM非必需氨基酸，2.0 2mM L-谷氨酰胺，10％敲除血清，和10微克/毫升庆大霉素。 </l...

<ol>
	<li>Yu, Z., 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=miRNAs+regulate+stem+cell+self-renewal+and+differentiation.">miRNAs regulate stem cell self-renewal and differentiation.</a> Front Genet. 3, 191 (2012).</li><li>Romano, G., 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=A+commentary+on+iPS+cell+biology:+Potential+applications+in+autologous+tissue+and+cell+transplantation,+development+of+new+models+for+the+study+of+human+pathological+conditions+and+drug+screening.">A commentary on iPS cell biology: Potential applications in autologous tissue and cell transplantation, development of new models for the study of human pathological conditions and drug screening.</a> J Cell Physiol. 229, 148-152 (2014).</li><li>Almeida, P. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunogenicity+of+pluripotent+stem+cells+and+their+derivatives.">Immunogenicity of pluripotent stem cells and their derivatives.</a> Circ Res. 112, 549-561 (2013).</li><li>Buchholz, D. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Derivation+of+functional+retinal+pigmented+epithelium+from+induced+pluripotent+stem+cells.">Derivation of functional retinal pigmented epithelium from induced pluripotent stem cells.</a> Stem Cells. 27, 2427-2434 (2009).</li><li>Kokkinaki, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+induced+pluripotent+stem-derived+retinal+pigment+epithelium+(RPE)+cells+exhibit+ion+transport,+membrane+potential,+polarized+vascular+endothelial+growth+factor+secretion,+and+gene+expression+pattern+similar+to+native+RPE.">Human induced pluripotent stem-derived retinal pigment epithelium (RPE) cells exhibit ion transport, membrane potential, polarized vascular endothelial growth factor secretion, and gene expression pattern similar to native RPE.</a> Stem Cells. 29, 825-835 (2011).</li><li>Bharti, K., 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=The+new+paradigm:+retinal+pigment+epithelium+cells+generated+from+embryonic+or+induced+pluripotent+stem+cells.">The new paradigm: retinal pigment epithelium cells generated from embryonic or induced pluripotent stem cells.</a> Pigment Cell Melanoma Res. 24, 21-34 (2011).</li><li>Park, I. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reprogramming+of+human+somatic+cells+to+pluripotency+with+defined+factors.">Reprogramming of human somatic cells to pluripotency with defined factors.</a> Nature. 451, 141-146 (2008).</li><li>Pera, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stem+cells.+A+new+year+and+a+new+era.">Stem cells. A new year and a new era.</a> Nature. 451, 135-136 (2008).</li><li>Subba Rao, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thinking+outside+the+liver:+Induced+pluripotent+stem+cells+for+hepatic+applications.">Thinking outside the liver: Induced pluripotent stem cells for hepatic applications.</a> World J Gastroenterol. 19, 3385-3396 (2013).</li><li>Yoshida, Y., Yamanaka, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=iPS+cells:+a+source+of+cardiac+regeneration.">iPS cells: a source of cardiac regeneration.</a> J Mol Cell Cardiol. 50, 327-332 (2011).</li><li>Lee, K. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+sensory+neurons+derived+from+induced+pluripotent+stem+cells+support+varicella-zoster+virus+infection.">Human sensory neurons derived from induced pluripotent stem cells support varicella-zoster virus infection.</a> PLoS One. 7, (2012).</li><li>Alipio, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversal+of+hyperglycemia+in+diabetic+mouse+models+using+induced-pluripotent+stem+(iPS)-derived+pancreatic+beta-like+cells.">Reversal of hyperglycemia in diabetic mouse models using induced-pluripotent stem (iPS)-derived pancreatic beta-like cells.</a> Proc Natl Acad Sci U S A. 107, 13426-13431 (2010).</li><li>la Cour, M., Tezel, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+retinal+pigment+epithelium.">The retinal pigment epithelium.</a> Adv Organ Biol. 10, 253-272 (2005).</li><li>Carr, A. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+effects+of+human+iPS-derived+retinal+pigment+epithelium+cell+transplantation+in+the+retinal+dystrophic+rat.">Protective effects of human iPS-derived retinal pigment epithelium cell transplantation in the retinal dystrophic rat.</a> PLoS One. 4, (2009).</li><li>Schwartz, S. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Embryonic+stem+cell+trials+for+macular+degeneration:+a+preliminary+report.">Embryonic stem cell trials for macular degeneration: a preliminary report.</a> Lancet. 379, 713-720 (2012).</li><li>Muniz, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinoid+uptake,+processing+and+secretion+in+human+iPS-RPE+support+the+visual+cycle.">Retinoid uptake, processing and secretion in human iPS-RPE support the visual cycle.</a> Invest Ophthalmol Vis Sci. 55, 198-209 (2014).</li><li>Leonardo, T. R., 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=The+functions+of+microRNAs+in+pluripotency+and+reprogramming.">The functions of microRNAs in pluripotency and reprogramming.</a> Nat Cell Biol. 14, 1114-1121 (2012).</li><li>Arora, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prediction+of+microRNAs+affecting+mRNA+expression+during+retinal+development.">Prediction of microRNAs affecting mRNA expression during retinal development.</a> BMC Dev Biol. 10, (2010).</li><li>Huntzinger, E., Izaurralde, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+silencing+by+microRNAs:+contributions+of+translational+repression+and+mRNA+decay.">Gene silencing by microRNAs: contributions of translational repression and mRNA decay.</a> Nat Rev Genet. 12, 99-110 (2011).</li><li>Mallanna, S. K., Rizzino, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emerging+roles+of+microRNAs+in+the+control+of+embryonic+stem+cells+and+the+generation+of+induced+pluripotent+stem+cells.">Emerging roles of microRNAs in the control of embryonic stem cells and the generation of induced pluripotent stem cells.</a> Dev Biol. 344, 16-25 (2010).</li><li>Guo, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mammalian+microRNAs+predominantly+act+to+decrease+target+mRNA+levels.">Mammalian microRNAs predominantly act to decrease target mRNA levels.</a> Nature. 466, 835-840 (2010).</li><li>Adijanto, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microphthalmia-associated+transcription+factor+(MITF)+promotes+differentiation+of+human+retinal+pigment+epithelium+(RPE)+by+regulating+microRNAs-204/211+expression.">Microphthalmia-associated transcription factor (MITF) promotes differentiation of human retinal pigment epithelium (RPE) by regulating microRNAs-204/211 expression.</a> J Biol Chem. 287, 20491-20503 (2012).</li><li>Hu, G., 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=Identification+of+miRNA+signatures+during+the+differentiation+of+hESCs+into+retinal+pigment+epithelial+cells.">Identification of miRNA signatures during the differentiation of hESCs into retinal pigment epithelial cells.</a> PLoS One. 7, (2012).</li><li>Wilson, K. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MicroRNA+profiling+of+human-induced+pluripotent+stem+cells.">MicroRNA profiling of human-induced pluripotent stem cells.</a> Stem Cells Dev. 18, 749-758 (2009).</li><li>Neveu, P., 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=MicroRNA+profiling+reveals+two+distinct+p53-related+human+pluripotent+stem+cell+states.">MicroRNA profiling reveals two distinct p53-related human pluripotent stem cell states.</a> Cell Stem Cell. 7, 671-681 (2010).</li><li>Maminishkis, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Confluent+monolayers+of+cultured+human+fetal+retinal+pigment+epithelium+exhibit+morphology+and+physiology+of+native+tissue.">Confluent monolayers of cultured human fetal retinal pigment epithelium exhibit morphology and physiology of native tissue.</a> Invest Ophthalmol Vis Sci. 47, 3612-3624 (2006).</li><li>Qiagen, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=miRNeasy+Mini+Handbook.">miRNeasy Mini Handbook.</a> Sample and Assay Technologies. , (2013).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microarray+System+with+miRNA+Complete+Labeling+and+Hyb+Kit.">Microarray System with miRNA Complete Labeling and Hyb Kit.</a> Agilent Technologies. , (2011).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintenance+of+hESCs+and+hiPSCs+in+mTESR1+and+mTESR2.">Maintenance of hESCs and hiPSCs in mTESR1 and mTESR2.</a> Stem Cell Technologies. , (2010).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Analysis+of+DNA+or+RNA+using+its+wavelengths:+230+nm,+260+nm,+280+nm.">The Analysis of DNA or RNA using its wavelengths: 230 nm, 260 nm, 280 nm.</a> About Biotechnology. , (2013).</li></ol>

总之，这份报告描述了用于培养iPS细胞的方法，能干-RPE和胎儿的RPE。从视网膜色素上皮衍生的iPS在形态和功能上类似于胎儿视网膜色素上皮。上IPS-RPE也表达特性的RPE基因，包括RPE65，CRALBP，PEDF和LRAT 16。为了进一步表征这些细胞，提取RNA，并用于进行微阵列分析，以确定差异表达的miRNA。的信号强度的分析表明，在胎儿的RPE检测与的iPS-RPE比较的差异表达的miRNA的最大数目，暗示的iPS-RPE可?...

干细胞具有无限制和分化成任何体细胞类型的潜力来复制的能力。的技术，体细胞重编程为多能干细胞的发展，就引起了极大的热情在研究界和临床医生之间，作为个性化的组织再生的出现在地平线上1。诱导多能干细胞（iPS细胞）表现出无限的复制能力和多能性的相同的功能，胚胎干细胞（ES细胞），同时规避与胚胎干细胞相关的伦理困境。此外，病人的干细胞不会刺激免疫反应，大大增加了成功的治疗性应用2-3的概率。根据特定的培养条件下，iPS细胞已经显示出分化成体外几种不同的细胞类型，包括心肌细胞，神经细胞，胰腺β细胞，肝细胞和视网膜色素上皮（RPE）4-12。 在RPE是一个专门的层设在视网膜的后面，执行多种功能，这对于视力健康和功能，如杂散光，吞噬作用的光感受器外段的吸收必需的色素上皮细胞，和类视黄酸处理的生产视觉生色。视网膜色素上皮因损伤或疾病功能障碍深刻地影响感光健康和视觉功能就证明了致盲的疾病，是潜在的视网膜色素上皮病变，如年龄相关性黄斑变性（AMD），Stargardt病和色素性视网膜炎（RP）的结果13。可以恢复视力确实有效的治疗方法尚未实现，并更换视网膜色素上皮病变与正常RPE细胞可能是为了防止视力14-15损失的最佳选择。 RPE由iPS（IPS-RPE）是衍生的细胞的一个可能的来源，以取代受损视网膜色素上皮。的iPS-RPE表达characteris抽动RPE蛋白质LRAT，CRALBP，PEDF和RPE65;显示经典的高色素六角形的RPE形态;并执行RPE功能，如吞噬作用，视黄酸类化合物的处理，和11的分泌- 顺视网膜5,16。然而，之前的iPS-RPE可用于治疗中，IPS-RPE必须彻底特征。理解支配RPE分化的因素是必要的，以改善细胞用于临床应用的产率和纯度。 细胞分化是高度调节基因表达的结果。转录因子的基因组和协调的后生重构所需的分化和发育17期间发生的细胞命运的决定。信息核糖核酸（mRNA）的翻译者的microRNA（miRNA）的调控提出了调控的另一个层面，影响细胞命运1。 miRNA是短，约为22个核苷酸，核苷酸的长度，要么抑制翻译的结合到mRNA的3&#39;非编码区或靶mRNA的降解。的miRNA已经在几乎所有组织中被发现，而且迄今超过2,000人独特的microRNA已经注册在数据库中的miRBase。由于miRNA的要求只有部分互补结合到靶mRNA，单个的miRNA有可能绑定到几十或上百个目标，反之亦然， 即单个基因可以由几个不同的miRNA有针对性。这种混杂的结合特性大大提高了监管的复杂性，在细胞功能18-21各玩各的水平以及确定单个miRNA的功能的难易程度和作用。然而，研究表明，miRNA的通过影响DNA甲基化状态17细化分化过程中的基因表达。在一项研究更具体的RPE，miR-204/211被证明能促进视网膜色素上皮22的上皮细胞表型。另一组分析了miRNA的个人资料视网膜色素上皮的过程中由ES细胞分化，并透露组不同的miRNA在分化过程中23表示。事实上，miRNA表达谱可以明确区分的细胞类型，包括胚胎干细胞，前体细胞，和终末分化细胞24,25。超过250个miRNA的表达在视网膜上。基于这些研究，我们推测，RPE细胞或者由iPS分化过程中的miRNA中发挥重要作用。 本报告的目的是从IMR90-4的iPS描述为RPE的分化的协议细胞，收集的RNA的微阵列分析和微阵列数据的分析，以确定差异表达三种类型的细胞的iPS细胞的miRNA， ，的iPS-RPE细胞和胎儿的RPE。总RNA从每种细胞类型的培养物中提取和杂交的miRNA的微阵列，包含特异性针对1205人的miRNA和144病毒的miRNA探针。基因芯片结果进行比较d，来确定哪些miRNA的差异表达在不同的细胞类型。 miRNA的2倍或更大的表达倍数变化被选择用于进一步分析。甲miRNA的分析软件程序来鉴定差异表达的miRNA的潜在目标，并生成由所选择的miRNA调节的蜂窝网络。

<table border="0" cellpadding="0" cellspacing="0" width="676">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">mTeSR1 media + 5X supplement</td>
			<td style="width:156px;">Stem Cell Technologies</td>
			<td style="width:119px;">5850</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DMEM/F12</td>
			<td>Life Technologies</td>
			<td>11330-032</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-Mercaptoethanol</td>
			<td style="width:156px;">Sigma</td>
			<td style="width:119px;">M-7154</td>
			<td></td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:235px;">Non essential amino acids</td>
			<td style="width:156px;">Hyclone(Fisher)</td>
			<td>SH30853.01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Knockout serum replacement</td>
			<td style="width:156px;">Life Technologies</td>
			<td style="width:119px;">10828-028</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Gentamicin&#160;</td>
			<td style="width:156px;">Life Technologies</td>
			<td style="width:119px;">15750-060</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">MEM media</td>
			<td style="width:156px;">Life Technologies</td>
			<td style="width:119px;">10370-021</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">N1 supplement</td>
			<td style="width:156px;">Sigma</td>
			<td style="width:119px;">N-6530-5ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Taurine</td>
			<td style="width:156px;">Sigma</td>
			<td style="width:119px;">T-8691-25G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Hydrocortisone</td>
			<td style="width:156px;">Sigma</td>
			<td style="width:119px;">H0888-1G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Fetal bovine serum</td>
			<td style="width:156px;">Hyclone(Fisher)</td>
			<td>SH3008803HI</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triiodo-l-thyronine sodium salt</td>
			<td style="width:156px;">Sigma</td>
			<td style="width:119px;">T6397</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Sodium hydroxide</td>
			<td style="width:156px;">Sigma</td>
			<td style="width:119px;">S5881</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fetal RPE media RTEGM kit</td>
			<td style="width:156px;">Life Technologies</td>
			<td style="width:119px;">195406</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Dispase</td>
			<td style="width:156px;">Life Technologies</td>
			<td>17105-041</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Matrigel</td>
			<td style="width:156px;">BD Biosciences</td>
			<td style="width:119px;">354277</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Phosphate buffered saline</td>
			<td style="width:156px;">Hyclone(Fisher)</td>
			<td>10010-023</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Trypsin</td>
			<td style="width:156px;">Hyclone(Fisher)</td>
			<td style="width:119px;">25200-072</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Miltenyi Biotec washing buffer</td>
			<td style="width:156px;">StemGent</td>
			<td style="width:119px;">130-092-987</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Miltenyi Biotec rinsing buffer</td>
			<td style="width:156px;">StemGent</td>
			<td style="width:119px;">130-092-222</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Anti-TRA-1-60 microbead kit</td>
			<td style="width:156px;">StemGent</td>
			<td style="width:119px;">130-095-816</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Miltenyi Biotec cell sorter column</td>
			<td style="width:156px;">StemGent</td>
			<td style="width:119px;">130-021-101</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">RNeasy micro kit</td>
			<td style="width:156px;">Qiagen</td>
			<td style="width:119px;">74004</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">QIA shredder</td>
			<td style="width:156px;">Qiagen</td>
			<td style="width:119px;">79654</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">RNA 6000 Pico LabChip kit</td>
			<td style="width:156px;">Agilent</td>
			<td style="width:119px;">G2938-90046</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:235px;">Human miRNA microarray v16</td>
			<td style="width:156px;">Agilent</td>
			<td style="width:119px;">G4471A</td>
			<td></td>
		</tr>
	</tbody>
</table>

microrna expression profiles of human ips cells, retinal pigment epithelium derived from ips, and fetal retinal pigment epithelium

本报告的目的是描述的协议，用于比较的microRNA（miRNA）的人类诱导多能干细胞（iPS）细胞，视网膜色素上皮细胞（RPE）从人类iPS细胞（iPS-RPE）导出，胎儿视网膜色素上皮的配置文件。该协议包括收集的RNA通过微阵列分析，以及微阵列数据的分析，找出差异表达三种细胞类型的miRNA。该方法iPS细胞和胎儿视网膜色素上皮的文化解释。用于从人iPS分化的RPE的协议也被描述。我们描述了RNA提取技术被选中，允许非常小RNA在一个miRNA芯片的使用极大的恢复。最后，细胞通路和微阵列数据的网络分析解释。这些技术将促进三种不同类型的细胞的miRNA表达谱进行比较。

iPS细胞（ 图1A）培养在分化条 ​​件以诱导分化成的iPS-RPE细胞。上IPS-RPE展出的六角形色素细胞形态（ 图1B）相似，胎儿视网膜色素上皮（ 图1C）经典的视网膜色素上皮表型。 为了更好地理解了miRNA可能分化过程中发挥由iPS来的iPS-RPE的作用，进行miRNA表达的微阵列分析。总RNA从iPS细胞，胎儿视网膜色素上皮，和iPS-RPE使用mirVana miRNA分离试...

Watch this Scientific Journal Video about MicroRNA Expression Profiles of Human iPS Cells, Retinal Pigment Epithelium Derived From iPS, and Fetal Retinal Pigment Epithelium at JoVE.com

MicroRNA Expression Profiles of Human iPS Cells, Retinal Pigment Epithelium Derived From iPS, and Fetal Retinal Pigment Epithelium

微小RNA（miRNA）的由人诱导多能干细胞（iPS）细胞（iPS-RPE）和胎儿视网膜色素上皮衍生的人诱导多能干细胞的访问（iPS细胞），视网膜色素上皮细胞（RPE），进行比较。

人类iPS细胞的微小RNA表达谱，视网膜色素上皮衍生自IPS和胎儿视网膜色素上皮

The objective of this report is to describe the protocols for comparing the microRNA (miRNA) profiles of human induced-pluripotent stem (iPS) cells, ...

Research

JoVE Journal

Biology

Subretinal Transplantation of Human Embryonic Stem Cell Derived-retinal Pigment Epithelial Cells into a Large-eyed Model of Geographic Atrophy

人胚胎干细胞源性视网膜色素上皮细胞移植视网膜大眼模型的地理萎缩

Geographic atrophy (GA), the late stage of dry age-related macular degeneration is characterized by loss of the retinal pigment epithelial (RPE) layer, ...

科研

生物学

Primary Cell Cultures from the Mouse Retinal Pigment Epithelium

小鼠视网膜色素上皮细胞的原代培养

The retinal pigment epithelium (RPE) is a highly polarized multi-functional epithelium that is located between the neural retina and the choroid of the ...

Efficient Dissection and Culture of Primary Mouse Retinal Pigment Epithelial Cells

原发性小鼠视网膜色素上皮细胞的有效解剖与培养

Eye disorders affect millions of people worldwide, but the limited availability of human tissues hinders their study. Mouse models are powerful tools to ...

Nitroreductase/Metronidazole-Mediated Ablation and a MATLAB Platform (RpEGEN) for Studying Regeneration of the Zebrafish Retinal Pigment Epithelium

Nitroreductase/Metronidazole-Mediated Ablation and a MATLAB Platform RpEGEN for Studying Regeneration of the Zebrafish Retinal Pigment Epithelium

硝基还原酶/甲硝唑介导的消融和MATLAB平台（RpEGEN）研究斑马鱼视网膜色素上皮的再生

The retinal pigment epithelium (RPE) resides at the back of the eye and performs functions essential for maintaining the health and integrity of adjacent ...

Primary Culture of Porcine Retinal Pigment Epithelial Cells

猪视网膜色素上皮细胞原代培养

The retinal pigment epithelium (RPE) is a monolayer of polarized pigmented epithelial cells, located between the choroid and neuroretina in the retina. ...

Isolation of Primary Mouse Retinal Pigmented Epithelium Cells

小鼠原代视网膜色素上皮细胞的分离

The retinal pigmented epithelium (RPE) layer lies immediately behind the photoreceptors and harbors a complex metabolic system that plays several critical ...

Efficient and Consistent Generation of Retinal Pigment Epithelium/Choroid Flatmounts from Human Eyes for Histological Analysis

从人眼高效、一致地生成视网膜色素上皮/脉络膜平面安装，用于组织学分析

The retinal pigment epithelium (RPE) and retina are functionally and structurally connected tissues that work together to regulate light perception and ...

Real-Time Analysis of Bioenergetics in Primary Human Retinal Pigment Epithelial Cells Using High-Resolution Respirometry

使用高分辨率呼吸测定法实时分析原代人视网膜色素上皮细胞中的生物能量学

Metabolic dysfunction of retinal pigment epithelial cells (RPE) is a key pathogenic driver of retinal diseases such as age-related macular degeneration ...