Name: subretinal injection of gene therapy vectors and stem cells in the perinatal mouse eye
Uploaded: 2012-11-25T14:00:00
Description: Watch this Scientific Journal Video about Subretinal Injection of Gene Therapy Vectors and Stem Cells in the Perinatal Mouse Eye at JoVE.com

研究以预防失明实验孝行长崎的协助下，这项研究符合眼科及视觉研究中使用动物的ARVO声明。 KJW支持由美国国立卫生研究院资助5T32EY013933和5T32DK007647-20。 VBM是由美​​国国立卫生研究院授予K08EY020530支持。

1。视网膜下注射组装设备<ol><li>购买或制作100微米直径的针从一个玻璃毛细管。这可以手动完成使用萨特P-97的移液管的牵拉或其他类似设备。将被加热的毛细管的端部和拉出，直到达到所希望的直径（100微米）。的较小直径的针可用于基因治疗的载体，但是，这是无损伤的细胞或眼睛的推荐直径为细胞注射。玻璃毛细管针钢针相比，具有非常细的，但钝头，用于可视化的喷射流体和访问到视网...

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	<li>Abe, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+of+the+retina+using+pigment+epithelial+cell+transplantation.">Regeneration of the retina using pigment epithelial cell transplantation.</a> Nihon Ganka Gakkai Zasshi. 106, 778-803 (2002).</li><li>Jacobson, S. G., Cideciyan, A. V., Ratnakaram, R., Heon, E., Schwartz, S. B., Roman, A. J., Peden, M. C., Aleman, T. S., Boye, S. L., Sumaroka, 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=Gene+therapy+for+Leber+congenital+amaurosis+caused+by+RPE65+mutations:+safety+and+efficacy+in+15+children+and+adults+followed+up+to+3+years.">Gene therapy for Leber congenital amaurosis caused by RPE65 mutations: safety and efficacy in 15 children and adults followed up to 3 years.</a> Arch. Ophthalmol. 130, 9-24 (2011).</li><li>Schwartz, S. D., Hubschman, J. P., Heilwell, G., Franco-Cardenas, V., Pan, C. K., Ostrick, R. M., Mickunas, E., Gay, R., Klimanskaya, I., Lanza, R. <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>Wang, N. K., Tosi, J., Kasanuki, J. M., Chou, C. L., Kong, J., Parmalee, N., Wert, K. J., Allikmets, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+reprogrammed+embryonic+stem+cells+improves+visual+function+in+a+mouse+model+for+retinitis+pigmentosa.">Transplantation of reprogrammed embryonic stem cells improves visual function in a mouse model for retinitis pigmentosa.</a> Transplantation. 89, 911-919 (2010).</li><li>Song, B. J., Tsang, S. H., Lin, C. -. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+models+of+retinal+degeneration+and+targets+for+gene+therapy.">Genetic models of retinal degeneration and targets for gene therapy.</a> Gene Ther. Mol. Biol. 11, 229-262 (2007).</li><li>Tosi, J., Sancho-Pelluz, J., Davis, R. J., Hsu, C. W., Wolpert, K. V., Sengillo, J. D., Lin, C. S., Tsang, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lentivirus-mediated+expression+of+cDNA+and+shRNA+slows+degeneration+in+retinitis+pigmentosa.">Lentivirus-mediated expression of cDNA and shRNA slows degeneration in retinitis pigmentosa.</a> Exp. Biol. Med. 236, 1211-1217 (2011).</li><li>Tucker, B. A., Park, I. H., Qi, S. D., Klassen, H. J., Jiang, C., Yao, J., Redenti, S., Daley, G. Q., Young, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+adult+mouse+iPS+cell-derived+photoreceptor+precursors+restores+retinal+structure+and+function+in+degenerative+mice.">Transplantation of adult mouse iPS cell-derived photoreceptor precursors restores retinal structure and function in degenerative mice.</a> PLoS One. 6, e189992 (2011).</li><li>Mahajan, V. B., Mondino, B. J., Tsang, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> A high-throughput Mouse Eye Phenomics System. , (2010).</li><li>Chang, B., Hawes, N. L., Hurd, R. E., Davisson, M. T., Nusinowitz, S., Heckenlively, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinal+degeneration+mutants+in+the+mouse.">Retinal degeneration mutants in the mouse.</a> Vision Res. 42, 517-525 (2002).</li><li>Hawes, N. L., Smith, R. S., Chang, B., Davisson, M., Heckenlively, J. R., John, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+fundus+photography+and+angiography:+a+catalogue+of+normal+and+mutant+phenotypes.">Mouse fundus photography and angiography: a catalogue of normal and mutant phenotypes.</a> Mol. Vis. 5, 22 (1999).</li><li>Won, J., Shi, L. Y., Hicks, W., Wang, J., Hurd, R., Naggert, J. K., Chang, B., Nishina, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+model+resources+for+vision+research.">Mouse model resources for vision research.</a> J. Ophthalmol. 2011, 391384 (2011).</li></ol>

此视频技术提供视网膜下注射外科手术成功完成，并且确保被放置在所需的位置，有效地治疗眼科疾病的基因治疗载体或干细胞的说明。这种技术允许RPE或光感受器如视网膜细胞的定位，因为它的基因治疗载体或干细胞衍生的组织放置在这些细胞的周边。以前的方法，其中流体被放置在玻璃体腔的玻璃体内注射，而且必须通过视网膜移植，以针对这些特定领域。玻璃体内注射可降低转导的基因治?...

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 目录 </td><td> 评论 </td></tr><tr><td> 0.8-1.10×100毫米的毛细管（玻璃） </td><td> ，金布尔玻璃公司</td><td> 34502 99 </td><td></td></tr><tr><td>烈焰红唇/褐色微管牵引机</td><td>萨特仪器</td><td> P-97 </td><td>成茂microforge可以用来代替。产品编号MF-900 </td></tr><tr><td> Sigmacote </td><td> Sigma Aldrich公司</td><td> SL2-25ML </td><td>有机硅</td></tr><tr><td> Dubecco的磷酸盐缓冲液，氯化钙和氯化镁</td><td> GIBCO-Invitrogen公司</td><td> 14040-133 </td><td></td></tr><tr><td>安全乐25 3/4G×12“;采血车集</td><td> BD真空采血管</td><td> 367298 </td><td></td></tr><tr><td> 1毫升子-Q 265/8G防滑提示注射器</td><td>贝克顿 - 迪金森</td><td> 309597 </td><td></td></tr><tr><td> 0.5-10微升Finnpipette II音量可调移液器</td><td> Fisherbrand </td><td> 21-377-815 </td><td></td></tr><tr><td> 1-200微升自然斜角提示</td><td>美国科技公司</td><td> 1111至1700年</td><td></td></tr><tr><td> Discovery.V8体视显微镜</td><td>蔡司</td><td> MC1500 </td><td></td></tr><tr><td> 60毫米×15毫米风格处理聚苯乙烯细胞培养皿</td><td>康宁公司</td><td> 430166 </td><td></td></tr><tr><td> Vannas直剪刀</td><td> STORZ眼药水</td><td> E3383小号</td><td></td></tr><tr><td>弯曲的敷料钳锯齿精致</td><td> STORZ眼药水</td><td> E1408 </td><td></td></tr><tr><td> 15度显微刀</td><td>厚朴眼科公司</td><td> 091204 </td><td></td></tr><tr><td>氯胺酮</td><td> Ketaset III </td><td> NADA＃45-290 </td><td></td></tr><tr><td>二甲苯胺噻嗪</td><td>劳埃德实验室</td><td> NADA＃139-236 </td><td></td></tr><tr><td>布比卡因（腔投与局部麻醉药 - 麻佳因） </td><td>阿斯利康公司</td><td> N / A </td><td></td></tr><tr><td>丁丙诺啡</td><td> Sigma Aldrich公司</td><td> B9275 </td><td></td></tr></tbody></table>

subretinal injection of gene therapy vectors and stem cells in the perinatal mouse eye

视力丧失，影响约3.4万人在美国，在未来数年预计将增加。1，基因治疗和干细胞移植已成为主要的治疗手段，治疗视网膜退行性疾病导致的失明。几种形式的自体移植的年龄相关性黄斑变性（AMD），如虹膜色素上皮细胞移植，已经产生了令人鼓舞的结果，为其他形式的基因和干细胞疗法的人体临床试验已经开始。2这些包括RPE65基因置换Leber氏先天性黑蒙和Stargardt病利用人类胚胎干（ES）细胞中的RPE细胞移植治疗的患者。3-4现在有基因治疗的载体，可用于治疗视网膜疾病的患者的干细胞，它是重要的，以验证这些潜在的治疗在动物模型中，前申请ING他们在人类研究中。该鼠标已经成为一个重要的科学模型测试基因治疗的载体和造血干细胞移植在眼的疗效。5-8在这个视频文章中，我们提出了一个技术基因治疗的载体或干细胞分化为视网膜下腔注入老鼠的眼睛，同时最大限度地减少向周围组织的损害。

一个绘图作参考标记的主要结构，示出的鼠标眼玻璃体腔和视网膜下注射的外科手术（箭头， 见图1）的位置显示的箭头。基因治疗的载体，如的LacZ基因的慢病毒载体（ 图2），可以使用这些位置注入。另外，骨髓间充质干细胞，如小鼠胚胎干细胞（ 图3），也可以在这些位点中的老鼠的眼睛被移植。 1微升由巨细胞病毒（CMV）启?...

Watch this Scientific Journal Video about Subretinal Injection of Gene Therapy Vectors and Stem Cells in the Perinatal Mouse Eye at JoVE.com

Subretinal Injection of Gene Therapy Vectors and Stem Cells in the Perinatal Mouse Eye

这种手术的方法说明了基因治疗的载体和干细胞注射到老鼠的眼睛的视网膜下腔。

基因治疗的载体和干细胞视网膜下注射到老鼠的眼睛

The loss of sight affects approximately 3.4 million people in the United States and is expected to increase in the upcoming years.1 Recently, gene therapy ...

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在癌症的生产和检测活性氧（ROS）的

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