Name: using a cell-tracer injection to investigate the origin of neointima-forming cells in a rat saccular side wall model
Uploaded: 2022-03-16T14:45:00
Description: Watch this Scientific Journal Video about Using a Cell-Tracer Injection to Investigate the Origin of Neointima-Forming Cells in a Rat Saccular Side Wall Model at JoVE.com

作者感谢Alessandra Bergadano，DVM，PhD，对长期动物健康的专门监督。这项工作得到了研究委员会，瑞士阿劳Kantonsspital Aarau和瑞士国家科学基金会SNF（310030_182450）的研究基金的支持。

兽医支持是根据机构指南进行的。实验由瑞士地方伦理委员会批准（BE 60/19）。6，7严格遵守了ARRIAGE指南和3R原则。包括31只雄性刘易斯大鼠，12周大，体重492±8克。将所有大鼠在23°C的室温和12小时的光/暗循环下。提供免费水和颗粒。统计分析是使用非参数Wilcoxon-Mann-Whitney U 检验进行的。≤0.05和/或≤0.01的概率值（p）被认为是显著的。
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<ol>
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L., 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=Preventing+vasospasm+improves+outcome+after+aneurysmal+subarachnoid+hemorrhage:+rationale+and+design+of+CONSCIOUS-2+and+CONSCIOUS-3+trials.">Preventing vasospasm improves outcome after aneurysmal subarachnoid hemorrhage: rationale and design of CONSCIOUS-2 and CONSCIOUS-3 trials.</a> Neurocritical Care. 13 (3), 416-424 (2010).</li><li>Wanderer, 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=Levosimendan+as+a+therapeutic+strategy+to+prevent+neuroinflammation+after+aneurysmal+subarachnoid+hemorrhage.">Levosimendan as a therapeutic strategy to prevent neuroinflammation after aneurysmal subarachnoid hemorrhage.</a> Journal of Neurointerventional Surgery. , (2021).</li><li>Wanderer, 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=Aspirin+treatment+prevents+inflammation+in+experimental+bifurcation+aneurysms+in+New+Zealand+White+rabbits.">Aspirin treatment prevents inflammation in experimental bifurcation aneurysms in New Zealand White rabbits.</a> Journal of Neurointerventional Surgery. 14 (2), 189-195 (2021).</li><li>Gruter, B. 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=Patterns+of+neointima+formation+after+coil+or+stent+treatment+in+a+rat+saccular+sidewall+aneurysm+model.">Patterns of neointima formation after coil or stent treatment in a rat saccular sidewall aneurysm model.</a> Stroke. 52 (3), 1043-1052 (2021).</li><li>Kilkenny, C., 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=Animal+research:+reporting+in+vivo+experiments:+the+ARRIVE+guidelines.">Animal research: reporting in vivo experiments: the ARRIVE guidelines.</a> British Journal of Pharmacology. 160 (7), 1577-1579 (2010).</li><li>Tornqvist, 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=Strategic+focus+on+3R+principles+reveals+major+reductions+in+the+use+of+animals+in+pharmaceutical+toxicity+testing.">Strategic focus on 3R principles reveals major reductions in the use of animals in pharmaceutical toxicity testing.</a> PLoS One. 9 (7), 101638 (2014).</li><li>Nevzati, 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=Aneurysm+wall+cellularity+affects+healing+after+coil+embolization:+assessment+in+a+rat+saccular+aneurysm+model.">Aneurysm wall cellularity affects healing after coil embolization: assessment in a rat saccular aneurysm model.</a> Journal of Neurointerventional Surgery. 12 (6), 621-625 (2020).</li><li>Marbacher, 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=The+Helsinki+rat+microsurgical+sidewall+aneurysm+model.">The Helsinki rat microsurgical sidewall aneurysm model.</a> Journal of Visualized Experiments: JoVE. (92), e51071 (2014).</li><li>Nevzati, 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=Biodegradable+magnesium+stent+treatment+of+saccular+aneurysms+in+a+rt+model+-+introduction+of+the+surgical+technique.">Biodegradable magnesium stent treatment of saccular aneurysms in a rt model - introduction of the surgical technique.</a> Journal of Visualized Experiments: JoVE. (128), e56359 (2017).</li><li>Gruter, B. 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=Testing+bioresorbable+stent+feasibility+in+a+rat+aneurysm+model.">Testing bioresorbable stent feasibility in a rat aneurysm model.</a> Journal of Neurointerventional Surgery. 11 (10), 1050-1054 (2019).</li><li>Kadirvel, 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=Cellular+mechanisms+of+aneurysm+occlusion+after+treatment+with+a+flow+diverter.">Cellular mechanisms of aneurysm occlusion after treatment with a flow diverter.</a> Radiology. 270 (2), 394-399 (2014).</li><li>Li, Z. F., 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=Endothelial+progenitor+cells+contribute+to+neointima+formation+in+rabbit+elastase-induced+aneurysm+after+flow+diverter+treatment.">Endothelial progenitor cells contribute to neointima formation in rabbit elastase-induced aneurysm after flow diverter treatment.</a> CNS Neuroscience &amp; Therapeutics. 19 (5), 352-357 (2013).</li><li>Marbacher, 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=Intraluminal+cell+transplantation+prevents+growth+and+rupture+in+a+model+of+rupture-prone+saccular+aneurysms.">Intraluminal cell transplantation prevents growth and rupture in a model of rupture-prone saccular aneurysms.</a> Stroke. 45 (12), 3684-3690 (2014).</li><li>Frosen, 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=Contribution+of+mural+and+bone+marrow-derived+neointimal+cells+to+thrombus+organization+and+wall+remodeling+in+a+microsurgical+murine+saccular+aneurysm+model.">Contribution of mural and bone marrow-derived neointimal cells to thrombus organization and wall remodeling in a microsurgical murine saccular aneurysm model.</a> Neurosurgery. 58 (5), 936-944 (2006).</li><li>Marbacher, S., Niemela, M., Hernesniemi, J., Frosen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recurrence+of+endovascularly+and+microsurgically+treated+intracranial+aneurysms-review+of+the+putative+role+of+aneurysm+wall+biology.">Recurrence of endovascularly and microsurgically treated intracranial aneurysms-review of the putative role of aneurysm wall biology.</a> Neurosurgical Review. 42 (1), 49-58 (2019).</li><li>Frosen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Smooth+muscle+cells+and+the+formation,+degeneration,+and+rupture+of+saccular+intracranial+aneurysm+wall--a+review+of+current+pathophysiological+knowledge.">Smooth muscle cells and the formation, degeneration, and rupture of saccular intracranial aneurysm wall--a review of current pathophysiological knowledge.</a> Translational Stroke Research. 5 (3), 347-356 (2014).</li><li>Fang, 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=Bone+marrow-derived+endothelial+progenitor+cells+are+involved+in+aneurysm+repair+in+rabbits.">Bone marrow-derived endothelial progenitor cells are involved in aneurysm repair in rabbits.</a> Journal of Clinical Neuroscience. 19 (9), 1283-1286 (2012).</li><li>Morel, 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=Sex-related+differences+in+wall+remodeling+and+intraluminal+thrombus+resolution+in+a+rat+saccular+aneurysm+model.">Sex-related differences in wall remodeling and intraluminal thrombus resolution in a rat saccular aneurysm model.</a> Journal of Neurosurgery. , 1-14 (2019).</li><li>Gruter, B. 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=Fluorescence+video+angiography+for+evaluation+of+dynamic+perfusion+status+in+an+aneurysm+preclinical+experimental+setting.">Fluorescence video angiography for evaluation of dynamic perfusion status in an aneurysm preclinical experimental setting.</a> Operative Neurosurgery. 17 (4), 432-438 (2019).</li><li>Marbacher, S., Strange, F., Frosen, J., Fandino, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+extracranial+aneurysm+models+for+the+study+and+treatment+of+brain+aneurysms:+A+systematic+review.">Preclinical extracranial aneurysm models for the study and treatment of brain aneurysms: A systematic review.</a> Journal of Cerebral Blood Flow &amp; Metabolism. 40 (5), 922-938 (2020).</li><li>Ravindran, 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=Mechanism+of+action+and+biology+of+flow+diverters+in+the+treatment+of+intracranial+aneurysms.">Mechanism of action and biology of flow diverters in the treatment of intracranial aneurysms.</a> Neurosurgery. 86, 13-19 (2020).</li><li>Marbacher, 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=Loss+of+mural+cells+leads+to+wall+degeneration,+aneurysm+growth,+and+eventual+rupture+in+a+rat+aneurysm+model.">Loss of mural cells leads to wall degeneration, aneurysm growth, and eventual rupture in a rat aneurysm model.</a> Stroke. 45 (1), 248-254 (2014).</li><li>Morosanu, C. O., 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=Neurosurgical+cadaveric+and+in+vivo+large+animal+training+models+for+cranial+and+spinal+approaches+and+techniques+-+systematic+review+of+current+literature.">Neurosurgical cadaveric and in vivo large animal training models for cranial and spinal approaches and techniques - systematic review of current literature.</a> Neurologia i Neurochirurgia Polska. 53 (1), 8-17 (2019).</li><li>Wanderer, 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=Arterial+pouch+microsurgical+bifurcation+aneurysm+model+in+the+rabbit.">Arterial pouch microsurgical bifurcation aneurysm model in the rabbit.</a> Journal of Visualized Experiments: JoVE. (159), e61157 (2020).</li></ol>

这项研究表明，新血管的形成是通过起源于动脉瘤复合体母动脉的内皮细胞介导的，但由重要动脉瘤中来自动脉瘤壁的细胞的募集来支持。然而，循环祖细胞在动脉瘤愈合中的作用仍然存在争议12，13。总体而言，31只雄性刘易斯大鼠被纳入这项调查;只有4人过早死亡（死亡率为12.9%）。
与促进随后内皮至内皮接触的手术夹闭相比，?...

由颅内动脉瘤 （IA） 破裂引起的蛛网膜下腔出血是一种破坏性的神经外科疾病，与高发病率和死亡率相关1，2，3，4。除了提供内皮与内皮直接接触的显微外科夹闭外，血管内装置在过去几十年中在治疗破裂和偶然发现的IA方面变得越来越重要。血管内治疗的IA的愈合反应主要取决于新脑形成和血栓组织。两者都是协同过程，取决于细胞从相邻血管和动脉瘤壁的迁移。5 迄今为止，血管内处理动脉瘤新脑形成中内皮细胞的起源尚不清楚。文献中关于新intima形成细胞被招募的来源一直存在争议。
通过在大鼠腹主动脉中使用CM-Dil染料的细胞示踪剂注射（见 材料表），我们旨在分析起源于母动脉的内皮细胞在两个不同的FU时间点（第7天和第21天）的新intima形成中的作用（图1）。该模型的一个优点是在动脉瘤缝合之前在亲体动脉 中 直接在体内孵育局部细胞示踪剂，从而在以后的时间点允许FU。 体内 注射技术，如细胞示踪剂孵育，尚未在文献中描述。该技术的一个优点是直接，单点，术 中，体内 注射，这使得模型坚固耐用且可重复。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3-0 resorbable suture</td>
			<td>Ethicon Inc., USA</td>
			<td>VCP428G</td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 non-absorbable suture</td>
			<td>B. Braun, Germany</td>
			<td>G0762563</td>
			<td></td>
		</tr>
		<tr>
			<td>6-0 non-absorbable suture</td>
			<td>B. Braun, Germany</td>
			<td>C0766070</td>
			<td></td>
		</tr>
		<tr>
			<td>9-0 non-absorbable suture</td>
			<td>B. Braun, Germany</td>
			<td>G1111140</td>
			<td></td>
		</tr>
		<tr>
			<td>Atipamezol</td>
			<td>Arovet AG, Switzerland</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Bandpass filter blue</td>
			<td>Thorlabs</td>
			<td>FD1B</td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bandpass filter green</td>
			<td>Thorlabs</td>
			<td>FGV9</td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bipolar forceps</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bicycle spotlight</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Board (20 x 10 cm)</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Indivior, Switzerland</td>
			<td>1014197</td>
			<td></td>
		</tr>
		<tr>
			<td>Camera</td>
			<td>Sony NEX-5R, Sony, Tokyo, Japan</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cannula (27-1/2 G)</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Cell count software</td>
			<td>Image-J version 1.52n, U.S. National Institutes of Health, Bethesda, Maryland, USA, https://imagej.nih.gov/ij/</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CellTracker CM-Dil dye</td>
			<td>ThermoFisher SCIENTIFIC, USA</td>
			<td>C7000</td>
			<td></td>
		</tr>
		<tr>
			<td>Coil-Device</td>
			<td>Styker, Kalamazoo, MI, USA</td>
			<td></td>
			<td>2 cm of Target 360 TM Ultra, 2-mm diameter</td>
		</tr>
		<tr>
			<td>Desinfection</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Eye-lubricant</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Fentanyl</td>
			<td>Sintetica, S.A., Switzerland</td>
			<td>98683</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Flumazenil</td>
			<td>Labatec-Pharma, Switerzland</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fluoresceine</td>
			<td>Curatis AG</td>
			<td>5030376</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Fluorescence microscope</td>
			<td>Olympus BX51, Hamburg, Germany; Cell Sens Dimension Imaging software v1.8</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Foil mask</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Glucose (5%)</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Homeothermic Control Unit, Harvard, Edenbridge, England</td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Isotonic sodium chloride solution (0.9%)</td>
			<td>Fresenius KABI</td>
			<td>336769</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td></td>
			<td></td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Longuettes</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Meloxicam</td>
			<td>Boehringer Ingelheim</td>
			<td>P7626406</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Medetomidine</td>
			<td>Virbac, Switzerland</td>
			<td>QN05CM91</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro needle holder</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Midazolam</td>
			<td>Roche, Switzerland</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Monitoring-system</td>
			<td>Starr Life Sciences Corp., 333 Allegheny Ave, Oakmont, PA 15139, United States</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>O2-Face mask</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Operation microscope</td>
			<td>OPMI, Carl Zeiss AG, Oberkochen, Germany</td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Oxygen</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Rectal temperature probe</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Scalpell</td>
			<td>Swann-Morton</td>
			<td>210</td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Small animal shaver</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Smartphone</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Sodium dodecyl sulfate (0.1%)</td>
			<td>Sigma-Aldrich</td>
			<td>11667289001</td>
			<td></td>
		</tr>
		<tr>
			<td>Soft feed</td>
			<td>Emeraid Omnivore</td>
			<td></td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Soft tissue forceps</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Soft tissue spreader</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Stainless steel sponge bowls</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Stent-Device</td>
			<td>Biotroni, B&#252;lach, Switzerland</td>
			<td></td>
			<td>modified magmaris device, AMS with polymer coating, 6-mm length, 2-mm diameter</td>
		</tr>
		<tr>
			<td>Sterile micro swabs</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Straight and curved microforceps</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Straight and curved microscissors</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Straight and curved forceps</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Surgery drape</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Surgical scissors</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Syringes 1 mL, 2 mL, and 5 mL</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Tape</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Vascular clip applicator</td>
			<td>B. Braun, Germany</td>
			<td>FT495T</td>
			<td></td>
		</tr>
		<tr>
			<td>Yasargil titan standard clip (2x)</td>
			<td>B. Braun Medical AG, Aesculap, Switzerland</td>
			<td>FT242T</td>
			<td>temporary</td>
		</tr>
	</tbody>
</table>

using a cell-tracer injection to investigate the origin of neointima-forming cells in a rat saccular side wall model

显微外科夹闭术形成随后的血流屏障进入颅内动脉瘤，而血管内治疗则依赖于新脑和血栓形成。覆盖新内腔层的内皮细胞的来源尚不清楚。因此，本研究的目的是在已经完善的赫尔辛基大鼠显微外科侧壁动脉瘤模型中研究细胞示踪剂注射后新intima形成细胞的起源。
侧壁动脉瘤是通过在雄性Lewis大鼠中将脱细胞化或重要动脉袋首尾并侧缝合到主动脉而形成的。在用动脉瘤缝合线进行动脉切开术之前，在夹紧的主动脉中进行含有CM-Dil染料的细胞示踪剂注射，以标记相邻血管中的内皮细胞并在随访期间跟踪其增殖（FU）。治疗后进行盘绕（n = 16）或支架置入术（n = 15）。在FU（7天或21天）时，所有大鼠都接受荧光血管造影，然后进行动脉瘤收获以及具有特定感兴趣区域的免疫组学细胞计数的宏观和组织学评估。 
31例动脉瘤均未在随访时破裂。四只动物过早死亡。在75.0%盘绕大鼠和7.0%支架大鼠中观察到宏观残余灌注。与盘绕动脉瘤相比，第7天（p = 0.01）和第21天新intima（p = 0.04）的盘绕动脉瘤相比，去细胞化支架的细胞示踪阳性细胞数量显着升高。在血栓或新生血管中未发现显著差异。
这些发现证实，与支架置入式动脉瘤相比，盘绕动脉瘤的愈合模式更差。在去细胞化动脉瘤中，新内膜的形成似乎特别依赖于母动脉，而它是由重要细胞丰富的壁中的动脉瘤壁细胞募集支持的。在转化方面，支架治疗可能更适合于高度退化的动脉瘤，而对于血管壁大多健康的动脉瘤，仅进行盘绕可能就足够了。

实验室环境共纳入31只动物：最终统计分析中包括27只大鼠;4只大鼠过早死亡（死亡率为12.9%）。术中，与线圈处理（13.5μm±0.6）大鼠相比，支架（12.9μm±0.7）的呼吸膨胀显着减少（p = 0.03）。在最后的FU结束时对每只大鼠进行荧光血管造影。在所有6种线圈处理的动物中均有再灌注，而在8种支架处理的动物中仅观察到12.5%的再灌注。
盘管或支架治疗组之间第7天和第21天的?...

Watch this Scientific Journal Video about Using a Cell-Tracer Injection to Investigate the Origin of Neointima-Forming Cells in a Rat Saccular Side Wall Model at JoVE.com

Using a Cell-Tracer Injection to Investigate the Origin of Neointima-Forming Cells in a Rat Saccular Side Wall Model

我们进行了一点亲脂性细胞示踪剂注射以追踪内皮细胞，然后进行动脉切开术和缝合腹大鼠主动脉上的侧壁动脉瘤。在去细胞化动脉瘤中，新intima的形成似乎依赖于母动脉，并且通过从富含细胞的重要壁的动脉瘤壁细胞中募集来促进。

使用细胞示踪剂注射研究大鼠囊侧壁模型中新内膜形成细胞的起源

Microsurgical clipping creates a subsequent barrier of blood flow into intracranial aneurysms, whereas endovascular treatment relies on neointima and ...

通过使用直接的主动脉内细胞示踪剂应用，我们可以分析来自母动脉的内皮细胞在两个不同时间点的新intima形成中的作用。该技术的一大优点是直接，单点，解释性体内注射，这使得该模型成为一种强大且可重复的技术。手术前，将供体大鼠的动脉袋在0.1%十二烷基硫酸钠中在37摄氏度下孵育10小时，以获得去细胞化动脉瘤。在手术前几天从捐赠动物那里收集这些袋子。首先，分析动物的行为并检查粘膜和膨胀，作为术前临床检查的一部分。记录每只动物的体重。对于麻醉诱导，将所有大鼠放在提供氧气的干净盒子中，直到5至10分钟后失去意识。通过没有踏板退出反射来检查麻醉深度。将大鼠置于仰卧位，并用电动剃须刀剃除胸腹区域。使用胶带将老鼠的爪子固定在由连接到自动调节直肠探头的加热垫覆盖的板上。将直肠探针插入大鼠的肛门，在加热垫的帮助下保持37摄氏度的所需温度。然后在连接到计算机系统的右后腿上安装传感器，用于检查术中的生命体征。对于肛周美容护理，在眼睛上涂抹无菌眼科润滑剂，并用不透明的铝箔面罩覆盖它们，以防止手术灯干燥和损坏。用聚维酮碘消毒手术场，并以无菌方式覆盖手术场。在主动脉的近端和远端部分下放置带有紫色衬垫的微拭子，以便在动脉与腔静脉分离后立即更好地可视化动脉。然后用白色纱布保护腹部。在手术当天，通过移液于一毫升PBS溶解两微升细胞示踪剂。将混合物转移到装有套管的一毫升注射器中。关闭手术室的灯。使用显微镜，用微镊子在主动脉的中腹侧部分进行一点注射，并小心地注射一毫升肝素化的0.9%盐水溶液。小心地注射细胞示踪剂，并立即关闭手术显微镜。再次用湿纱布保护腹部。让染料孵育至少15分钟。然后打开显微镜和手术室的灯。使用微镊子和微剪刀进行纵向动脉切开术，使其长度平均为收获的动脉瘤的直径。在进行动脉切开术之前，将动脉瘤放在主动脉旁边，以确保正确的长度。使用不可吸收的10-0缝线，用8至10针缝合动脉瘤。在最后一针之前，将线圈的堆积密度为一厘米。线圈或支架治疗都可以在此模型中进行。然后小心地取下临时夹子，从远端开始，用肝素盐水连续冲洗。以分层方式闭合伤口。在手术结束时，用皮下注射逆转麻醉。让每只操作的动物在干净的笼子里恢复，直到完全清醒，并根据需要用加热灯加热。对于盘管或支架治疗组之间的脱细胞化或重要动脉瘤，第7天和第21天的基线动脉瘤体积没有显着差异。与支架置入式动脉瘤相比，去细胞化动脉瘤的拉动FU体积显示盘绕的动脉瘤生长不显着。拉动FU体积在生命盘绕组中明显大于支架组。在第7天FU时，支架或盘绕处理组新生血管中细胞示踪阳性细胞的数量在支架或盘绕处理组之间没有显着差异，但在第21天时支架大鼠显着较高 FU.No 在7天或21天时在生命动脉瘤缝合大鼠中观察到显着差异，FU.In 7天FU时去细胞化动脉瘤， 与线圈处理的组相比，支架处理的血栓中残留的细胞示踪阳性细胞明显更多。在 FU 七天时，在生命动脉瘤中未观察到这种差异。请记住，在注射细胞示踪剂之前，始终关闭手术室的灯，以防止褪色。另外，让染料孵育至少15分钟。

Research

JoVE Journal

Neuroscience

Gene Delivery to Postnatal Rat Brain by Non-ventricular Plasmid Injection and Electroporation

基因传递到产后大鼠脑非心室质粒注射和电穿孔

Creation of transgenic animals is a standard approach in studying functions of a gene of interest in vivo. However, many knockout or transgenic animals ...

科研

神经科学

Autologous Blood Injection to Model Spontaneous Intracerebral Hemorrhage in Mice

自体血液的注入模型小鼠自发性脑出血

Investigation of the pathophysiology of injury after intracerebral
hemorrhage (ICH) requires a reproducible animal model.  While ICH
accounts for 10-15% ...

Using a Comparative Species Approach to Investigate the Neurobiology of Paternal Responses

使用方法比较物种调查父亲的反应神经生物学

A goal of behavioral neuroscience is to identify underlying neurobiological factors that regulate specific behaviors.  Using animal models to accomplish ...

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

谷氨酸和GABA快速微离子导入：一个有用​​的工具，调查突触整合

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of ...

A Method of Nodose Ganglia Injection in Sprague-Dawley Rat

结状神经节注射液只SD大鼠的方法

Afferent signaling via the vagus nerve transmits important general visceral information to the central nervous system from many diverse receptors located ...

The Use of Trace Eyeblink Classical Conditioning to Assess Hippocampal Dysfunction in a Rat Model of Fetal Alcohol Spectrum Disorders

使用的跟踪经典条件反射，评估胎儿酒精谱系障碍大鼠模型海马功能障碍

Neonatal rats were administered a relatively high concentration of ethyl alcohol (11.9% v/v) during postnatal days 4-9, a time when the fetal brain ...

Biodegradable Magnesium Stent Treatment of Saccular Aneurysms in a Rat Model - Introduction of the Surgical Technique

The steady progess in the armamentarium of techniques available for endovascular treatment of intracranial aneurysms requires affordable and reproducable ...

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method

基于改进多平台法的大鼠中心疲劳模型

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed ...

Direct Injection of a Lentiviral Vector Highlights Multiple Motor Pathways in the Rat Spinal Cord

直接注射慢病毒载体突出大鼠脊髓中的多种运动途径

Introducing proteins of interest into cells in the nervous system is challenging due to innate biological barriers that limit access to most molecules. ...

Facial Nerve Surgery in the Rat Model to Study Axonal Inhibition and Regeneration

大鼠模型中的面部神经外科研究同氧抑制与再生

This protocol describes consistent and reproducible methods to study axonal regeneration and inhibition in a rat facial nerve injury model. The facial ...