Name: an enrichment method for small extracellular vesicles derived from liver cancer tissue
Uploaded: 2023-02-03T12:50:00
Description: Watch this Scientific Journal Video about An Enrichment Method for Small Extracellular Vesicles Derived from Liver Cancer Tissue at JoVE.com

作者感谢赣南医科大学附属第一医院对这项工作的支持。这项工作得到了中国国家自然科学基金（批准号82260422）的支持。

从甘南医科大学附属第一医院诊断为肝恶性肿瘤的患者中采集人肝癌组织。所有患者均签署知情同意书，人体组织样本采集经甘南医科大学第一附属医院伦理委员会批准。有关此协议中使用的所有材料、设备和软件的详细信息，请参阅 材料表 。
1. 准备
<ol><li>将转移脱色摇床放入培养箱中，并将温度设置为37°C。 有关所有其他所需设备，请参见 <stro...

<ol>
	<li>Isaac, R., Reis, F. C. G., Ying, W., Olefsky, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exosomes+as+mediators+of+intercellular+crosstalk+in+metabolism.">Exosomes as mediators of intercellular crosstalk in metabolism.</a> Cell Metabolism. 33 (9), 1744-1762 (2021).</li><li>Hou, 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=Advances+in+exosome+isolation+methods+and+their+applications+in+proteomic+analysis+of+biological+samples.">Advances in exosome isolation methods and their applications in proteomic analysis of biological samples.</a> Analytical and Bioanalytical Chemistry. 411 (21), 5351-5361 (2019).</li><li>Zhang, L., Yu, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exosomes+in+cancer+development,+metastasis,+and+immunity.">Exosomes in cancer development, metastasis, and immunity.</a> Biochimica et Biophysica Acta - Reviews on Cancer. 1871 (2), 455-468 (2019).</li><li>Yang, 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=and+perspective+on+exosome+isolation+-+Efforts+for+efficient+exosome-based+theranostics.">and perspective on exosome isolation - Efforts for efficient exosome-based theranostics.</a> Theranostics. 10 (8), 3684-3707 (2020).</li><li>Crescitelli, R., L&#228;sser, C., L&#246;tvall, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+extracellular+vesicle+subpopulations+from+tissues.">Isolation and characterization of extracellular vesicle subpopulations from tissues.</a> Nature Protocols. 16 (3), 1548-1580 (2021).</li><li>Crescitelli, 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=Subpopulations+of+extracellular+vesicles+from+human+metastatic+melanoma+tissue+identified+by+quantitative+proteomics+after+optimized+isolation.">Subpopulations of extracellular vesicles from human metastatic melanoma tissue identified by quantitative proteomics after optimized isolation.</a> Journal of Extracellular Vesicles. 9 (1), 1722433 (2020).</li><li>Jang, S. 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=Mitochondrial+protein+enriched+extracellular+vesicles+discovered+in+human+melanoma+tissues+can+be+detected+in+patient+plasma.">Mitochondrial protein enriched extracellular vesicles discovered in human melanoma tissues can be detected in patient plasma.</a> Journal of Extracellular Vesicles. 8 (1), 1635420 (2019).</li><li>Muralidharan-Chari, 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=ARF6-regulated+shedding+of+tumor+cell-derived+plasma+membrane+microvesicles.">ARF6-regulated shedding of tumor cell-derived plasma membrane microvesicles.</a> Current Biology. 19 (22), 1875-1885 (2009).</li><li>Matejovi&#269;, A., Wakao, S., Kitada, M., Kushida, Y., Dezawa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+separation+methods+for+tissue-derived+extracellular+vesicles+in+the+liver,+heart,+and+skeletal+muscle.">Comparison of separation methods for tissue-derived extracellular vesicles in the liver, heart, and skeletal muscle.</a> FEBS Open Bio. 11 (2), 482-493 (2021).</li><li>Zhou, 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=Brown+adipose+tissue-derived+exosomes+mitigate+the+metabolic+syndrome+in+high+fat+diet+mice.">Brown adipose tissue-derived exosomes mitigate the metabolic syndrome in high fat diet mice.</a> Theranostics. 10 (18), 8197-8210 (2020).</li><li>Chen, 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=Comparison+of+the+variability+of+small+extracellular+vesicles+derived+from+human+liver+cancer+tissues+and+cultured+from+the+tissue+explants+based+on+a+simple+enrichment+method.">Comparison of the variability of small extracellular vesicles derived from human liver cancer tissues and cultured from the tissue explants based on a simple enrichment method.</a> Stem Cell Reviews and Reports. 18 (3), 1067-1077 (2022).</li><li>Fang, 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=Tumor-derived+exosomal+miR-1247-3p+induces+cancer-associated+fibroblast+activation+to+foster+lung+metastasis+of+liver+cancer.">Tumor-derived exosomal miR-1247-3p induces cancer-associated fibroblast activation to foster lung metastasis of liver cancer.</a> Nature Communications. 9 (1), 1247-1243 (2018).</li><li>Huang, 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=RNA+sequencing+of+plasma+exosomes+revealed+novel+functional+long+noncoding+RNAs+in+hepatocellular+carcinoma.">RNA sequencing of plasma exosomes revealed novel functional long noncoding RNAs in hepatocellular carcinoma.</a> Cancer Science. 111 (9), 3338-3349 (2020).</li><li>Chen, 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=HCC-derived+exosomes+elicit+HCC+progression+and+recurrence+by+epithelial-mesenchymal+transition+through+MAPK/ERK+signalling+pathway.">HCC-derived exosomes elicit HCC progression and recurrence by epithelial-mesenchymal transition through MAPK/ERK signalling pathway.</a> Cell Death &amp; Disease. 9 (5), 513 (2018).</li><li>Jingushi, 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=Extracellular+vesicles+isolated+from+human+renal+cell+carcinoma+tissues+disrupt+vascular+endothelial+cell+morphology+via+azurocidin.">Extracellular vesicles isolated from human renal cell carcinoma tissues disrupt vascular endothelial cell morphology via azurocidin.</a> International Journal of Cancer. 142 (3), 607-617 (2018).</li><li>Camino, 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=Deciphering+adipose+tissue+extracellular+vesicles+protein+cargo+and+its+role+in+obesity.">Deciphering adipose tissue extracellular vesicles protein cargo and its role in obesity.</a> International Journal of Molecular Sciences. 21 (24), 9366 (2020).</li></ol>

该协议描述了从肝癌组织中提取sEV的可重复方法。通过尖锐的组织分离、消化酶处理、差示超速离心和 0.22 μm 滤膜过滤和纯化获得高质量的 sEV。对于下游分析，确保sEV的高纯度非常重要。在差速离心过程中，上清液表面会出现一层白色物质（成分不明）。由于该层会污染sEV，我们通过重复离心（3，000 × g 和12，000 × g）并通过0.22 μm膜过滤（补充表S1）尽可能去除它。由...

小细胞外囊泡（sEV）的直径约为30nm至150nm，由各种细胞分泌1。它们可以与组织细胞交流，通过将重要的生物分子（如脂质、蛋白质、DNA 和 RNA）运送到各种器官、组织、细胞和细胞内部分来调节局部或远处的微环境。因此，它们还可以改变受体细胞2，3的行为。特定sEV的分离和纯化是研究其在疾病发展和过程中的生物学行为的重要先决条件。差速超速离心 - 被视为金标准 - 通常用于将sEV与其通常所在的组织分离4。组织碎片、细胞碎片、大囊泡和凋亡小体可以通过这种技术去除，只留下sEV。
胶原酶D和DNase I已被证明不会影响细胞或囊泡的分子特征，这两种酶的特性都有助于细胞外基质中囊泡的释放5。这些酶已用于从人转移性黑色素瘤组织、结肠癌组织和结肠粘膜组织中提取 sEV5，6，7。然而，这些方法中胶原酶D和DNase I的浓度和消化时间不同，导致结论不一致。为了避免其他sEV亚型的共沉淀，研究人员通过过滤和/或差速离心去除了较大的细胞外囊泡（直径0.1μm或0.2μm8）。根据来源组织的不同，可能需要不同的分离和纯化方法9，10。
使用传统的差示超速离心方法从肝组织中提取sEV会在上清液表面形成一层白质，而无法确定其性质。在之前的研究11中，发现这层白质会影响sEV的纯度。虽然传统方法分离的样品颗粒数和蛋白质浓度高于现有方法，但变异系数较大，可能是因为许多污染物会导致结果的重复性差。也就是说，使用洗涤剂（即检测颗粒在1%Triton X-100中的溶解度），我们发现通过这种方法获得的sEV的纯度更高。因此，我们使用这种方法分离和纯化来自结直肠癌组织的sEV，用于蛋白质组学研究。
目前，sEVs在肝癌中的研究主要集中在血清、血浆和细胞培养物的上清液12，13，14上。然而，来源于肝癌组织的sEV可以更准确地反映肝癌的生理病理和周围微环境，有效避免其他EV的降解和污染15，16。利用差示超速离心，该方法可富产率，获得高质量的sEVs，为肝癌的进一步研究提供重要依据。该方法使肝癌组织能够通过尖锐分离分离，并通过胶原酶D和DNase I解离。然后，通过过滤和差示超速离心进一步去除细胞碎片、大囊泡和凋亡小体。最后，分离并纯化sEV以供后续研究。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.22 &#181;m Membrane Filter Unit</td>
			<td>Millex</td>
			<td>SLGPR33RB</td>
			<td></td>
		</tr>
		<tr>
			<td>1 mL Sterile syringe</td>
			<td>Hubei Xianming Medical Instrument Company</td>
			<td>YL01329</td>
			<td></td>
		</tr>
		<tr>
			<td>2% Uranyl Acetate</td>
			<td>Electron Microscopy Sciences</td>
			<td>22400-2</td>
			<td></td>
		</tr>
		<tr>
			<td>4.7 mL Centrifuge Tube</td>
			<td>Beckman Coulter</td>
			<td>361621</td>
			<td></td>
		</tr>
		<tr>
			<td>6-well Cell cuture plate</td>
			<td>LABSELECT</td>
			<td>11110</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL Beaker</td>
			<td>Tianjin Kangyiheng Experimental Instrument Sales Company</td>
			<td>CF2100800</td>
			<td></td>
		</tr>
		<tr>
			<td>70 &#181;m Cell strainer</td>
			<td>Biosharp</td>
			<td>BS-70-XBS</td>
			<td></td>
		</tr>
		<tr>
			<td>100 mm Cell culture dish</td>
			<td>CELL TER</td>
			<td>CS016-0128</td>
			<td></td>
		</tr>
		<tr>
			<td>600 &#181;L Centrifuge tube</td>
			<td>Axygen</td>
			<td>MCT060C</td>
			<td></td>
		</tr>
		<tr>
			<td>BCA protein quantification kit</td>
			<td>Thermo Fisher</td>
			<td>RJ240544</td>
			<td></td>
		</tr>
		<tr>
			<td>Beckman Coulter Optima-Max-TL</td>
			<td>Beckman&#160;</td>
			<td>A95761</td>
			<td></td>
		</tr>
		<tr>
			<td>BioRad Mini trans-blot</td>
			<td>Bio-Rad</td>
			<td>1703930</td>
			<td></td>
		</tr>
		<tr>
			<td>BioRad Mini-Protean</td>
			<td>Bio-Rad</td>
			<td>1645050</td>
			<td></td>
		</tr>
		<tr>
			<td>CD63 Antibody</td>
			<td>Abcam</td>
			<td>ab134045</td>
			<td></td>
		</tr>
		<tr>
			<td>CD9 Antibody</td>
			<td>Abcam</td>
			<td>ab263019</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge 5430R</td>
			<td>Eppendorf</td>
			<td>5428HQ527333</td>
			<td></td>
		</tr>
		<tr>
			<td>Cleaning Solution</td>
			<td>NanoFCM</td>
			<td>C1801</td>
			<td></td>
		</tr>
		<tr>
			<td>Collagenase D</td>
			<td>Roche</td>
			<td>11088866001</td>
			<td></td>
		</tr>
		<tr>
			<td>Copper net</td>
			<td>Henan Zhongjingkeyi Technology Company</td>
			<td>DJZCM-15-N1</td>
			<td></td>
		</tr>
		<tr>
			<td>Dry Thermostat</td>
			<td>Hangzhou allsheng instruments company</td>
			<td>AS-01030-00</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC Anti Human CD9 Antibody</td>
			<td>Elabscience</td>
			<td>E-AB-F1086C</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycine</td>
			<td>Solarbio</td>
			<td>G8200</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat horseradish peroxidase (HRP)-coupled secondary anti-mouse antibody&#160;</td>
			<td>Proteintech</td>
			<td>SA00001-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat horseradish peroxidase (HRP)-coupled secondary anti-rabbit antibody&#160;</td>
			<td>Proteintech</td>
			<td>SA00001-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Shanghai Zhenxing Chemical Company</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Nanoparticle flow cytometer</td>
			<td>NanoFCM INC</td>
			<td>FNAN30E20112368</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphatase inhibitors(PhosSTOP)</td>
			<td>Roche</td>
			<td>4906845001</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline(PBS)</td>
			<td>Servicebio</td>
			<td>G4202</td>
			<td></td>
		</tr>
		<tr>
			<td>Polyvinylidene Difluoride Membrane</td>
			<td>Solarbio</td>
			<td>ISEQ00010</td>
			<td></td>
		</tr>
		<tr>
			<td>QC Beads</td>
			<td>NanoFCM</td>
			<td>QS2502</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI-1640 basic medium</td>
			<td>Biological Industries&#160;</td>
			<td>C11875500BT</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel</td>
			<td>Guangzhou Kehua Trading Company</td>
			<td>NN-0623-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Silica Nanospheres</td>
			<td>NanoFCM</td>
			<td>S16M-Exo</td>
			<td></td>
		</tr>
		<tr>
			<td>Transference Decoloring Shaker TS-8</td>
			<td>Kylin-Bell</td>
			<td>E0018</td>
			<td></td>
		</tr>
		<tr>
			<td>Transmission Electron Microscope</td>
			<td>Thermo Scientific</td>
			<td>Talos L120C</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris</td>
			<td>Solarbio</td>
			<td>T8060</td>
			<td></td>
		</tr>
		<tr>
			<td>TSG101 Antibody</td>
			<td>Proteintech</td>
			<td>28283-1-AP</td>
			<td></td>
		</tr>
		<tr>
			<td>Tweezer</td>
			<td>Guangzhou Lige Technology Company</td>
			<td>LG01-105-4X</td>
			<td></td>
		</tr>
	</tbody>
</table>

an enrichment method for small extracellular vesicles derived from liver cancer tissue

来源于组织的小细胞外囊泡（sEV）可以反映源细胞的功能状态和组织间质空间的特征。这些sEVs的高效富集是研究其生物学功能的重要前提，也是临床检测技术和治疗载体技术发展的关键。很难从组织中分离出sEV，因为它们通常受到严重污染。本研究提供了一种从肝癌组织中快速富集高质量sEV的方法。该方法涉及一个四步过程：消化酶（胶原酶D和DNase Ι）与组织的孵育，通过70μm细胞过滤器过滤，差分超速离心，并通过0.22μm膜过滤器过滤。由于差示超速离心步骤的优化和过滤步骤的加入，该方法获得的sEV纯度高于经典差示超速离心。它为组织来源的sEV研究提供了重要的方法和支持数据。

来自人肝癌组织的sEV在肝癌患者的诊断、治疗和预后中发挥了至关重要的作用。该方法使用普通的实验室仪器分离和纯化来自肝癌组织的sEV;这可能为sEV的研究提供方法学支持。图2说明了从肝癌组织中富集sEV的一般过程。组织细胞间隙中的sEV通过组织切割和酶水解完全释放。此外，消化溶液通过70μm过滤器过滤以去除大的组织碎片。此外，进行差速离心步骤（500 × g、3...

Watch this Scientific Journal Video about An Enrichment Method for Small Extracellular Vesicles Derived from Liver Cancer Tissue at JoVE.com

An Enrichment Method for Small Extracellular Vesicles Derived from Liver Cancer Tissue

在这里，我们描述了通过优化的差分超速离心方法富集来自肝癌组织的小细胞外囊泡。

肝癌组织来源于小细胞外囊泡的富集方法

Small extracellular vesicles (sEVs) derived from tissue can reflect the functional status of the source cells and the characteristics of the tissue's ...

该方法允许分离肝癌组织来源的小细胞外囊泡，其纯度高于经典的差示超速离心。该技术简单、方便、易于操作，可为小细胞外囊泡的研究提供方法学支持。首先，从 80 摄氏度的冰箱中取出组织样本。将大约400毫克的组织放入冰盒上的10厘米细胞培养皿中，并用手术刀将组织切碎。将组织转移到装有1.5毫升消化液的六孔板中。然后，将板放在移位脱色摇床上，并在37摄氏度下孵育20分钟，以完全解离肝癌组织并释放小细胞外囊泡或sEV。孵育后，将六孔板放在冰盒上。加入80微升磷酸酶抑制剂和200微升完全蛋白酶抑制剂溶液以停止消化。然后，将消化溶液转移到放置在两毫升离心管上的70微米细胞过滤器中，并缓慢过滤以去除任何大的组织碎片。将滤液以500g离心10分钟以除去小组织碎片，并将上清液收集在新的两毫升离心管中。将上清液以3， 000g离心20分钟以除去细胞碎片。小心地将上清液转移到新的离心管中，直到移液器的尖端即将接触底部的白色碎片。重复离心剩余液体后，将上清液汇集到管中而不干扰碎屑。将收集的上清液以12， 000g离心20分钟，并将900微升上清液转移到4.7毫米超速离心管中。重复离心剩余的液体，然后收集并将两种上清液混合到同一超速离心管中。用PBS完全填充管子。将上清液以100， 000g离心60分钟。弃去上清液后，通过用PBS填充超速离心管来重悬沉淀。在以100， 000g重复离心60分钟后，用50微升PBS重悬沉淀。使用一毫升无菌注射器吸出sEV悬浮液，并通过0.22微米膜过滤器将其过滤到600微升离心管中。将滤液储存在80摄氏度下进行进一步分析。使用纳米颗粒流式细胞仪确定sEV的尺寸分布和纯度。检查清洁溶液的体积。然后，注意护套和废液的水平之间的差异。打开仪器主电源20秒后，打开计算机。等待蜂鸣音指示仪器已正确连接以运行软件。单击启动以打开相机、激光器和气泵。接下来，单击护套流，然后选择启动，将超纯水放在装载平台上。四分钟后，单击“采样和提升”，然后选择“在样本中卸载”。30秒后，将空白管放在装载平台上。单击“示例”，选择“提升”，然后选择“在示例中卸载”。然后，将装有超纯水的管放在装载平台上。同时，单击“采样和提升”，然后单击“护套流”和“清除”。单击手动操作，然后将颗粒浓缩管放在装载平台上。然后，单击“样品”，选择“增强”一分钟，同时在“样品信息”中选择“250纳米标准荧光二氧化硅纳米球质量控制”。选择从样本中采样，然后单击 SPCM 打开检测器。然后，单击自动采样，并在采样集中输入 1.0 以将压力固定在一千帕。单击工具栏，然后选择大信号。调整激光器的水平位置，并将激光器设置为两微米。然后，连续单击L和R以确保信号强且均匀。单击“记录时间”以收集数据，完成后会自动跳转到“缓冲区”。然后，将数据保存在 Nfa 文件中，然后单击“示例”以选择“卸载”。将清洁溶液管放在装载平台上。单击“采样”，选择“提升”，一分钟后单击“采样和卸载”。使用超纯水从毛细管尖端去除残留的清洁溶液。将粒径标准管放在装载平台上，然后单击样品提升一分钟。在样品信息中选择 68 到 155 S16M-Exo 质量控制，然后单击样品，然后单击采样。接下来，单击工具栏，然后选择小信号。单击“记录时间”以收集数据，完成后会自动跳转到“缓冲区”。然后，将数据保存在 Nfa 文件中，然后单击“示例”以选择“卸载”。用清洁溶液去除残留物后，将PBS管放在装载平台上，并执行针对粒径标准管演示的步骤。接下来，如前所述清洁毛细管尖端，并加载sEV样品以分析前面描述的粒径标准管的数据。在透射电子显微镜下检查纯化的sEVs，揭示了具有杯形形态的sEV的存在。对sEV进行了流式细胞术。粒径范围从40纳米到200纳米，平均粒径为89纳米。富集sEVs的纯度为68.32%，蛋白质免疫印迹法检测sEV的CD63、CD9、TSG101等蛋白标志物。GM130被用作sEVs的阴性对照标志物，在组织细胞中发现，但在sEV中没有发现。至关重要的是要非常小心地执行酶消化、差示超速离心和膜过滤步骤，以确保小细胞外囊泡的纯度。

Research

JoVE Journal

Cancer Research

Modeling Osteosarcoma Using Li-Fraumeni Syndrome Patient-derived Induced Pluripotent Stem Cells

应用 Fraumeni 综合征诱导多潜能干细胞建模骨肉瘤

Li-Fraumeni syndrome (LFS) is an autosomal dominant hereditary cancer disorder. Patients with LFS are predisposed to a various type of tumors, including ...

科研

癌症研究

Extraction of Extracellular Vesicles from Whole Tissue

全组织细胞外囊的提取

Circulating and interstitial small membrane-bound extracellular vesicles (EVs) represent promising targets for the development of novel diagnostic or ...

Studying Normal Tissue Radiation Effects using Extracellular Matrix Hydrogels

研究使用细胞外基体水凝胶的正常组织辐射效应

Radiation is a therapy for patients with triple negative breast cancer. The effect of radiation on the extracellular matrix (ECM) of healthy breast tissue ...

Spatial and Temporal Control of Murine Melanoma Initiation from Mutant Melanocyte Stem Cells

突变黑色素细胞干细胞引发小鼠黑色素瘤的时空控制

Cutaneous melanoma is well known as the most aggressive skin cancer. Although the risk factors and major genetic alterations continue to be documented ...

Isolation of Exosome-Enriched Extracellular Vesicles Carrying Granulocyte-Macrophage Colony-Stimulating Factor from Embryonic Stem Cells

分离外体富集的细胞外囊泡携带颗粒细胞-巨噬细胞-胚胎干细胞的细胞外刺激因子

Embryonic stem cells (ESCs) are pluripotent stem cells capable of self-renewal and differentiation into all types of embryonic cells. Like many other cell ...

Studying Triple Negative Breast Cancer Using Orthotopic Breast Cancer Model

研究使用正交乳腺癌模型的三阴性乳腺癌

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less ...

Sequencing Small Non-coding RNA from Formalin-fixed Tissues and Serum-derived Exosomes from Castration-resistant Prostate Cancer Patients

从卡西甲抗性前列腺癌患者的甲酸固定组织和血清衍生的外源体对小型非编码RNA进行测序

Ablation of androgen receptor (AR) signaling by androgen deprivation is the goal of the first line of therapy for prostate cancer that initially results ...

Expansion and Enrichment of Gamma-Delta (&#947;&#948;) T Cells from Apheresed Human Product

Expansion and Enrichment of Gamma-Delta ÃƒÅ½Ã‚Â³ÃƒÅ½Ã‚Â´ T Cells from Apheresed Human Product

来自分离人产品的γ-Delta（γδ）T细胞的扩增和富集

Although V&#947;9V&#948;2 T cells are a minor subset of T lymphocytes, this population is sought after for its ability to recognize antigens in a major ...

In Vivo Immunogenicity Screening of Tumor-Derived Extracellular Vesicles by Flow Cytometry of Splenic T Cells

In Vivo Immunogenicity Screening of Tumor-Derived Extracellular Vesicles by Flow Cytometry of Splenic T Cells

体内 通过脾T细胞流式细胞术筛选肿瘤来源的细胞外囊泡的免疫原性

Immunogenic cell death of tumors, caused by chemotherapy or irradiation, can trigger tumor-specific T cell responses by releasing danger-associated ...

A Three-Dimensional Spheroid Model to Investigate the Tumor-Stromal Interaction in Hepatocellular Carcinoma

一种三维球体模型研究肝细胞癌肿瘤-基质相互作用

The aggressiveness and lack of well-tolerated and widely effective treatments for advanced hepatocellular carcinoma (HCC), the predominant form of liver ...