Name: development of a refined protocol for trans-scleral subretinal transplantation of human retinal pigment epithelial cells into rat eyes
Uploaded: 2017-08-12T13:00:00
Description: Watch this Scientific Journal Video about Development of a Refined Protocol for Trans-scleral Subretinal Transplantation of Human Retinal Pigment Epithelial Cells into Rat Eyes at JoVE.com

我们希望感谢帕蒂吉莱德曼在手术和苏珊. 波顿为 RPE 细胞准备的帮助。我们也承认 NYSTEM C028504 为这个项目提供资金。贾斯汀· d ·米勒得到 NIH 授予 F32EY025931 的支持。

所有涉及动物的程序都得到了纽约州立大学奥尔巴尼分校动物保育和使用委员会 (IACUC) 的批准。
1. 预注射制剂
<ol><li>
hRPESC 视网膜色素上皮细胞悬浮液的制备 注意: 所有以下步骤都在无菌组织培养罩中执行, 并且需要熟悉基本的无菌技术。<ol>
<li>从50-90 岁的人眼中分离出主要的 hRPE 细胞, 并在24井板中培养细胞12。冷冻的细胞在通道 1, 解冻的?...

<ol>
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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+agerelated+macular+degeneration.">Mechanisms of agerelated macular degeneration.</a> Neuron. 75, 26-39 (2012).</li><li>Abdelsalam, A., Del Priore, L. V., Zarbin, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drusen+in+age-related+macular+degeneration:+Pathogenesis,+natural+course,+and+laser+photocoagulation-induced+regression.">Drusen in age-related macular degeneration: Pathogenesis, natural course, and laser photocoagulation-induced regression.</a> Surv Ophthalmol. 44 (1), 1-29 (1999).</li><li>Jager, R. D., Mieler, W. F., Miller, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-related+macular+degeneration.">Age-related macular degeneration.</a> N Engl J Med. 358 (24), 2606-2617 (2008).</li><li>Lund, R. 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=Human+embryonic+stem+cell-derived+cells+rescue+visual+function+in+dystrophic+RCS+rats.">Human embryonic stem cell-derived cells rescue visual function in dystrophic RCS rats.</a> Cloning Stem Cells. 8 (3), 189-199 (2006).</li><li>Vugler, 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=Embryonic+stem+cells+and+retinal+repair.">Embryonic stem cells and retinal repair.</a> Mech Dev. 124 (11-12), 807-829 (2007).</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 (9817), 713-720 (2012).</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=Human+embryonic+stem+cell-derived+retinal+pigment+epithelium+in+patients+with+age-related+macular+degeneration+and+Stargardt's+macular+dystrophy:+follow-up+of+two+open-label+phase+1/2+studies.">Human embryonic stem cell-derived retinal pigment epithelium in patients with age-related macular degeneration and Stargardt's macular dystrophy: follow-up of two open-label phase 1/2 studies.</a> Lancet. 385 (9967), 509-516 (2015).</li><li>Stanzel, B. V., 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+RPE+Stem+Cells+Grown+into+Polarized+RPE+Monolayers+on+a+Polyester+Matrix+Are+Maintained+after+Grafting+into+Rabbit+Subretinal+Space.">Human RPE Stem Cells Grown into Polarized RPE Monolayers on a Polyester Matrix Are Maintained after Grafting into Rabbit Subretinal Space.</a> Stem Cell Reports. 2 (1), 64-77 (2014).</li><li>Blenkinsop, T. 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=Human+adult+retinal+pigment+epithelial+stem+cell-derived+RPE+monolayers+exhibit+key+physiological+characteristics+of+native+tissue.">Human adult retinal pigment epithelial stem cell-derived RPE monolayers exhibit key physiological characteristics of native tissue.</a> Invest Ophthalmol Vis Sci. 56 (12), 7085-7099 (2015).</li><li>Salero, 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=Adult+human+RPE+can+be+activated+into+a+multipotent+stem+cell+that+produces+mesenchymal+derivatives.">Adult human RPE can be activated into a multipotent stem cell that produces mesenchymal derivatives.</a> Cell Stem Cell. 10 (1), 88-95 (2012).</li><li>Davis, J. 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=Human+RPE+Stem+Cell-Derived+RPE+Preserves+Photoreceptors+in+the+Royal+College+of+Surgeons+Rat:+Method+for+Quantifying+the+Area+of+Photoreceptor+Sparing.">Human RPE Stem Cell-Derived RPE Preserves Photoreceptors in the Royal College of Surgeons Rat: Method for Quantifying the Area of Photoreceptor Sparing.</a> Journal of Ocular Pharmacology and Therapeutics. 32 (5), 304-309 (2016).</li><li>Westenskow, P. 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=Performing+Subretinal+Injections+in+Rodents+to+Deliver+Retinal+Pigment+Epithelium+Cells+in+Suspension.">Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension.</a> J Vis Exp. (95), e52247 (2015).</li><li>Lopez, 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=Transplanted+Retinal+Pigment+Epithelium+Modifies+the+Retinal+Degeneration+in+the+RCS+Rat.">Transplanted Retinal Pigment Epithelium Modifies the Retinal Degeneration in the RCS Rat.</a> Invest Ophthalmol Vis Sci. 30 (3), 586-588 (1989).</li><li>Eberle, D., Santos-Ferreira, T., Grahl, S., Ader, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subretinal+Transplantation+of+MACS+Purified+Photoreceptor+Precursor+Cells+into+the+Adult+Mouse+Retina.">Subretinal Transplantation of MACS Purified Photoreceptor Precursor Cells into the Adult Mouse Retina.</a> J Vis Exp. (84), e50932 (2014).</li><li>Nair, 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=Effects+of+Common+Anesthetics+on+Eye+Movement+and+Electroretinogram.">Effects of Common Anesthetics on Eye Movement and Electroretinogram.</a> Doc Ophthalmol. 122 (3), 163-176 (2011).</li><li>McGill, T. 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=Transplantation+of+human+central+nervous+system+stem+cells+-+neuroprotection+in+retinal+degeneration.">Transplantation of human central nervous system stem cells - neuroprotection in retinal degeneration.</a> Eur J Neurosci. 35, 468-477 (2012).</li><li>Al-Hussaini, H., Kam, J. H., Vugler, A., Semo, M., Jeffery, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mature+retinal+pigment+epithelium+cells+are+retained+in+the+cell+cycle+and+proliferate+in+vivo.">Mature retinal pigment epithelium cells are retained in the cell cycle and proliferate in vivo.</a> Mol Vis. 14, 1784-1791 (2008).</li><li>Wang, S., Lu, B., Wood, P., Lund, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Grafting+of+ARPE-19+and+Schwann+Cells+to+the+Subretinal+Space+in+RCS+Rats.">Grafting of ARPE-19 and Schwann Cells to the Subretinal Space in RCS Rats.</a> Invest Ophthalmol Vis Sci. 46 (7), 2552-2560 (2005).</li><li>Fabian, R. J., Bond, J. M., Drobeck, H. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induced+corneal+opacities+in+the+rat.">Induced corneal opacities in the rat.</a> Br J Ophthalmol. 51 (2), 124-129 (1967).</li></ol>

本文所描述的视网膜注射技术是通过跨巩膜通路, 注射器针穿透眼壁的外层 (巩膜-脉络膜-RPE 复合体) 而不损害神经视网膜或扰乱玻璃腔。一个替代的反式 vitreal 方法有潜在的风险, 导致白内障的晶状体损伤, 因为啮齿目动物的镜头占据了大多数的玻璃腔。与这种方法相比, 我们的技术是低风险和造成最小的创伤, 因为注射器针不需要穿过整个玻璃腔, 以达到视网膜空间。事实上, 在我们的研究中, OCT ?...

人视网膜位于眼睛的后面功能作为一个轻的感觉组织并且在视觉知觉扮演一个关键的角色。因此视网膜细胞功能障碍或细胞死亡导致视力问题或永久性失明。在视网膜不同层次的细胞变性或功能紊乱的疾病被称为退行性视网膜疾病, 其中 AMD 是最常见的类型, 也是发达国家老年人不可逆转性失明的主要原因之一1,2。AMD 的病理过程与视网膜色素上皮层与底层赫膜之间的 "疣" 堆积有关, 从而损害了视网膜色素上皮支持的感光生理, 导致了神经系统的萎缩和视力丧失3, 4,5。到目前为止, 没有治疗高级干 (non-neovascular) AMD。干细胞治疗作为再生医学的一个新范式的出现, 带来了用干细胞衍生的健康细胞取代功能失调或死亡的 RPE 细胞的希望。事实上, 广泛的临床前研究的移植干细胞 (如, 人类胚胎干细胞) 衍生 rpe 细胞到 rpe-变性动物模型已经执行6,7, 其中一些已取得进展临床试验8,9 (NCT01344993, ClinicalTrials.gov)。最近, 我们的实验室发现了人类视网膜色素上皮细胞 (hRPESCs) 中的一种替代的干细胞来源, 目前正被用于 hRPESC 衍生-rpe 细胞 (hRPESC-rpe) 移植治疗 AMD 的前临床研究。10,11,12,13。
视网膜注射技术应用于上述多个群体, 包括本组的前临床研究。动物视网膜注射有两种一般的方法: 反 vitreal 和反式巩膜。vitreal 方法的优势是, 外科医生能够直接观察针端, 因为它穿透了前眼, 越过整个 vitreal 腔毗邻的镜头, 并穿透视网膜在后面的眼睛, 以达到视网膜空间14,15,16。然而, 它需要扰乱视网膜在两个位置 (前部和后部), 承担损害晶状体的风险, 并可能导致细胞回流到玻璃体时, 针被收回。相反, 跨巩膜的方法, 原则上, 避免介入视网膜和玻璃体, 和回流退出眼睛。在有色啮齿目动物中, 外科医生最初可以观察到巩膜的穿透, 但在进入色素脉络后, 针端不再可见。没有直接观察, 破坏视网膜是常见的, 并可能导致视网膜解剖和交付细胞和/或血液进入玻璃体。此外, 由于眼睛表面是弯曲的, 很难知道哪种针的角度和深度是最有效的反式巩膜注射。
在这一可视化的文章中, 我们介绍了一个跨巩膜视网膜注射方法, 通过使用后评价与光学相干断层扫描 (OCT), 这使得对注射场的详细检查。我们的反式巩膜注射技术利用定义的位置, 角度和深度注射针, 以产生非常低的外科创伤和高可靠性。在这里, 我们特别演示了 hRPESC-RPE 细胞注入视网膜空间的 RCS 大鼠, 一个临床前模型的人 AMD。采用这种注射方法, 成功、一致地将 hRPESC 视网膜色素上皮细胞导入到 RCS 大鼠眼的视网膜空间中, 成功率很高。注入的细胞以前发现, 导致保存 RCS 光至少2月后, 注入13。本程序在解剖显微镜下进行, 易于学习。它需要两个人 (一个外科医生和一个助理) 来执行注射和平均注射时间为每种动物少于5分钟。定义的角度和深度为注射针, 使之成为可能的实验室, 在那里 OCT 是无法获得成功的视网膜注射液。它允许高度可再生的视网膜通路, 不仅可以用于细胞移植, 也可用于药物传递和基因治疗。

<table border="0" cellpadding="0" cellspacing="0" width="863">
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			<td height="21" style="height:21px;width:259px;">0.25% Trypsin-EDTA (1x)</td>
			<td style="width:256px;">Life Technologies</td>
			<td style="width:119px;">25200-072</td>
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			<td height="21" style="height:21px;width:259px;">DNAse I</td>
			<td style="width:256px;">Sigma</td>
			<td style="width:119px;">DN-25</td>
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			<td height="63" style="height:63px;width:259px;">1xDulbecco&#8217;s Phosphate Buffered Saline without Calcium &#38; Magnesium (1xDPBS-CMF)</td>
			<td style="width:256px;">Corning Cellgro</td>
			<td style="width:119px;">431219</td>
			<td style="width:231px;"></td>
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			<td height="21" style="height:21px;width:259px;">Sterile Balanced Salt Solution (BSS)</td>
			<td style="width:256px;">Alcon</td>
			<td style="width:119px;">00065079550</td>
			<td style="width:231px;"></td>
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			<td height="21" style="height:21px;width:259px;">Sterile eye wash</td>
			<td style="width:256px;">Moore Medical</td>
			<td style="width:119px;">75519</td>
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			<td height="21" style="height:21px;width:259px;">Sterile 0.9% saline</td>
			<td style="width:256px;">Hospira</td>
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			<td height="42" style="height:42px;width:259px;">Proparacaine Hydrochloride Ophthalmic Solution (0.5%)</td>
			<td style="width:256px;">Akorn</td>
			<td style="width:119px;">17478026312</td>
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			<td height="42" style="height:42px;width:259px;">Tropicamide Ophthalmic Solution, USP (1%)</td>
			<td style="width:256px;">Bausch &#38; Lomb</td>
			<td style="width:119px;">24208058559</td>
			<td style="width:231px;"></td>
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			<td height="42" style="height:42px;width:259px;">Phenylepherine Ophtalmic Solution, USP (10%) stock</td>
			<td style="width:256px;">Bausch &#38; Lomb</td>
			<td style="width:119px;">42702010305</td>
			<td style="width:231px;">This is used to make 2.5% Phenylepherine</td>
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			<td height="21" style="height:21px;width:259px;">Buprenex</td>
			<td style="width:256px;">Patterson</td>
			<td style="width:119px;">433502</td>
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			<td height="21" style="height:21px;width:259px;">Dexamethasone</td>
			<td style="width:256px;">APP Pharmaceuticals</td>
			<td style="width:119px;">63323051610</td>
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			<td height="21" style="height:21px;width:259px;">100% Ethanol</td>
			<td style="width:256px;">Thermo Scientific</td>
			<td style="width:119px;">615090040</td>
			<td style="width:231px;"></td>
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			<td height="21" style="height:21px;width:259px;">70% Ethanol</td>
			<td style="width:256px;">Ricca Chemical Company</td>
			<td style="width:119px;">2546.70-5</td>
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			<td height="24" style="height:24px;width:259px;">Sterile GenTeal Lubricant Eye Gel</td>
			<td style="width:256px;">Novartis</td>
			<td style="width:119px;">78042947</td>
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			<td height="42" style="height:42px;width:259px;">Sterile Systane Ultra Lubricant Eye Drops</td>
			<td style="width:256px;">Alcon</td>
			<td style="width:119px;">00065143105</td>
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			<td height="44" style="height:44px;width:259px;">hRPESC-RPE cells</td>
			<td style="width:256px;">Not available commercially</td>
			<td style="width:119px;"></td>
			<td style="width:231px;">Please refer to &#34;Reference #12&#34; for cell isolation and mainteinance.</td>
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			<td height="21" style="height:21px;width:259px;">24-well plates</td>
			<td style="width:256px;">Corning</td>
			<td style="width:119px;">3526</td>
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			<td height="21" style="height:21px;width:259px;">Conical tubes (15 ml)</td>
			<td style="width:256px;">Sarstedt</td>
			<td style="width:119px;">62554002</td>
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			<td height="21" style="height:21px;width:259px;">Microcentrifuge cap with o-ring</td>
			<td style="width:256px;">LPS inc</td>
			<td style="width:119px;">L233126</td>
			<td style="width:231px;"></td>
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			<td height="22" style="height:22px;width:259px;">Capless Microcentrifuge tubes (1.7 ml)</td>
			<td style="width:256px;">LPS inc</td>
			<td style="width:119px;">L233041</td>
			<td style="width:231px;"></td>
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			<td height="21" style="height:21px;width:259px;">Centrifuge</td>
			<td style="width:256px;">Eppendorf</td>
			<td style="width:119px;">5804R</td>
			<td style="width:231px;"></td>
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			<td height="21" style="height:21px;width:259px;">Sterile alcohol wipe</td>
			<td style="width:256px;">McKesson</td>
			<td style="width:119px;">58-204</td>
			<td style="width:231px;"></td>
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			<td height="21" style="height:21px;width:259px;">Sterile cotton tip applicators</td>
			<td style="width:256px;">McKesson</td>
			<td style="width:119px;">24-106-2S</td>
			<td style="width:231px;"></td>
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			<td height="21" style="height:21px;width:259px;">Sterile Weck-Cel spears</td>
			<td style="width:256px;">Beaver-Visitec International&#160;</td>
			<td style="width:119px;">0008680</td>
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			<td height="21" style="height:21px;width:259px;">Sterile surgical drapes&#160;</td>
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development of a refined protocol for trans-scleral subretinal transplantation of human retinal pigment epithelial cells into rat eyes

退行性视网膜疾病, 如年龄相关的黄斑变性 (AMD) 是导致不可逆转的视力丧失全球。AMD 的特点是视网膜色素上皮 (RPE) 细胞的退化, 这是一个单层的细胞功能支持和解剖包裹周围的神经视网膜。目前的药理治疗 non-neovascular amd (干 amd) 只减缓疾病进展, 但不能恢复视力, 必须研究, 以确定新的治疗策略。用健康细胞取代退行性视网膜色素上皮细胞有望在未来治疗干性 AMD。广泛的临床前研究的干细胞替代疗法的 AMD 涉及移植干细胞衍生 RPE 细胞进入视网膜空间的动物模型, 其中视网膜注射技术的应用。在这些临床前动物研究中最常用的方法是通过经巩膜路线, 这是由于缺乏直接可视化的针端, 往往会导致视网膜损伤而变得困难。通过玻璃体的另一种方法可以直接观察针端位置 , 但是由于更多的眼部组织扰 , 它有很高的手术创伤风险。我们已经开发了一个低风险和可重复性的改良反式巩膜注射法, 使用定义的针角和深度, 成功地和一贯地提供 RPE 细胞进入大鼠视网膜空间, 并避免过多的视网膜损伤。以这种方式传送的细胞在皇家外科医生 (RCS) 大鼠中已经被证明是有效的, 至少2月。该技术不仅可用于细胞移植, 也可用于小分子或基因治疗的传递。

利用本文所描述的技术, 我们通过精确地控制插入到组织中的注射器针的位置、角度和深度, 将 hRPESC 视网膜色素上皮细胞送到 RCS 大鼠的视网膜空间中 (图 1B-D).在移植后立即进行 OCT 检查, 观察注射部位和移植细胞产生的视网膜疱疹。术后 OCT 评估是评估注射质量和视网膜损伤或出血监测的检查工具。在 OCT 扫描下, 可以清楚地看到视网膜疱疹 (<strong...

Watch this Scientific Journal Video about Development of a Refined Protocol for Trans-scleral Subretinal Transplantation of Human Retinal Pigment Epithelial Cells into Rat Eyes at JoVE.com

Development of a Refined Protocol for Trans-scleral Subretinal Transplantation of Human Retinal Pigment Epithelial Cells into Rat Eyes

视网膜注射液已广泛应用于干细胞置换治疗老年性黄斑变性的临床前研究。在这一可视化的文章中, 我们描述了一个低风险, 可重复和精确修改的视网膜注射技术, 通过巩膜的方法传递细胞到大鼠的眼睛。

人视网膜色素上皮细胞向大鼠眼巩膜视网膜移植精制方案的研制

Degenerative retinal diseases such as age-related macular degeneration (AMD) are the leading cause of irreversible vision loss worldwide. AMD is ...

Research

JoVE Journal

Neuroscience

Experimental Models for Study of Retinal Pigment Epithelial Physiology and Pathophysiology

视网膜色素上皮的生理与病理生理学研究的实验模型

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium ...

科研

神经科学

Dissection of a Mouse Eye for a Whole Mount of the Retinal Pigment Epithelium

解剖鼠标眼的视网膜色素上皮的全山

The retinal pigment epithelium (RPE) lies at the back of the mammalian eye, just under the neural retina, which contains the photoreceptors (rods and ...

Isolation and Culture of Rat Embryonic Neural Cells: A Quick Protocol

大鼠胚胎神经细胞的分离和培养：一个快速议定书“

We are describing a quick method to dissociate and culture hippocampal or cortical neurons from E15-17 rat embryos. The procedure can be applied ...

Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium

病毒载体的基因治疗的小鼠视网膜色素上皮缘方针，视网膜下注射

The eye is a small and enclosed organ which makes it an ideal target for gene therapy. Recently various strategies have been applied to gene therapy in ...

Enrichment of Bruch's Membrane from Human Donor Eyes

从人类供体眼富集布鲁赫的膜

Age-related macular degeneration (AMD) is a leading cause of visual impairment in the developed world. The disease manifests itself by the destruction of ...

Single-cell RNA-Seq of Defined Subsets of Retinal Ganglion Cells

视网膜神经节细胞定义子集的单细胞RNA-Seq

The discovery of cell type-specific markers can provide insight into cellular function and the origins of cellular heterogeneity. With a recent push for ...

A Protocol for Measuring Cue Reactivity in a Rat Model of Cocaine Use Disorder

可卡因使用障碍大鼠模型中提示反应性测定的一种协议

Cocaine use disorder (CUD) follows a trajectory of repetitive self-administration during which previously neutral stimuli gain incentive value. Cue ...

Homochronic Transplantation of Interneuron Precursors into Early Postnatal Mouse Brains

Interneuron 前体 Homochronic 移植早期小鼠脑的研究

Neuronal fate determination and maturation requires an intricate interplay between genetic programs and environmental signals. However, disentangling the ...

Transplantation of Chemogenetically Engineered Cortical Interneuron Progenitors into Early Postnatal Mouse Brains

将化学遗传工程皮质内神经内生前生移植到早期产后小鼠大脑

Neuronal development is regulated by a complex combination of environmental and genetic factors. Assessing the relative contribution of each component is ...

3D Kinematic Analysis for the Functional Evaluation in the Rat Model of Sciatic Nerve Crush Injury

坐骨神经粉碎损伤大鼠模型中功能评估的3D运动学分析

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of ...