Name: ultrasound-guided intracardiac injection of human mesenchymal stem cells to increase homing to the intestine for use in murine models of experimental inflammatory bowel diseases
Uploaded: 2017-09-01T12:30:00
Description: Watch this Scientific Journal Video about Ultrasound-guided Intracardiac Injection of Human Mesenchymal Stem Cells to Increase Homing to the Intestine for Use in Murine Models of Experimental Inflamma

马尼斯戴夫是由国防部补助金 PR141774 和克罗恩和结肠炎基金会美国职业发展奖的支持。Cominelli 的实验室由 NIH 赠款 DK042191 (俱乐部)、DK055812 (俱乐部)、DK091222 (俱乐部) 和 DK07948 (俱乐部) 支持。阿诺德卡普兰的实验室部分由 l. 大卫和 e. 弗吉尼亚鲍德温基金会支持。供资机构在研究分析或手稿的撰写方面没有任何作用。它的内容完全是作者的责任。

该研究中的所有小鼠实验和程序均通过案例西储大学和 #39 的机构动物保育和使用委员会批准。这些程序是在实验室动物保育 (AAALAC) 认可设施的评估和认可协会上进行的。BM 是根据大学医院机构审查委员会批准的协议在干细胞核心设施的情况下, 西储备大学在知情同意后. 
 1. 人骨髓间充质干细胞的培养和转导 (hMSCs) 注意: 在以前的出版物中详细介绍了 hMSC...

<ol>
	<li>Duijvestein, 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=Pretreatment+with+interferon-gamma+enhances+the+therapeutic+activity+of+mesenchymal+stromal+cells+in+animal+models+of+colitis.">Pretreatment with interferon-gamma enhances the therapeutic activity of mesenchymal stromal cells in animal models of colitis.</a> Stem Cells. 29 (10), 1549-1558 (2011).</li><li>Sala, 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=Mesenchymal+Stem+Cells+Reduce+Colitis+in+Mice+via+Release+of+TSG6,+Independently+of+Their+Localization+to+the+Intestine.">Mesenchymal Stem Cells Reduce Colitis in Mice via Release of TSG6, Independently of Their Localization to the Intestine.</a> Gastroenterology. 149 (1), 163-176 (2015).</li><li>Gonzalez, M. A., Gonzalez-Rey, E., Rico, L., Buscher, D., Delgado, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adipose-derived+mesenchymal+stem+cells+alleviate+experimental+colitis+by+inhibiting+inflammatory+and+autoimmune+responses.">Adipose-derived mesenchymal stem cells alleviate experimental colitis by inhibiting inflammatory and autoimmune responses.</a> Gastroenterology. 136 (3), 978-989 (2009).</li><li>Dave, 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=Stem+cells+for+murine+interstitial+cells+of+cajal+suppress+cellular+immunity+and+colitis+via+prostaglandin+e2+secretion.">Stem cells for murine interstitial cells of cajal suppress cellular immunity and colitis via prostaglandin e2 secretion.</a> Gastroenterology. 148 (5), 978-990 (2015).</li><li>Manieri, N. 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=Mucosally+transplanted+mesenchymal+stem+cells+stimulate+intestinal+healing+by+promoting+angiogenesis.">Mucosally transplanted mesenchymal stem cells stimulate intestinal healing by promoting angiogenesis.</a> J. Clin. Invest. 125 (9), 3606-3618 (2015).</li><li>Kontoyiannis, D., Pasparakis, M., Pizarro, T. T., Cominelli, F., Kollias, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+on/off+regulation+of+TNF+biosynthesis+in+mice+lacking+TNF+AU-rich+elements:+implications+for+joint+and+gut-associated+immunopathologies.">Impaired on/off regulation of TNF biosynthesis in mice lacking TNF AU-rich elements: implications for joint and gut-associated immunopathologies.</a> Immunity. 10 (3), 387-398 (1999).</li><li>Abraham, C., Cho, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammatory+bowel+disease.">Inflammatory bowel disease.</a> N. Engl. J. Med. 361 (21), 2066-2078 (2009).</li><li>Dave, M., Papadakis, K. A., Faubion, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunology+of+inflammatory+bowel+disease+and+molecular+targets+for+biologics.">Immunology of inflammatory bowel disease and molecular targets for biologics.</a> Gastroenterol. Clin. North Am. 43 (3), 405-424 (2014).</li><li>Dave, 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=Mesenchymal+Stem+Cell+Therapy+for+Inflammatory+Bowel+Disease:+A+Systematic+Review+and+Meta-analysis.">Mesenchymal Stem Cell Therapy for Inflammatory Bowel Disease: A Systematic Review and Meta-analysis.</a> Inflamm. Bowel Dis. 21 (11), 2696-2707 (2015).</li><li>Panes, 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=Expanded+allogeneic+adipose-derived+mesenchymal+stem+cells+(Cx601)+for+complex+perianal+fistulas+in+Crohn's+disease:+a+phase+3+randomised,+double-blind+controlled+trial.">Expanded allogeneic adipose-derived mesenchymal stem cells (Cx601) for complex perianal fistulas in Crohn's disease: a phase 3 randomised, double-blind controlled trial.</a> Lancet. 388 (10051), (2016).</li><li>Lee, R. 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=Intravenous+hMSCs+improve+myocardial+infarction+in+mice+because+cells+embolized+in+lung+are+activated+to+secrete+the+anti-inflammatory+protein+TSG-6.">Intravenous hMSCs improve myocardial infarction in mice because cells embolized in lung are activated to secrete the anti-inflammatory protein TSG-6.</a> Cell Stem Cell. 5 (1), 54-63 (2009).</li><li>Gao, J., Dennis, J. E., Muzic, R. F., Lundberg, M., Caplan, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+dynamic+in+vivo+distribution+of+bone+marrow-derived+mesenchymal+stem+cells+after+infusion.">The dynamic in vivo distribution of bone marrow-derived mesenchymal stem cells after infusion.</a> Cells Tissues Organs. 169 (1), 12-20 (2001).</li><li>English, K. <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+mesenchymal+stromal+cell+immunomodulation.">Mechanisms of mesenchymal stromal cell immunomodulation.</a> Immunol Cell Biol. 91 (1), 19-26 (2013).</li><li>Pizarro, T. T., 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=SAMP1/YitFc+mouse+strain:+a+spontaneous+model+of+Crohn's+disease-like+ileitis.">SAMP1/YitFc mouse strain: a spontaneous model of Crohn's disease-like ileitis.</a> Inflamm. Bowel Dis. 17 (12), 2566-2584 (2011).</li><li>Lennon, D. P., Caplan, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+human+marrow-derived+mesenchymal+stem+cells.">Isolation of human marrow-derived mesenchymal stem cells.</a> Exp. Hematol. 34 (11), 1604-1605 (2006).</li><li>Lin, 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=Serial+transplantation+and+long-term+engraftment+of+intra-arterially+delivered+clonally+derived+mesenchymal+stem+cells+to+injured+bone+marrow.">Serial transplantation and long-term engraftment of intra-arterially delivered clonally derived mesenchymal stem cells to injured bone marrow.</a> Mol. Ther. 22 (1), 160-168 (2014).</li><li>Dennis, J. E., Caplan, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Porous+ceramic+vehicles+for+rat-marrow-derived+(Rattus+norvegicus)+osteogenic+cell+delivery:+effects+of+pre-treatment+with+fibronectin+or+laminin.">Porous ceramic vehicles for rat-marrow-derived (Rattus norvegicus) osteogenic cell delivery: effects of pre-treatment with fibronectin or laminin.</a> J. Oral Implantol. 19 (2), 106-115 (1993).</li><li>Lin, 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=Efficient+lentiviral+transduction+of+human+mesenchymal+stem+cells+that+preserves+proliferation+and+differentiation+capabilities.">Efficient lentiviral transduction of human mesenchymal stem cells that preserves proliferation and differentiation capabilities.</a> Stem Cells Transl. Med. 1 (12), 886-897 (2012).</li><li>Love, 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=Imaging+of+mesenchymal+stem+cell+transplant+by+bioluminescence+and+PET.">Imaging of mesenchymal stem cell transplant by bioluminescence and PET.</a> PET. J. Nucl. Med. 48 (12), 2011-2020 (2007).</li><li>Haynesworth, S. E., Goshima, J., Goldberg, V. M., Caplan, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+cells+with+osteogenic+potential+from+human+marrow.">Characterization of cells with osteogenic potential from human marrow.</a> Bone. 13 (1), 81-88 (1992).</li><li>Ishizaka, 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=Intra-arterial+cell+transplantation+provides+timing-dependent+cell+distribution+and+functional+recovery+after+stroke.">Intra-arterial cell transplantation provides timing-dependent cell distribution and functional recovery after stroke.</a> Stroke. 44 (3), 720-726 (2013).</li><li>Schenk, 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=Monocyte+chemotactic+protein-3+is+a+myocardial+mesenchymal+stem+cell+homing+factor.">Monocyte chemotactic protein-3 is a myocardial mesenchymal stem cell homing factor.</a> Stem Cells. 25 (1), 245-251 (2007).</li><li>Hayashi, Y., 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=Topical+implantation+of+mesenchymal+stem+cells+has+beneficial+effects+on+healing+of+experimental+colitis+in+rats.">Topical implantation of mesenchymal stem cells has beneficial effects on healing of experimental colitis in rats.</a> J. Pharmacol. Exp. Ther. 326 (2), 523-531 (2008).</li><li>Kodani, T., 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=Flexible+colonoscopy+in+mice+to+evaluate+the+severity+of+colitis+and+colorectal+tumors+using+a+validated+endoscopic+scoring+system.">Flexible colonoscopy in mice to evaluate the severity of colitis and colorectal tumors using a validated endoscopic scoring system.</a> J. Vis. Exp. (80), e50843 (2013).</li></ol>

本研究介绍了一种新的超声引导心内注射 hMSCs 小鼠实验 CD 模型的方法。这项技术有很高的生存率和成功, 由于针的轨迹可以调整基于 real-time, 高分辨率图像的鼠标左心室, 提供的超声。传递到左心室的好处是, hMSCs 然后分布 intra-arterially 和旁路静脉循环, 从而避免了肺部细胞的聚集。以前, 颈动脉插管用于动脉分娩, 用于注入 MSCs16,21。颈动脉插管法是指?...

克罗恩病 (CD) 是一种常见的慢性炎症性疾病的小和大的肠道和被认为是由不适当的反应宿主免疫系统的肠道微生物7,8。最近的研究表明, 小鼠和人骨髓间充质干细胞 (msc) 可以抑制炎症的老鼠模型的肠道炎症1,3,4,5。有多个正在进行的临床试验, 使用从骨髓或脂肪组织获得的人骨髓间充质干细胞治疗炎症性肠病 (IBD) 患者, 其中包括 CD9。在这些临床试验中使用了两种 msc 治疗方法: 一种是对腔 IBD (包括 CD) 的骨髓间充质干细胞进行全身输注 (即静脉注射), 另一条涉及在瘘管中应用/注射骨髓细胞。肛周患者的道。在最近一次 meta 分析中, MSC 治疗 IBD, 系统性 (即,静脉) msc 治疗腔 IBD (包括 CD) 是有效的多达 40% (95% ci: 7-79%) 的患者, 而疗效更高, 在 61% (95% ci:36-85%), 当 MSCs局部注射到患病 CD 瘘9。最近的 III 期多中心随机安慰剂对照试验, 直接注入 CD 患者肛周瘘的同种异体脂肪干细胞, 有统计学意义的临床和放射学证据表明肛周瘘愈合,确证 meta 分析的结果10。腔镉的静脉化疗疗效低下的原因未得到充分调查, 但原因之一可能是 MSCs 对炎症部位的归巢不足。在结肠炎症模型中的小鼠研究表明, 只有一小部分的 mscs (1-5%) 静脉注射到发炎的结肠; 其余的 mscs 由肺部过滤 (第一次通过效果)1,2 ,5,11,12。因此, 多项小鼠研究已使用腹腔路径 (ip) 的 MSC 管理的动物模型结肠炎4。然而, 他们也表明, 只有一小部分细胞到达和嫁接的结肠, 并与疗效相关的可溶性分泌因子的分泌, 如肿瘤坏死因子诱导基因6蛋白 (TSG-6)2。免疫抑制和愈合的 MSC 机制涉及下的方法, 涉及分泌;细胞接近度-独立因素, 如 TSG-6;和细胞接近相关因素, 如程序性死亡-配体 1 (PD-L1);或锯齿状1。因此, MSC 局部化到炎症部位可能会导致效果提高9,13。事实上, 最近的一项研究表明, 在结肠损伤部位直接植入的骨髓间充质干细胞通过分泌血管内皮生长因子 (VEGF) 促进了愈合。另一方面, 静脉注射后注意到了最小的愈合效果5。为了增加其对炎症部位的定位 (即,桑普小鼠的小肠), 该超声引导下的心内注射技术在左心室的 MSC 管理中得到了发展。图像引导注射确保了准确的注射, 从而导致更高的成功率和降低发病率和死亡率。此外, 向左心室注射骨髓间充质干细胞可使它们进入动脉循环, 在那里它们可以到达发炎的小肠, 然后被困在肺部。
在本研究中, 人骨髓间充质干细胞 (hMSCs) 用于注射 SAMP-1/YitFc (桑普) 小鼠模型的 CD14。桑普是一种良好自发的小鼠慢性炎症模型, 它能发展出近100% 显14的小肠炎症。在没有任何化学、免疫学或基因操作的情况下, 炎症发生在对微生物群的反应中, 与人类 CD11相似。本研究采用性别和年龄匹配的无炎症 AKR/J (AKR) 小鼠, 即桑普的父母对照小鼠。
hMSCs 在实验室中从正常的、不明的捐赠者获得的骨髓样本中分离和扩展, 使用经过验证和以前发布的协议15,16。在分离和扩展后, 在实验室中通过多项试验对成骨、脂和软骨分化的 MSC 能力进行了评价15。通过在免疫缺陷 CB17-Prkdc 小鼠体内植入含有 hMSCs 皮的羟基磷灰石/磷酸磷酸盐基质的陶瓷立方体, 进行成骨功能测定17。立方体检测显示成骨和软骨电位, 被认为是评估个别 MSC 准备工作的终极测试17。为了形象化 hMSCs在体内注射后, 慢被用来传感器 hMSCs 与三重报告基因结构, 包括萤火虫荧光 (佛罗里达州), 单体红色荧光蛋白 (mRFP), 和单纯疱疹病毒胸苷激酶 (ttk), 由改良的骨髓肉瘤病毒 (国防部) 启动子18驱动。萤火虫荧光在三重记者是一种酶, 羽注入素氧化在 hMSCs 和产生光子/白光。这是检测的敏感电荷耦合器件 (CCD) 相机 (生物荧光) 在一个体内光学成像系统, 使可视化活 hMSCs 在小鼠。生物发光成像 (BLI) 是一种敏感的技术, 可以连续用于跟踪细胞和ex 体内分析。使用一个强大的国防部启动器驱动的三重融合记者基因结构的连续表达, 并允许成像的注入 hMSCs 超过16周19。hMSCs 不易传感器, 且转导效率低。利用优化的协议, 提高了 hMSCs 转导效率, 提高了转基因表达的18。利用 mRFP 表达 (三重报告基因之一) 对流式细胞术, 传感器 hMSCs 的能力, 高效率高达83% 的证明。鉴别分析和在体内立方体分析证明了转 hMSCs 分化成软骨细胞、脂肪和骨17的能力。

<table border="0" cellpadding="0" cellspacing="0" width="1100">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">DMEM-LG&#160;&#160;</td>
			<td style="width:333px;">Gibco</td>
			<td style="width:188px;">31600-091</td>
			<td style="width:285px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.25% trypsin-EDTA</td>
			<td>Gibco</td>
			<td>25-200-072</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Proteamine Sulfate</td>
			<td>Sigma Aldrich</td>
			<td>P4020-1G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">D-Luciferin</td>
			<td>Goldbio</td>
			<td>luck-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fetal Bobine Serum</td>
			<td>Gibco</td>
			<td>26140-079</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Antibiotic-Antimycotic</td>
			<td>Gibco</td>
			<td>15240-062</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PBS</td>
			<td>HyClone</td>
			<td>SH30256.01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">175 cm tissue cutlure flasks</td>
			<td>Corning</td>
			<td>431080</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">75 cm tissue cutlure flasks</td>
			<td>Corning</td>
			<td>430720</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Centrifuge tubes</td>
			<td>Crysalgen</td>
			<td>23-2265</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tissue-Plus O.C.T. compound</td>
			<td>Fisher HealthCare</td>
			<td>4585</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cryomold Standard</td>
			<td>Tissue-Tek&#160;</td>
			<td>4557</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lenti-Pac HIV Expression Packaging System</td>
			<td>GeneCopoeia</td>
			<td>HPK-LvTR-20&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Povidone Ionidine swabs</td>
			<td>Medline</td>
			<td>MDS093901</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Hair removal cream</td>
			<td style="width:333px;">Nair</td>
			<td style="width:188px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Isoflurane&#160;</td>
			<td style="width:333px;">Piramal Helathcare&#160;</td>
			<td style="width:188px;">NDC 66794 013 25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Forceps&#160;</td>
			<td style="width:333px;">Fine Science Tools</td>
			<td style="width:188px;">11200-33</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Dissection scissors (Wagner)</td>
			<td style="width:333px;">Fine Science Tools</td>
			<td style="width:188px;">Wagner 14068-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Puralube vet ointment</td>
			<td style="width:333px;">Puralube</td>
			<td style="width:188px;">NDC 17033-211-38</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Aquasonic 100 ultrasound transmission gel</td>
			<td style="width:333px;">Parker laboratories</td>
			<td style="width:188px;">01 08</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Petri dish</td>
			<td>Fisher Scientific</td>
			<td>FB0875711A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SAMP mice</td>
			<td>Cleveland Digestive Disease Research Core Centre</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">AKR mice</td>
			<td>Cleveland Digestive Disease Research Core Centre</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vevo 770 imaging system</td>
			<td>&#160;Visual Sonics, Toronto, Canada</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;IVIS spectrum series system&#160;</td>
			<td>PerkinElmer, Waltham, MA</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Living image software&#160;</td>
			<td>CaliperLifeSciences, PerkinElmer, Waltham, MA</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triple reporter</td>
			<td colspan="2">Kindly provided by Dr. Zheng- Hong Lee, CWRU, (citation 19)</td>
			<td></td>
		</tr>
	</tbody>
</table>

ultrasound-guided intracardiac injection of human mesenchymal stem cells to increase homing to the intestine for use in murine models of experimental inflammatory bowel diseases

克罗恩病 (CD) 是一种常见的小、大肠慢性炎症性疾病。小鼠和人骨髓间充质干细胞具有免疫抑制剂的潜能, 并已被证明可以抑制肠道炎症模型中的炎症, 尽管其管理途径可限制其归巢和有效性1,3,4,5. 骨髓间充质干细胞在结肠损伤模型中的局部应用在改善结肠炎症方面表现出更大的功效。然而, 在实验性克罗恩病的 SAMP-1/YitFc (桑普) 模型中, 关于提高人骨髓间充质干细胞 (hMSCs) 在小肠中的定位的技术数据尚缺乏。本文介绍了一种新的超声引导心内注射 hMSCs 在桑普小鼠, 一个良好自发性模型慢性肠道炎症的新技术。性别和年龄匹配, 无炎症 AKR/J (AKR) 小鼠被用作对照。为了分析分布和本土化, hMSCs 转了一个慢, 其中有三名记者。三重记者包括萤火虫荧光 (fl), 为生物发光成像;单体红色荧光蛋白 (mrfp), 用于细胞分类;并截断单纯疱疹病毒胸苷激酶 (ttk), 用于正电子发射断层扫描 (PET) 成像。本研究结果表明, 24 小时后的心内管理, hMSCs 定位在小肠桑普小鼠, 而不是无炎症 AKR 小鼠。这本小说, 超声引导 hMSCs 在左心室的桑普小鼠保证了高成功率的细胞分娩, 使小鼠快速恢复的发病率和死亡率最低。这一技术可以是一种有效的方法, 增强局部化的 MSCs 在其他模型的小肠炎症, 如 TNFΔRE6。未来的研究将确定 hMSCs 通过动脉内分娩增加的局限性是否会导致治疗效果的提高。

图 1显示, hMSCs 可以与三重报告员转, 以高效率地保存其干细胞特性。转 hMSCs 可以通过 BLI (图 1C,图 3) 实时可视化。这本小说, 超声引导注射 hMSCs 进入左心室的桑普小鼠确保了非常高的成功率注射, 允许快速恢复小鼠的发病率和死亡率最低 (图 2)。注射一只小鼠与 hMSCs 的平均时间约10分...

Watch this Scientific Journal Video about Ultrasound-guided Intracardiac Injection of Human Mesenchymal Stem Cells to Increase Homing to the Intestine for Use in Murine Models of Experimental Inflamma

Ultrasound-guided Intracardiac Injection of Human Mesenchymal Stem Cells to Increase Homing to the Intestine for Use in Murine Models of Experimental Inflammatory Bowel Diseases

小鼠在结肠炎症模型中的研究表明, 一小部分 (1-5%) 的间充质干细胞 (MSC) 静脉注射或腹腔回家到发炎的结肠1,2。本研究表明, 超声引导下的骨髓间充质干细胞注射可增加肠道定位。

超声引导下人骨髓间充质干细胞在实验性肠炎小鼠模型中的应用

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have ...

Research

JoVE Journal

Medicine

The Measurement and Treatment of Suppression in Amblyopia

测量和制止在弱视治疗

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

科研

医学

The Use of Primary Human Fibroblasts for Monitoring Mitochondrial Phenotypes in the Field of Parkinson's Disease

利用人类纤维原细胞线粒体表型的监测领域的帕金森氏病

Parkinson's disease (PD) is the second most  common movement disorder and affects 1% of people over the age of 60 1. Because ageing is  the most important ...

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

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 ...

Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

异基因骨髓间充质干细胞植入兔子的膝关节软骨缺损治疗纤维蛋白凝块

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular ...

DNBS/TNBS Colitis Models: Providing Insights Into Inflammatory Bowel Disease and Effects of Dietary Fat

二硝基苯磺酸/ TNBS结肠炎模型：提供洞察炎症性肠病和膳食脂肪的影响

Inflammatory Bowel Diseases (IBD), including Crohn&#39;s Disease and Ulcerative Colitis, have long been associated with a genetic basis, and more recently ...

Ultrasound-guided Transthoracic Intramyocardial Injection in Mice

超声引导下经皮内注射小鼠

Murine models of cardiovascular disease are important for investigating pathophysiological mechanisms and exploring potential regenerative therapies. ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

研究胰腺癌干细胞特性开发新的治疗策略

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Injection of Syngeneic Murine Melanoma Cells to Determine Their Metastatic Potential in the Lungs

同基因小鼠黑色素瘤细胞注射，以确定其在肺转移潜能

Approximately 90% of human cancer deaths are linked to metastasis.  Despite the prevalence and relative harm of metastasis, therapeutics for treatment or ...