Name: ex vivo organoid model of adenovirus-cre mediated gene deletions in mouse urothelial cells
Uploaded: 2022-05-05T15:00:00
Description: Watch this Scientific Journal Video about Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells at JoVE.com

这项研究工作得到了NIH Grants，K08CA252161（Q.L.），R01CA234162和R01 CA207757（D.W.G.），P30CA016056（NCI癌症中心核心支持补助金），罗斯威尔公园联盟基金会和泌尿科之友基金会的部分支持。我们感谢玛丽莎·布拉斯克和米拉·帕霍莫娃对手稿的校对。

所有动物程序均由纽约州布法罗罗斯威尔公园综合癌症中心的机构动物护理和使用委员会（IACUC）批准（1395M，生物安全180501和180502）。 注意：在同一天执行步骤 1-3。
1. 小鼠膀胱解剖
<ol><li>解剖准备<ol>
<li>准备所有无菌器械，包括剪刀，镊子，35毫米培养皿中的无菌DPBS，70%乙醇，无菌纱布和干净的纸巾。清洁解剖区域的所有表面。生成雄性三重浮选小鼠 Tr...

<ol>
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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=Rb1+and+Trp53+cooperate+to+suppress+prostate+cancer+lineage+plasticity,+metastasis,+and+antiandrogen+resistance.">Rb1 and Trp53 cooperate to suppress prostate cancer lineage plasticity, metastasis, and antiandrogen resistance.</a> Science. 355 (6320), 78-83 (2017).</li><li>Lee, S. 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=Tumor+evolution+and+drug+response+in+patient-derived+organoid+models+of+bladder+cancer.">Tumor evolution and drug response in patient-derived organoid models of bladder cancer.</a> Cell. 173 (2), 515-528 (2018).</li><li>Fujii, 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=A+simple+method+for+histopathological+evaluation+of+organoids.">A simple method for histopathological evaluation of organoids.</a> Journal of Toxicologic Pathology. 31 (1), 81-85 (2018).</li><li>Georgas, K. 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=An+illustrated+anatomical+ontology+of+the+developing+mouse+lower+urogenital+tract.">An illustrated anatomical ontology of the developing mouse lower urogenital tract.</a> Development. 142 (10), 1893-1908 (2015).</li><li>Santos, C. 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=Urothelial+organoids+originating+from+Cd49f(high)+mouse+stem+cells+display+Notch-dependent+differentiation+capacity.">Urothelial organoids originating from Cd49f(high) mouse stem cells display Notch-dependent differentiation capacity.</a> Nature Communication. 10 (1), 4407 (2019).</li><li>Wang, 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=A+genetically+defined+disease+model+reveals+that+urothelial+cells+can+initiate+divergent+bladder+cancer+phenotypes.">A genetically defined disease model reveals that urothelial cells can initiate divergent bladder cancer phenotypes.</a> Proceedings of the National Academy of Sciences of the United States of America. 117 (1), 563-572 (2020).</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=Intertumoral+heterogeneity+in+SCLC+is+influenced+by+the+cell+type+of+origin.">Intertumoral heterogeneity in SCLC is influenced by the cell type of origin.</a> Cancer Discovery. 8 (10), 1316-1331 (2018).</li><li>Sutherland, K. 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K., Jones, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adenovirus-mediated+genetic+removal+of+signaling+molecules+in+cultured+primary+mouse+embryonic+fibroblasts.">Adenovirus-mediated genetic removal of signaling molecules in cultured primary mouse embryonic fibroblasts.</a> Journal of Visualized Experiments: JoVE. (43), e2160 (2010).</li><li>Feng, W., 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=Rapid+interrogation+of+cancer+cell+of+origin+through+CRISPR+editing.">Rapid interrogation of cancer cell of origin through CRISPR editing.</a> Proceedings of the National Academy of Sciences of the United States of America. 118 (32), 2110344118 (2021).</li></ol>

GEMM一直是从正常细胞开始的癌症建模的黄金标准，允许对潜在的致癌扰动（癌基因激活和/或肿瘤抑制因子的丢失）的后果进行严格测试。在这里，提供了一种快速有效的方案，通过对正常小鼠尿路上皮细胞进行 离体 基因编辑来生成膀胱癌类器官，这些细胞携带感兴趣的基因中的浮出等位基因。H&E和IHC染色表明，TKO类器官表现出与高级尿路上皮尿路上皮一致的组织学，具有鳞状分化和基底...

膀胱癌是男性中第四大常见癌症，每年在美国影响超过80，000人1。三十多年来，基于铂金的化疗一直是晚期膀胱癌患者的标准护理。最近美国食品和药物管理局（FDA）批准了免疫疗法（抗PD-1和抗PD-L1免疫检查点抑制剂），厄达非替尼（一种成纤维细胞生长因子受体抑制剂）和enfortumab vedotin（一种抗体 - 药物偶联物）2，3，4，2，4，从而彻底改变了膀胱癌治疗的前景。然而，没有临床批准的生物标志物可用于预测对化疗或免疫治疗的反应。迫切需要生成信息丰富的临床前模型，以提高对驱动膀胱癌进展的机制的理解，并为不同的治疗方式开发预测性生物标志物。
膀胱转化研究的一个主要障碍是缺乏临床前模型来概括人类膀胱癌的发病机制和治疗反应5，6。已经开发了多种临床前模型，包括 体外 2D模型（细胞系或有条件重编程细胞）， 体外 3D模型（类器官，3D打印）和 体内 模型（异种移植物，致癌物诱导，基因工程模型和患者来源的异种移植物）2，6。基因工程小鼠模型（GEMMs）可用于膀胱癌生物学中的许多应用，包括肿瘤表型分析，候选基因和/或信号通路的机制研究以及治疗反应的临床前评估6，7。GEMM可以利用位点特异性重组酶（Cre-loxP）来控制一个或多个肿瘤抑制基因中的遗传缺失。生成具有多个基因缺失的所需GEMM的过程非常耗时，费力且昂贵5.该方法的总体目标是开发一种快速有效的 离体 Cre递送方法，用于从携带三重 floxed 等位基因（Trp53，Pten 和 Rb1）的正常小鼠尿路上皮细胞建立膀胱三重敲除（TKO）模型8。 离体 方法的主要优点是快速的工作流程（1-2周而不是多年的小鼠育种）。本文描述了在免疫功能正常的C57 BL / 6J小鼠中收获具有浮选等位基因的正常尿路上皮细胞， 离体 腺病毒转导，类器官培养以及 体外 和 体内 表征的方案。该方法可进一步用于在含有任何絮状等位基因组合的免疫功能小鼠中产生临床相关的膀胱癌类器官。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>100 &#956;m sterile cell strainer</td>
			<td>Corning</td>
			<td>431752</td>
			<td></td>
		</tr>
		<tr>
			<td>1 mL syringe</td>
			<td>BD</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr>
			<td>25G 1.5 inches needle</td>
			<td>EXELINT International</td>
			<td>26406</td>
			<td></td>
		</tr>
		<tr>
			<td>Adenovirus (Ad5CMVCre High Titer, 1E11 pfu/ml)</td>
			<td>UI Viral Vector Core</td>
			<td>VVC-U of Iowa-5-HT</td>
			<td></td>
		</tr>
		<tr>
			<td>C57 BL/6J</td>
			<td>Jackson Lab</td>
			<td>000664</td>
			<td></td>
		</tr>
		<tr>
			<td>Charcoal-stripped FBS</td>
			<td>Gibco</td>
			<td>A3382101</td>
			<td></td>
		</tr>
		<tr>
			<td>Collagenase/hyaluronidase</td>
			<td>Stemcell Technologies</td>
			<td>07912</td>
			<td></td>
		</tr>
		<tr>
			<td>Dispase</td>
			<td>Stemcell Technologies</td>
			<td>07913</td>
			<td></td>
		</tr>
		<tr>
			<td>DPBS, 1x</td>
			<td>Corning</td>
			<td>21-031-CV</td>
			<td></td>
		</tr>
		<tr>
			<td>L-glutamine substitute (GlutaMAX)</td>
			<td>Gibco</td>
			<td>35-050-061</td>
			<td></td>
		</tr>
		<tr>
			<td>Mammary Epithelial Cell Growth medium</td>
			<td>Lonza</td>
			<td>CC-3150</td>
			<td></td>
		</tr>
		<tr>
			<td>Matrix extracts from Engelbreth&#8211;Holm&#8211;Swarm mouse sarcomas (Matrigel)</td>
			<td>Corning</td>
			<td>CB-40234</td>
			<td></td>
		</tr>
		<tr>
			<td>Monoclonal mouse anti-CK20</td>
			<td>DAKO</td>
			<td>M7019</td>
			<td>IF 1:100</td>
		</tr>
		<tr>
			<td>Monoclonal mouse anti-CK7</td>
			<td>Santa Cruz Biotechnology</td>
			<td>SC-23876</td>
			<td>IHC 1:50</td>
		</tr>
		<tr>
			<td>Monoclonal mouse anti-p63</td>
			<td>Abcam</td>
			<td>ab735</td>
			<td>IHC 1:100, IF 1:50</td>
		</tr>
		<tr>
			<td>Monoclonal mouse anti-Upk3</td>
			<td>Fitzgerald</td>
			<td>10R-U103A</td>
			<td>IHC 1:50</td>
		</tr>
		<tr>
			<td>Monoclonal mouse anti-Vimentin</td>
			<td>Santa Cruz Biotechnology</td>
			<td>SC-6260</td>
			<td>IF 1:100</td>
		</tr>
		<tr>
			<td>Monoclonal rat anti-CK8</td>
			<td>Developmental Studies Hybridoma Bank</td>
			<td>TROMA-I-s</td>
			<td>IF 1:100</td>
		</tr>
		<tr>
			<td>HERAcell vios 160i CO2 incubator</td>
			<td>Thermo Fisher</td>
			<td>51033557</td>
			<td></td>
		</tr>
		<tr>
			<td>Polyclonal chicken anti-CK5</td>
			<td>Biolegend</td>
			<td>905901</td>
			<td>IF 1:500</td>
		</tr>
		<tr>
			<td>Primocin</td>
			<td>InvivoGen</td>
			<td>ant-pm-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant enzyme of trypsin substitute (TrypLE Express Enzyme)</td>
			<td>Thermo Fisher</td>
			<td>12605036</td>
			<td></td>
		</tr>
		<tr>
			<td>Signature benchtop shaking incubator Model 1575</td>
			<td>VWR</td>
			<td>35962-091</td>
			<td></td>
		</tr>
		<tr>
			<td>Specimen Processing Gel (HistoGel)</td>
			<td>Thermo Fisher</td>
			<td>HG-4000-012</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical blade size 10</td>
			<td>Integra Miltex</td>
			<td>4-110</td>
			<td></td>
		</tr>
		<tr>
			<td>Sorvall T1 centrifuge</td>
			<td>Thermo Fisher</td>
			<td>75002383</td>
			<td></td>
		</tr>
		<tr>
			<td>Y-27632</td>
			<td>Selleckchem</td>
			<td>S1049</td>
			<td></td>
		</tr>
	</tbody>
</table>

ex vivo organoid model of adenovirus-cre mediated gene deletions in mouse urothelial cells

膀胱癌是一个研究不足的领域，特别是在基因工程小鼠模型（GEMMs）中。具有组织特异性Cre和loxP位点的自交GEMM一直是条件或诱导基因靶向的黄金标准。为了提供更快，更有效的实验模型，使用腺病毒Cre和携带肿瘤抑制因子Trp53，Pten和Rb1的多个loxP等位基因的正常尿路上皮细胞开发了离体类器官培养系统。正常的尿路上皮细胞与三重 floxed 小鼠的四个膀胱酶解脱（Trp53f / f：Ptenf / f：Rb1f / f）。尿路上皮细胞通过CMV启动子（Ad5CMVCre）驱动的腺病毒-Cre进行体外转导。将转导的膀胱类器官在体外和体内进行培养、繁殖和表征。PCR 用于确认 Trp53、Pten 和 Rb1 中的基因缺失。类器官的免疫荧光 （IF） 染色显示尿路上皮谱系标志物（CK5 和 p63）的阳性表达。将类器官皮下注射到宿主小鼠中以进行肿瘤扩张和连续传代。异种移植物的免疫组化（IHC）表现出CK7，CK5和p63的阳性表达以及CK8和Uroplakin 3的阴性表达。总之，腺病毒介导的基因从具有loxP位点工程化的小鼠尿路上皮细胞中缺失是快速测试已定义遗传改变的致瘤潜力的有效方法。

小鼠尿路上皮细胞中腺病毒-Cre介导的基因缺失的工作流程如图 1A所示。随附的视频演示了尿路上皮细胞 如何与膀胱 底分离，以及如何用Ad5CMVCre转导三重浮冰细胞。 图1A 显示，酶消化后，最小的粘膜下层和肌肉细胞被解离。为了确认腺病毒 - Cre递送的效率，用腺病毒Cre转导来自三重浮冰小鼠的解离的尿路上皮细胞与膜靶向tdTomato/膜定位增强的绿...

Watch this Scientific Journal Video about Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells at JoVE.com

Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells

该协议描述了小鼠尿路上皮类器官的产生和表征过程，这些类器官在感兴趣的基因中具有缺失。这些方法包括收获小鼠尿路上皮细胞，用腺病毒用CMV启动子驱动Cre表达的 离体 转导，以及 体外 和 体内 表征。

离体 小鼠尿路上皮细胞腺病毒-Cre介导的基因缺失的类器官模型

Bladder cancer is an understudied area, particularly in genetically engineered mouse models (GEMMs). Inbred GEMMs with tissue-specific Cre and loxP sites ...

我们开发了一种快速有效的方案，通过对正常小鼠尿路上皮细胞进行离体基因编辑来生成膀胱癌类器官，这些细胞携带感兴趣的基因中的浮出等位基因。这种离体方法具有一到两周的工作流程，而不是多年的小鼠育种，并且在Cre介导的基因缺失上的腺病毒转导中非常有效。首先，准备所有无菌仪器和试剂，包括剪刀，镊子，35毫升培养皿中的DPBS，70%乙醇，纱布和干净的纸巾。接下来，清洁解剖区域的所有表面，并用干净的纸巾覆盖解剖区域。将安乐死小鼠放在解剖区的纱布上，在小鼠腹部喷洒70%乙醇，然后使用无菌解剖剪刀，做一个下中线腹部切口，露出膀胱。现在，使用镊子抓住并轻轻拉起膀胱以识别双侧输尿管卵巢连接处。使用剪刀，沿着两个输尿管开口之间的边界线取出膀胱底，并将膀胱转移到含有2毫升DPBS的无菌培养皿中，然后去除脂肪和结缔组织，并将膀胱放入含有2毫升DPBS的新培养皿中。还要从零下80摄氏度的储存中取出腺病毒，并将其保存在冰上。对于细胞解离，按照手稿中所述准备所有无菌仪器和试剂，然后在生物安全柜中，将膀胱转移到35毫米培养皿中，并用2毫升无菌DPBS洗涤。将四只小鼠的膀胱组织收集到具有一毫升完整培养基的培养皿中，没有木炭剥离的FBS。接下来，使用手术刀片，将每个膀胱切成四个相等的碎片，然后使用镊子展开膀胱，将尿路上皮表面暴露在介质中。将气囊碎片和培养基转移到15毫升离心管中。用2毫升完整的培养基洗涤培养皿两次，没有木炭剥离的FBS，并将洗涤物加入离心管中，使总体积达到5毫升。接下来，加入胶原酶和透明质酸酶混合物，并将管水平置于37摄氏度的轨道振荡器培养箱中，以200 RPM的速度放置30分钟。组织消化后，将等体积的10%木炭剥离的FBS和DPBS加入管中，并以200倍g离心5分钟。弃去上清液后，向细胞沉淀中加入2毫升胰蛋白酶替代品并充分混合，然后将管在37摄氏度和5%二氧化碳下孵育4分钟。孵育后，用标准P1000吸头上下移液细胞10次，并加入5毫升10%木炭剥离的FBS和DPBS。通过100微米无菌细胞过滤器过滤解离的细胞，并将细胞悬浮液收集在50毫升管中。将细胞悬浮液转移到15毫升管中，并以200倍g离心5分钟。除去上清液后，将沉淀重悬于1毫升完整的培养基中。使用血细胞计数器计数细胞数，并将0.5次板10至第6个细胞放入24孔板的每个孔中，并加入0.5毫升新鲜完整的培养基。接下来，从零下20摄氏度的储存中取出Engelbreth-Holm-Swarm小鼠肉瘤细胞的基质提取物，并在冰上解冻。此外，在37摄氏度的细胞培养箱中预热6孔板。对于腺病毒转导，将2微升腺病毒加入24孔板中，并与解离的尿路上皮细胞充分混合。为了提高转导功效，通过将板以300倍g离心30分钟进行旋切，然后将板在37摄氏度和5%二氧化碳下孵育1小时。将细胞和培养基从孔中转移到15毫升管中，并以200倍g离心5分钟。弃去上清液后，加入50微升完全培养基，并与底部的细胞充分混合。接下来，加入140微升基质提取物并轻轻混合以避免气泡，然后通过将每个圆顶的50微升溶液快速加入预热的6孔板中来创建类器官圆顶。孵育板，让圆顶凝固5分钟。5分钟后，将6孔板倒置，继续凝固25分钟。孵育后，向每个孔中加入2.5毫升完整的培养基，并将板置于培养箱中进行类器官培养。在近100%的细胞中检测到绿色荧光蛋白，表明腺病毒转导效率高。苏木精和曙红染色及三重敲除类器官肿瘤的免疫组化反应显示，高级别尿路上皮癌形态，基底亚型标志物CK5和p63蛋白表达阳性。在未用腺病毒处理的尿路上皮细胞中检测到未重组的絮状等位基因，而在三重敲除类器官中观察到重组的等位基因。一般来说，需要2至3周才能形成2厘米的皮下肿瘤。体内异种移植物的三重敲除类器官的免疫组化结果显示CK7、CK5和p63蛋白的阳性表达。CK8和尿素蛋白的表达结果为阴性。间充质标志物vimentin仅在肿瘤囊或基质中检测到。小鼠膀胱解离不得超过30分钟。较长的解离时间可能会增加基质细胞的百分比，例如内皮细胞和成纤维细胞这种方法可以与细胞分选或细胞型特异性腺病毒相结合，以产生细胞类型的特定类器官，例如管腔与基底类器官的比较。

Research

JoVE Journal

Cancer Research

Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells

The Drosophila Imaginal Disc Tumor Model: Visualization and Quantification of Gene Expression and Tumor Invasiveness Using Genetic Mosaics

The Drosophila Imaginal Disc Tumor Model: Visualization and Quantification of Gene Expression and Tumor Invasiveness Using Genetic Mosaics

该<em&gt;果蝇</em&gt;成虫盘瘤模型：基因的表达与肿瘤侵袭性的可视化和定量遗传马赛克

Drosophila melanogaster has emerged as a powerful experimental system for functional and mechanistic studies of tumor development and progression in the ...

科研

癌症研究

Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer

在直肠癌原位小鼠模型中循环肿瘤细胞的分离

Despite the advantages of easy applicability and cost-effectiveness, subcutaneous mouse models have severe limitations and do not accurately simulate ...

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

体内在肿瘤微环境中 pH 值、 pO2、氧化还原状态、磷酸盐和谷胱甘肽浓度的 EPR 评价

This protocol demonstrates the capability of low-field electron paramagnetic resonance (EPR)-based techniques in combination with functional paramagnetic ...

A Preclinical Mouse Model of Osteosarcoma to Define the Extracellular Vesicle-mediated Communication Between Tumor and Mesenchymal Stem Cells

骨肉瘤前小鼠模型对肿瘤与间充质干细胞细胞外囊泡介导通信的定义

Within the tumor microenvironment, resident or recruited mesenchymal stem cells (MSCs) contribute to malignant progression in multiple cancer types. Under ...

Paramyxoviruses for Tumor-targeted Immunomodulation: Design and Evaluation Ex Vivo

肿瘤靶向免疫调节副病毒的设计与评价

Successful cancer immunotherapy has the potential to achieve long-term tumor control. Despite recent clinical successes, there remains an urgent need for ...

Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma

骨肉瘤的三维骨细胞外基质模型

Osteosarcoma (OS) is the most common and a highly aggressive primary bone tumor. It is characterized with anatomic and histologic variations along with ...

Evaluating the Differentiation Capacity of Mouse Prostate Epithelial Cells Using Organoid Culture

利用有机体培养评价小鼠前列腺皮细胞的分化能力

The prostate epithelium is comprised predominantly of basal and luminal cells. In vivo lineage tracing has been utilized to define the differentiation ...

An Ex Vivo Brain Slice Model to Study and Target Breast Cancer Brain Metastatic Tumor Growth

An Ex Vivo Brain Slice Model to Study and Target Breast Cancer Brain Metastatic Tumor Growth

一 种研究 并靶向乳腺癌脑转移性肿瘤生长的离体脑切片模型

Brain metastasis is a serious consequence of breast cancer for women as these tumors are difficult to treat and are associated with poor clinical ...

Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells

Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells

通过恶性周围神经鞘肿瘤细胞中絮状等位基因的 离体 消融来定义肿瘤发生的基因功能

The development of new drugs that precisely target key proteins in human cancers is fundamentally altering cancer therapeutics. However, before these ...

Maintaining Human Glioblastoma Cellular Diversity Ex vivo using Three-Dimensional Organoid Culture

Maintaining Human Glioblastoma Cellular Diversity Ex vivo using Three-Dimensional Organoid Culture

使用三维类器官培养物离 体 维持人胶质母细胞瘤细胞多样性

Glioblastoma (GBM) is the most commonly occurring primary malignant brain cancer with an extremely poor prognosis. Intra-tumoral cellular and molecular ...

离体 小鼠尿路上皮细胞腺病毒-Cre介导的基因缺失的类器官模型

离体小鼠尿路上皮细胞腺病毒-Cre介导的基因缺失的类器官模型