Name: extraction of extracellular vesicles from whole tissue
Uploaded: 2019-02-07T14:30:00
Description: Watch this Scientific Journal Video about Extraction of Extracellular Vesicles from Whole Tissue at JoVE.com

作者感谢理查德·诺瓦科夫斯基博士和佛罗里达州立大学实验动物资源组织恭敬地提供和照顾用于开发该协议的动物。我们感谢刘霞、拉凯什·辛格博士和佛罗里达州立大学转化科学实验室在用于下游囊泡表征的质谱研究方面提供的协助, 并感谢 fsu 生物科学成像资源设施。透射电子显微镜在本研究中的应用。最后, 我们感谢 mandip sachdeva 博士 (佛罗里达 a & m 大学) 捐赠了用于开发这种方法的肿瘤标本。这项研究得到了佛罗里达州卫生部 ed 和 ethel moore 阿尔茨海默氏症研究项目的资助, 该项目颁发给了 d. g. m. 和 j. m. (6az11), 并得到了国家卫生研究院国家癌症研究所颁发的奖项。授予 d. g. m. r01 ca204621。

整个大脑都是在佛罗里达州立大学动物使用和护理机构委员会 (iacuc) 的批准下获得的。如前面所述, c57bl6 j 背景中共有12个老鼠大脑 (每个年龄组3个大脑: 2、4、6和 8个月) 用于电动车提取.经佛罗里达农业机械大学 iacuc 批准, mandip sachdeva 博士慷慨捐赠了肺肿瘤标本。肺肿瘤来源于在免疫缺陷的巴布内裸鼠中生长的人腺癌细胞系 h1975。本研究重点介绍了两个具有代表性的肿瘤复制的?...

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对于小电动车在肿瘤微环境中的作用, 以及在器官发育、成熟和功能中的作用, 已经产生了很多科学兴趣。总体而言, 这项研究为从整个大脑或肿瘤标本中提取完整的 ev 提供了优化的工作流程。虽然在这里我们简单地证明了这种技术在肺肿瘤衍生的 ev 中的适用性, 但这种方法可以很容易地适应其他固体组织的工作, 为进一步对小分泌的囊泡进行体内表征提供了机会。广泛的生物医学兴趣。虽然超出?...

小的细胞外囊泡 (ev) 包括内皮衍生的外周细胞和小膜脱落的微囊泡, 具有广泛的生物医学兴趣。小 ev 是由异质种群 50-250 nm 膜结合囊泡, 其中含有生物活性蛋白、脂质和核酸, 这些物质被共同认为在多种疾病过程中发挥着重要作用。推进的研究特别将这些囊泡与神经退行性疾病和 prion 疾病、感染过程、自身免疫性或炎症性疾病以及肿瘤生长和转移1, 2,3有关,4 个,5,6,7.,8,9,10,11,12,13. 对 ev 在疾病发病机制中的重要性的生物医学研究迅速增加, 引起了人们对开发可重复和严格的分离和纯化这些囊泡的方法的平行兴趣。
电动汽车特性的历史和当前挑战是无法完全分离小型 ev 亚群。这一挑战主要是由于我们对不同囊泡的生物发生的不同分子机制的理解有限。亚群之间的重叠大小、密度和生物货物进一步控制了这些区别。这一挑战的部分原因还包括使用截然不同的浓缩技术, 在对实验室间孤立的囊泡进行下游分析方面存在不一致之处, 并破坏了全球为照亮分类电动车种群所做的努力。
值得注意的是, 大多数 ev 特性都是通过体外收集细胞培养上清液进行的, 最近的体内研究描述了从包括血浆、尿液在内的动物或人体体液中分离囊泡的技术, 和唾液。虽然在循环中大量存在的电动车, 但它也认识到, 这些囊泡在细胞间的通信事件中起着重要作用, 并存在于细胞组织的间质中。在癌症的背景下, 间质 ev 可能是特别重要的调节肿瘤微环境的癌细胞种子和转移生长 14,15。因此, 从固体组织标本中提取囊泡的技术的发展和优化具有重要价值。这些方法将提供一种方法, 直接研究有机或肿瘤衍生的 ev 从临床标本, 包括小活检和部分或全部器官切除。
在这项研究中, 以及在我们实验室16发表的上一份报告中, 我们的目标是解决目前在 ev 浓缩方法中关注的几个主要问题: 1) 描述一种可重复的技术, 以分离和净化 ev, 达到目前的最高标准。在该领域被接受;2) 试图分离高富集于内骨源性外显体的小型 ev 亚群;3) 提供从固体组织标本中提取这些囊泡的协议, 以便进一步表征。
最近, kwal 和他的同事描述了一个相对小规模的碘沙诺密度梯度, 以更高的效率分离和纯化 ev 亚群, 比可比的蔗糖密度梯度17。在所引用的研究中, 树突状细胞衍生的 ev 在相对较轻的密度分数下捕获, 与 1.1 g/ml 的密度一致, 在被认为与本报告中存在的高比例外显体的内染色体蛋白中得到了高度丰富的。分数。根据作者的研究, 这些 "真正的" 体外蛋白包括肿瘤易感基因 101 (tsg101)、合成蛋白1、cd81、adam10、ehd4 和几种环素蛋白 17。我们后来调整了这一技术, 以成功的组织离解方法描述的佩雷斯-冈萨雷斯等人18和随后的差分离心协议, 以分离整个脑源 evs16。我们还证明了这种方法在描述 ev 蛋白的效用, 结合顺序协议的下游定量和比较质谱的水泡蛋白, 以前在我们的实验室 19描述。这项工作与希尔实验室的工作是一致的, 在那里, ev 从大脑的额叶皮层丰富了 20。
在这项研究中, 我们详细阐述了这一技术, 并扩大了最近从我们的实验室公布的协议的应用, 以分离电动车从固体肺肿瘤。据我们所知, 这是首次描述从体内肿瘤标本中丰富 evs 的方案的研究。鉴于人们对电动车作为新的诊断生物标志物的广泛兴趣及其在肿瘤形成中的作用, 这种方法可能会对越来越多的科研人员有价值。从临床角度来看, 间质 ev 具有很大的诊断价值, 特别是在组织学评价有限的标本中。我们希望这里概述的方法将为从新鲜或冷冻动物或人类手术标本中获取 ev 的可重复技术提供基础, 为今后的工作铺平道路, 以揭示这些小疾病发病机制中的重要作用小泡可能会发挥。

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">0.45 &#181;m filter</td>
			<td style="width:183px;">VWR</td>
			<td style="width:176px;">28145-505</td>
			<td style="width:355px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">12 mL ultracentrifuge tubes</td>
			<td style="width:183px;">Beckman Coulter</td>
			<td style="width:176px;">331372</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">5.5 mL ultracentrifuge tubes</td>
			<td style="width:183px;">Beckman Coulter</td>
			<td style="width:176px;">344057</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">anti-Alix antibody</td>
			<td style="width:183px;">Santa Cruz</td>
			<td style="width:176px;">sc-7129</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">anti-Calnexin antibody</td>
			<td style="width:183px;">Santa Cruz</td>
			<td style="width:176px;">sc-11397</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">anti-CD63 antibody</td>
			<td style="width:183px;">Abcam</td>
			<td style="width:176px;">ab59479</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">anti-CD81 antibody</td>
			<td style="width:183px;">Santa Cruz</td>
			<td style="width:176px;">sc-9158</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">anti-Flotillin 2 antibody</td>
			<td style="width:183px;">Santa Cruz</td>
			<td style="width:176px;">sc-25507</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">anti-HSC70 antibody</td>
			<td style="width:183px;">Santa Cruz</td>
			<td style="width:176px;">sc-7298</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">anti-Syntenin-1 antibody</td>
			<td style="width:183px;">Santa Cruz</td>
			<td style="width:176px;">sc-100336</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">anti-TSG101 antibody</td>
			<td style="width:183px;">Santa Cruz</td>
			<td style="width:176px;">sc-7964</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">Dounce homogenizer</td>
			<td style="width:183px;">DWK Life Sciences</td>
			<td style="width:176px;">885300-0015</td>
			<td>Loose-fit pestle (clearance of 0.889&#8211;0.165&#8239;mm) used.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">EZQ protein quantification kit</td>
			<td style="width:183px;">ThermoFisher Scientific</td>
			<td style="width:176px;">R33200</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">FA-45-6-30 rotor&#160;</td>
			<td style="width:183px;">Eppendorf</td>
			<td style="width:176px;">5820715006</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">FEI CM120 Electron Microscope</td>
			<td style="width:183px;">TSS Microscopy</td>
			<td style="width:176px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">goat anti-rabbit IgG (Fab fragment)</td>
			<td style="width:183px;">Genetex</td>
			<td style="width:176px;">27171</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">HALT phosphatase inhibitor (100x solution)</td>
			<td style="width:183px;">ThermoFisher Scientific</td>
			<td style="width:176px;">78420</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">HALT protease inhibitor (100x solution)</td>
			<td style="width:183px;">ThermoFisher Scientific</td>
			<td style="width:176px;">78438</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">Hibernate E medium</td>
			<td style="width:183px;">ThermoFisher Scientific</td>
			<td style="width:176px;">A1247601</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">MLS-50 swinging-bucket rotor</td>
			<td style="width:183px;">Beckman Coulter</td>
			<td style="width:176px;">367280</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">NanoSight LM10</td>
			<td style="width:183px;">Malvern</td>
			<td style="width:176px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Optima MAX-XP Benchtop Ultracentrifuge&#160;</td>
			<td style="width:183px;">Beckman Coulter</td>
			<td style="width:176px;">393315</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">Optima XE-100 ultracentrifuge</td>
			<td style="width:183px;">Beckman Coulter</td>
			<td style="width:176px;">A94516</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">Optiprep</td>
			<td style="width:183px;">Sigma</td>
			<td style="width:176px;">D1556</td>
			<td>60% iodixanol in sterile water solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">Q Exactive HF Mass Spectrometer</td>
			<td style="width:183px;">ThermoFisher Scientific</td>
			<td style="width:176px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">rabbit anti-goat IgG</td>
			<td style="width:183px;">Genetex</td>
			<td style="width:176px;">26741</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">rabbit anti-mouse IgG</td>
			<td style="width:183px;">Genetex</td>
			<td style="width:176px;">26728</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">Refracto 30PX (refractometer)</td>
			<td style="width:183px;">Mettler Toledo</td>
			<td style="width:176px;">51324650</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">S-4-104 rotor</td>
			<td style="width:183px;">Eppendorf</td>
			<td style="width:176px;">5820759003</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">SW 41 Ti swinging-bucket Rotor</td>
			<td style="width:183px;">Beckman Coulter</td>
			<td style="width:176px;">333790</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:304px;">Tabletop 5804R centrifuge</td>
			<td style="width:183px;">Eppendorf</td>
			<td style="width:176px;">22623508</td>
			<td></td>
		</tr>
	</tbody>
</table>

extraction of extracellular vesicles from whole tissue

循环和间质小膜结合囊泡 (ev) 是开发新的诊断或预后生物标志物检测的有希望的目标, 并可能成为在大量的进展的重要参与者疾病。目前的研究重点是从多种细胞和组织类型分泌的囊泡的特征, 以便更好地了解 ev 在神经退行性变、炎症和癌症等疾病的发病机制中的作用。然而, 全球一致和可重复的分离和纯化囊泡的技术仍在进行中。此外,从体内固体组织中提取 ev 的方法也很少被描述。在这里, 我们提供了一个详细的协议, 从整个新鲜或冷冻组织, 包括大脑和肿瘤标本中提取感兴趣的小电动车, 以便进一步表征。我们证明了该方法在多个下游分析中的适应性, 包括电子显微镜和囊泡的免疫表型表征, 以及 ev 蛋白的定量质谱。

图 1显示了囊泡的组织离解、差动离心和梯度纯化的示意图。图 2突出了梯度纯化 ev 的形态和免疫表型确认。显示了10-30 碘沙诺梯度超离心后的可重复密度图, 两个不同的囊泡种群向上迁移到分数 2 (轻 ev) 和分数 5 (密集 ev), 取决于组织类型。梯度分数的代表性免疫细胞表现出有效的分离和纯化小肿瘤衍生的 ev 在分数5。值得注意...

Watch this Scientific Journal Video about Extraction of Extracellular Vesicles from Whole Tissue at JoVE.com

Extraction of Extracellular Vesicles from Whole Tissue

全组织细胞外囊的提取

在这里, 我们提供了一个详细的方案, 以分离细胞外囊泡 (ev) 从整个组织, 包括大脑和肿瘤标本。该方法提供了一种可重复的技术, 可从固体组织中提取 ev, 以便进一步进行下游分析。

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

该协议为从整个组织标本中提取和分离小细胞外囊泡提供严格且可重复的技术，用于下游外活分析。在目前可实现的纯度最高水平之一，这种方法有助于直接形态学，免疫性，和深表分泌到各种组织标本，包括肿瘤。在这里，我们演示了从脑和肺肿瘤标本中分离组织 EV。然而，这项技术也可以应用于使用多样化、良性或其他肿瘤标本的研究。首先，在冬眠-E培养基中为每0.4至1.0克组织准备10毫升分离缓冲液。在50毫升管中将整个新鲜或冷冻组织加入缓冲液中，在37摄氏度的温水浴中孵育20分钟。在此之后，添加蛋白酶和磷酸酶抑制剂，在分离缓冲液中进行最终1倍浓度。将溶液与组织倒入松动的软质均质器中。每个样品使用大约 30 次慢冲程轻轻地分离组织。然后，将分离的组织和缓冲液转移到50毫升锥形管中。在500x g和4摄氏度下离心5分钟，使细胞和剩余的纤维或有凝聚力的组织碎片产生颗粒。将上清液转移到一个干净的50毫升锥形管中，在2000xg和4摄氏度下离心10分钟，以颗粒和丢弃大型细胞碎片。将这种上清液转移到一个干净的50毫升圆锥管中，在10000xg和4摄氏度下离心40分钟，以产生任何不需要的较大囊泡或小凋亡体。通过0.45微米过滤器将上清液放入干净的12毫升超离心管中。接下来，超离离样品在10万xg和4摄氏度两个小时，以颗粒小电动汽车。除上经剂，使超离心管倒置 5 至 10 分钟，频繁敲击以去除管两侧的任何残留液体。然后，将 EV 颗粒重新在 1.5 毫升 0.25 摩尔蔗糖缓冲液中重新填充。用 Parafilm 盖住管子，然后将 EV 旋涡进入溶液中。在室温下摇动超离心管 10 至 15 分钟。然后再次漩涡。以低于 1，000x g 的速度短暂离车管，以回收管底的液体悬浮液。如果需要，请将悬挂温度在四摄氏度。首先，在含有 EV 的蔗糖三叶醇缓冲液的 1.5 毫升中加入 1.5 毫升 60%碘糖醇，以创建含有 30%碘糖醇的最终溶液。上下移液多次，将溶液彻底混合。将溶液转移到 5.5 毫升超离心管的底部。接下来，将 60%碘糖醇库存与超纯水混合，以准备至少 1.5 毫升的 20% 和 10% 碘沙醇溶液。使用注射器和 18 量表针，测量 20%碘糖醇溶液的 1.3 毫升，并小心地将它分层在底部梯度的顶部。保持针头的斜面与管内接触，就在半月板的上方，并添加溶液滴明智，以避免在密度界面混合层。然后，使用同样的技术，在20%层顶部对10%碘糖醇溶液的1.2毫升进行分层。小心地平衡超离心管，将输转管装入转子铲斗中。将回转桶转子的加速和减速速度设定为最低速率，并在 268，000x g 和 4 摄氏度下离心 50 分钟。当样品离心时，为每个样品贴上10个1.5毫升微离心管的标签，这些微离心管与密度梯度的一到10的分数相对应。离心完成后，轻轻地从转子铲斗中取出管子，并将其放入稳定的支架中。移液 10 串行分数 490 微升从梯度的顶部到相应的管。使用折射计测量分数的折射率。然后，将每个分数转移到一个干净的12毫升超离心管。在每个管中加入5毫升1x PBS，然后缓慢地上下移液器混合。在管顶部再加入六毫升 1x PBS，然后小心再混合。超离心管在10万xg和4摄氏度，以重新颗粒的小囊泡。去清液，然后轻敲管子，然后将囊泡拉干，进行蛋白质分析或重新切除子宫内进行形态分析。为了对EV进行蛋白质分析，在EV颗粒中加入40微升含有蛋白酶抑制剂的强解解缓冲液。将 Parafilm 放在每个管子上，并大力旋转。接下来，在室温下摇动管子20分钟，并再次旋转。将样品以1000xg快速离心30至60秒，以恢复整个样品体积。将每个样品转移到新的 1.5 毫升微离心管中，并将其储存在 20 至 80 摄氏度的温度下，直到准备进一步处理。要准备纯化的乳酸盐进行免疫布洛特分析，在样品中加入5倍Laemmli样品缓冲液，最终浓度为1倍。将样品在95摄氏度下煮5至10分钟。然后，将一到 10 的等量分数加载到 10% SDS 页面凝胶中。加载等量的组织同质化。执行电泳和西印迹分析，确认纯化的解毒物中的EV蛋白，并比较分数中的相对EV丰度。在这项研究中，从整个组织中提取和纯化细胞外囊泡。在10至30%碘沙醇梯度的超离心化后，可以看到光 EV 的迁移量高达第二位，而根据组织类型，可以看到密集 EV 的迁移量迁移至第 5 部分。梯度分数的代表性免疫扩散在分数五中表现出小肿瘤衍生 EV 的高效分离和纯化。值得注意的是，与以前从整个脑组织采集的光 EV 相比，肺肿瘤标本在密集的 EV 中显得丰富。具有代表性的纳米粒子跟踪分析和电子显微镜组织衍生囊泡在主要囊泡包含分数证明整个囊泡的浓缩和保存。与已知尺寸和结构一致的小型 EV。间向性电动汽车是开发新型诊断或预后生物标志物测定的有希望的目标。该技术为研究人员提供了提取和纯化囊泡的工具，用于下游实用。提取完整或裂解组织 EV 后，可对囊泡进行广泛表征，包括蛋白质组、基因组和唇部分析。此外，示例可用于更有针对性的方法。直接从组织标本中分离的囊泡可以进一步洞察疾病机制，包括肿瘤发生，并可能在未来提供重要的诊断或预测工具。

Research

JoVE Journal

Cancer Research

Neutrophil Extracellular Traps Generated by Low Density Neutrophils Obtained from Peritoneal Lavage Fluid Mediate Tumor Cell Growth and Attachment

腹腔灌洗液中低密度中性粒细胞分泌的中性粒细胞胞外陷阱介导肿瘤细胞生长与附着

Activated neutrophils release neutrophil extracellular traps (NETs), which can capture and destroy microbes. Recent studies suggest that NETs are involved ...

科研

癌症研究

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 ...

Extraction of Aqueous Metabolites from Cultured Adherent Cells for Metabolomic Analysis by Capillary Electrophoresis-Mass Spectrometry

毛细管电泳质谱法提取培养细胞中的水代谢物进行代谢组学分析

Metabolomic analysis is a promising omics approach to not only understand the specific metabolic regulation in cancer cells compared to normal cells but ...

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 ...

Isolation and Functional Assessment of Human Breast Cancer Stem Cells from Cell and Tissue Samples

从细胞和组织样本中分离和功能评估人类乳腺癌干细胞

Breast cancer stem cells (BCSCs) are cancer cells with inherited or acquired stem cell-like characteristics. Despite their low frequency, they are 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 ...

Generating and Imaging Mouse and Human Epithelial Organoids from Normal and Tumor Mammary Tissue Without Passaging

从正常和肿瘤乳腺组织中生成和成像小鼠和人上皮类器官，无需传代

Organoids are a reliable method for modeling organ tissue due to their self-organizing properties and retention of function and architecture after ...

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

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 ...

Pancreatic Cancer-Derived Tumor Organoids and Fibroblasts from Fresh Tissue

Establishment of Pancreatic Cancer-Derived Tumor Organoids and Fibroblasts From Fresh Tissue

Establishment of Pancreatic Cancer-Derived Tumor Organoids and Fibroblasts From Fresh Tissue (Video) | JoVE 

从新鲜组织中建立胰腺癌来源的肿瘤类器官和成纤维细胞

Tumor organoids are three-dimensional (3D) ex vivo tumor models that recapitulate the biological key features of the original primary tumor tissues. ...

Diagnosis of Vitreoretinal Lymphoma Using Cell-Free DNA

Cell-Free DNA Extraction of Vitreous and Aqueous Humor Specimens for Diagnosis and Monitoring of Vitreoretinal Lymphoma

Advancing VRL Diagnosis Using Cell-Free DNA Extraction from Vitreous Humor (Video) | JoVE

玻璃体和房水标本的游离 DNA 提取用于玻璃体视网膜淋巴瘤的诊断和监测

Vitreoretinal lymphoma (VRL) represents an aggressive lymphoma, often categorized as primary central nervous system diffuse large B-cell lymphoma. To ...