Name: studying the effects of tumor-secreted paracrine ligands on macrophage activation using co-culture with permeable membrane supports
Uploaded: 2019-11-28T13:30:00
Description: Watch this Scientific Journal Video about Studying the Effects of Tumor-Secreted Paracrine Ligands on Macrophage Activation using Co-Culture with Permeable Membrane Supports at JoVE.com

埃里克·乌比尔部分由美国癌症协会博士后奖学金（128770-PF-15-216-01-LIB）资助。这项工作得到了NIH（R01-CA205398）的资助和乳腺癌研究基金会（BCRF-18-041）对HSE的资助。

与采集和使用鼠腹巨噬细胞有关的所有程序在北卡罗来纳大学教堂山分校（UNC）进行，并经联合国哥伦比亚联合国委员会机构动物护理和使用委员会（IACUC）批准。
1. 巨噬菌体文化
注：本程序可以利用原发性围周巨噬细胞（如下所述）、骨髓衍生巨噬细胞或巨噬细胞系，如J774（ATCC）或RAW264（ATCC）。
<ol><li>收获围膜巨噬细胞如前所述16，17<...

<ol>
	<li>Freeman, G. 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=Engagement+of+the+PD-1+immunoinhibitory+receptor+by+a+novel+B7+family+member+leads+to+negative+regulation+of+lymphocyte+activation.">Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation.</a> Journal of Experimental Medicine. 192 (7), 1027-1034 (2000).</li><li>Agata, 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=Expression+of+the+PD-1+antigen+on+the+surface+of+stimulated+mouse+T+and+B+lymphocytes.">Expression of the PD-1 antigen on the surface of stimulated mouse T and B lymphocytes.</a> International Immunology. 8 (5), 765-772 (1996).</li><li>Dong, 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-associated+B7-H1+promotes+T-cell+apoptosis:+a+potential+mechanism+of+immune+evasion.">Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion.</a> Nature Medicine. 8 (8), 793-800 (2002).</li><li>Sznol, M., Chen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antagonist+antibodies+to+PD-1+and+B7-H1+(PD-L1)+in+the+treatment+of+advanced+human+cancer--response.">Antagonist antibodies to PD-1 and B7-H1 (PD-L1) in the treatment of advanced human cancer--response.</a> Clinical Cancer Research. 19 (19), 5542 (2013).</li><li>Kortylewski, 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=Regulation+of+the+IL-23+and+IL-12+balance+by+Stat3+signaling+in+the+tumor+microenvironment.">Regulation of the IL-23 and IL-12 balance by Stat3 signaling in the tumor microenvironment.</a> Cancer Cell. 15 (2), 114-123 (2009).</li><li>Halak, B. K., Maguire, H. C., Lattime, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor-induced+interleukin-10+inhibits+type+1+immune+responses+directed+at+a+tumor+antigen+as+well+as+a+non-tumor+antigen+present+at+the+tumor+site.">Tumor-induced interleukin-10 inhibits type 1 immune responses directed at a tumor antigen as well as a non-tumor antigen present at the tumor site.</a> Cancer Research. 59 (4), 911-917 (1999).</li><li>Wang, 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=Regulation+of+the+innate+and+adaptive+immune+responses+by+Stat-3+signaling+in+tumor+cells.">Regulation of the innate and adaptive immune responses by Stat-3 signaling in tumor cells.</a> Nature Medicine. 10 (1), 48-54 (2004).</li><li>Mazzoni, 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=Myeloid+suppressor+lines+inhibit+T+cell+responses+by+an+NO-dependent+mechanism.">Myeloid suppressor lines inhibit T cell responses by an NO-dependent mechanism.</a> The Journal of Immunology. 168 (2), 689-695 (2002).</li><li>Zea, A. 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=Arginase-producing+myeloid+suppressor+cells+in+renal+cell+carcinoma+patients:+a+mechanism+of+tumor+evasion.">Arginase-producing myeloid suppressor cells in renal cell carcinoma patients: a mechanism of tumor evasion.</a> Cancer Research. 65 (8), 3044-3048 (2005).</li><li>Steele, R. J., Eremin, O., Brown, M., Hawkins, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+high+macrophage+content+in+human+breast+cancer+is+not+associated+with+favourable+prognostic+factors.">A high macrophage content in human breast cancer is not associated with favourable prognostic factors.</a> British Journal of Surgery. 71 (6), 456-458 (1984).</li><li>Evans, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+T-+and+B+lymphocyte+responses+to+mitogens+by+tumor-associated+macrophages:+the+dependency+on+the+stage+of+tumor+growth.">Regulation of T- and B lymphocyte responses to mitogens by tumor-associated macrophages: the dependency on the stage of tumor growth.</a> Journal of Leukocyte Biology. 35 (6), 549-559 (1984).</li><li>Noy, R., Pollard, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor-associated+macrophages:+from+mechanisms+to+therapy.">Tumor-associated macrophages: from mechanisms to therapy.</a> Immunity. 41 (1), 49-61 (2014).</li><li>Pollard, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumour-educated+macrophages+promote+tumour+progression+and+metastasis.">Tumour-educated macrophages promote tumour progression and metastasis.</a> Nature Reviews Cancer. 4 (1), 71-78 (2004).</li><li>Renaud, J., Martinoli, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+Insert+Co-culture+System+of+Two+Cellular+Types+in+the+Absence+of+Cell-Cell+Contact.">Development of an Insert Co-culture System of Two Cellular Types in the Absence of Cell-Cell Contact.</a> Journal of Visualized Experiments. (113), e54356 (2016).</li><li>Dasari, S., Pandhiri, T., Haley, J., Lenz, D., Mitra, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Proximal+Culture+Method+to+Study+Paracrine+Signaling+Between+Cells.">A Proximal Culture Method to Study Paracrine Signaling Between Cells.</a> Journal of Visualized Experiments. (138), e58144 (2018).</li><li>Ubil, 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=Tumor-secreted+Pros1+inhibits+macrophage+M1+polarization+to+reduce+antitumor+immune+response.">Tumor-secreted Pros1 inhibits macrophage M1 polarization to reduce antitumor immune response.</a> Journal of Clinical Investigation. 128 (6), 2356-2369 (2018).</li><li>Ray, A., Dittel, B. N. <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+mouse+peritoneal+cavity+cells.">Isolation of mouse peritoneal cavity cells.</a> Journal of Visualized Experiments. (35), e1488 (2010).</li><li>Zaks-Zilberman, M., Zaks, T. Z., Vogel, S. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+proinflammatory+and+chemokine+genes+by+lipopolysaccharide+and+paclitaxel+(Taxol)+in+murine+and+human+breast+cancer+cell+lines.">Induction of proinflammatory and chemokine genes by lipopolysaccharide and paclitaxel (Taxol) in murine and human breast cancer cell lines.</a> Cytokine. 15 (3), 156-165 (2001).</li></ol>

这里介绍的共同培养测定是先前建立的测定的一种修改，允许研究免疫细胞激活的肿瘤分泌因子。虽然细胞-细胞接触已知能诱导免疫活动的变化，但肿瘤分泌配体调节免疫活化的能力却不太了解。我们描述了一种方法，与直接共培养不同，它可以用来询问肿瘤衍生的分泌因子如何影响免疫细胞激活，而不会混淆接触介导信号的性质。鉴于肿瘤分泌配体在免疫抑制中的潜在临床重要性，这种方法提...

最近的研究集中在癌细胞避免被免疫细胞检测、抑制局部免疫活化或在肿瘤微环境中产生耐受性肿瘤适应环境的能力。两大类肿瘤和免疫细胞相互作用已被描述，促进这些影响：接触介导相互作用或肿瘤分泌配体。肿瘤利用的接触介导免疫抑制的一个研究最充分和临床可治愈的机制是PD-L1的表达，它与T细胞上的PD-1相互作用，以抑制其激活和功能1，2。对由多种活化免疫细胞表达的干扰素伽马（IFN®）作出反应，肿瘤细胞可以增加PD-L1的表达，导致PD-1表达活性T细胞的耗尽，从而阻止它们有效消灭肿瘤细胞3。使用抗体来阻止PD-L1和PD-1之间的相互作用，目前用于治疗人类多种癌症类型4。鉴于这一临床成功和其他，肿瘤衍生免疫抑制机制的鉴定和靶向越来越受到重视。
除了抑制适应性免疫外，肿瘤还已知会分泌抑制先天免疫细胞的亲炎反应的因素。肿瘤衍生或肿瘤诱导的分泌物，包括IL-6、IL-10、VEGF、IL-23和菌群刺激因子（CSF-1），已被证明能抑制肿瘤微环境中自然杀伤细胞（NK）、粒细胞和树突状细胞的抗肿瘤反应5、6、7。肿瘤细胞也可以分泌因子，扭曲肿瘤微环境中骨髓衍生细胞的招募和分化，促进抑制T细胞活化8，9。
对肿瘤进展有深远影响的一种先天免疫细胞是巨噬细胞。多年来，肿瘤相关巨噬细胞（TAM）的存在一直被用作患者生存的负预后10。免疫抑制性TAM抑制肿瘤免疫细胞介导清除的概念是在40多年前提出的。最近，已经表明，巨噬菌体促进炎症反应可以降低调节，而亲肿瘤表型可以在肿瘤微环境中诱导。这些免疫抑制巨噬细胞有助于耐原反应，推动肿瘤进展和耐化学和免疫治疗12。鉴于巨噬细胞往往是肿瘤最丰富的白细胞之一，恢复其肿瘤特异性免疫活性是抗癌治疗物13的潜在靶点。
虽然肿瘤细胞和巨噬细胞之间的接触介导相互作用可以通过直接共培养进行建模，但使用渗透膜支持可以阐明哪些肿瘤分泌因子是免疫调节，而不会产生肿瘤-免疫细胞-细胞接触的潜在混淆影响。使用某种类似的方法，其他人已经证明了识别微胶质/神经元相互作用14以及肿瘤细胞与中皮细胞15分泌因子的潜力。我们还成功地利用这种共培养技术来描述肿瘤分泌蛋白Pros1的作用，这种蛋白质在用LPS和干扰素伽马16刺激围层巨噬细胞后，作为促进炎症基因表达的抑制者的作用。在这里，我们描述了一个简单明了的方法，可以用来询问肿瘤分泌因子如何影响巨噬菌体活化。

<table border="0" cellpadding="0" cellspacing="0" style="width:1419px;" width="1418">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:444px;">B16-F10</td>
			<td style="width:244px;">ATCC</td>
			<td style="width:344px;">ATCC CRL-6475</td>
			<td style="width:387px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">cDNA synthesis kit</td>
			<td>Promega</td>
			<td>A3500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DMEM/F12 media</td>
			<td>ThermoFisher Scientific- Gibco</td>
			<td>11320033</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fetal Bovine Serum</td>
			<td>Millipore</td>
			<td>TMS-013-B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">J774A.1</td>
			<td>ATCC</td>
			<td>ATCC TIB-67</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lipopolysaccharides from Escherichia coli O111:B4</td>
			<td>Sigma-Aldrich</td>
			<td>L5293-2ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Murine M-CSF</td>
			<td>Prospec</td>
			<td>CYT-439</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Penicillin-Streptomycin (10,000 U/mL)</td>
			<td>ThermoFisher Scientific- Gibco</td>
			<td>15140122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pros1 ELISA</td>
			<td>MyBioSource</td>
			<td>MBS2886720</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">RAW264.7</td>
			<td>ATCC</td>
			<td>ATCC TIB-71</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Recombinant Mouse IFN&#947;</td>
			<td>BioLegend</td>
			<td>575302</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sensimix SYBR Low-ROX kit</td>
			<td>Bioline</td>
			<td>QT625-05</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Transwell permeable supports</td>
			<td>Fisher Scientific</td>
			<td>07-200-170</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Trypsin-EDTA</td>
			<td>ThermoFisher Scientific- Gibco</td>
			<td>25200056</td>
			<td></td>
		</tr>
	</tbody>
</table>

studying the effects of tumor-secreted paracrine ligands on macrophage activation using co-culture with permeable membrane supports

肿瘤衍生的副苯并发信号是局部免疫抑制的一个被忽视的成分，可以导致一个允许的环境，持续癌症生长和转移。Paracrine 信号可能涉及不同细胞类型之间的细胞-细胞接触，例如 PD-L1 在肿瘤表面与 T 细胞表面的 PD-1 直接相互作用，或肿瘤细胞分泌配体以影响免疫细胞。在这里，我们描述了一种共培养方法，用于询问肿瘤分泌配体对免疫细胞（巨噬细胞）激活的影响。这种简单的程序利用市售的0.4μm聚碳酸酯膜渗透支架和标准组织培养板。在所述过程中，巨噬细胞在上腔培养，肿瘤细胞在下腔培养。0.4 μm 屏障的存在允许研究细胞间信号，而不会造成物理接触的混淆变量，因为两种细胞类型共享相同的介质，并暴露于副体配体。这种方法可以与其他方法相结合，例如巨噬细胞的基因改变（例如，从基因敲除小鼠分离）或肿瘤（例如，CRISPR介导的改变），以研究特定分泌因子和受体的作用。该方法还适用于标准分子生物学分析，如定量逆转录聚合酶链反应（qRT-PCR）或西方斑点分析，无需进行流动分拣来分离两个细胞群。酶相关免疫吸附测定（ELISAs）同样可用于测量分泌配体，以更好地了解多细胞类型上下文中细胞信号的动态相互作用。共同文化的时间也可以因研究时间调节事件而不同。这种共培养方法是一个强大的工具，有助于研究免疫环境中的肿瘤分泌信号。

为了确定肿瘤分泌配体对巨噬菌体极化的影响，采用了所述程序。在没有肿瘤细胞的情况下培养的食管巨噬细胞被用作阴性（未经治疗=最左侧）和阳性（IFN+和LPS刺激=从左至左为2）对照（图2A）。或者，围肠巨噬细胞与B16F10黑色素瘤肿瘤细胞（ATCC）共同培养（图1A）。电镀后立即，细胞要么使用IFN®和LPS进行治疗，要么不进行治疗。在24小时培?...

Watch this Scientific Journal Video about Studying the Effects of Tumor-Secreted Paracrine Ligands on Macrophage Activation using Co-Culture with Permeable Membrane Supports at JoVE.com

Studying the Effects of Tumor-Secreted Paracrine Ligands on Macrophage Activation using Co-Culture with Permeable Membrane Supports

在这里，我们提出了一种使用渗透膜支架的方法，以促进肿瘤细胞用于抑制免疫反应的非接触性对羟基苯甲酰二甲苯信号的研究。该系统有助于研究肿瘤分泌因子在抑制巨噬菌体活化中的作用。

研究肿瘤分泌性副体结扎对宏噬细胞活化的影响，使用可渗透膜支架共培养

Tumor-derived paracrine signaling is an overlooked component of local immunosuppression and can lead to a permissive environment for continued cancer ...

这种Transwell共培养方法用于研究肿瘤细胞用于抑制免疫反应的寄生虫信号。这种方法可用于发现新的分泌配体，以及其免疫抑制作用的机械基础。我们以前曾使用这种方法来显示肿瘤分泌因子如何抑制巨噬细胞亲炎基因转录。我们相信，这个工具在肿瘤衍生免疫抑制的研究中具有广泛的意义。该技术的主要优点之一是，它可用于研究肿瘤细胞和免疫细胞之间的寄生虫信号，而不用细胞间接触的潜在混淆变量。该平台还适用于各种分子生物学技术，用于下游分析。收获后，将内酮巨噬细胞直接板入0.4微米聚酯膜的上腔内插入共培养六井板。然后添加补充的DMEM到井，使顶部室包含一毫升和底部室包含1.5毫升。在37摄氏度下孵育三天，二氧化碳含量为5%。首先，按照 ATCC 推荐的组织培养方法，在各自的培养中培养可商用的肿瘤细胞。接下来，用PBS清洗粘附肿瘤细胞一次。在EDTA中加入0.05%的三辛，在37摄氏度下孵育，直到细胞分离。然后重新挂起包含 FBS 的介质中的单元格。使用血细胞计或细胞计数器定量细胞总数和离心机在220倍G五分钟颗粒。在离心过程中，从含有渗透膜支撑板的巨噬细胞的上腔和下腔中吸出介质，代之以新鲜介质。对于将镀膜的下腔室，用一毫升的介质而不是1.5毫升填充，以留下足够的体积来补充细胞。离心完成后，从颗粒肿瘤细胞中吸出介质，以每毫升30万个细胞的浓度重新悬浮补充DMEM中的细胞。将此细胞悬浮液的0.5毫升添加到所需油井的下腔中。为了诱导巨噬细胞亲炎基因表达，通过添加干扰素伽马，浓度为每毫升100毫微克，以每毫升50毫微克的浓度单独治疗或共同培养的巨噬细胞。根据需要改变培养中的治疗时间。巨噬细胞活化发生在两个小时内，一些肿瘤中位抑制发生八小时。24 小时共培养产生稳健且一致的肿瘤源抑制。作为一种负对照，培养巨噬细胞单独离开未经处理。作为一种阳性对照，用干扰素伽马和LPS以与样品相同的浓度单独培养的巨噬细胞进行治疗。在所需的孵育时间过后，根据测试需要，根据需要分离细胞分离或条件培养培养。为了分离细胞酸盐进行定量聚合酶链反应分析，从井的两个腔室中吸出介质，用两毫升PBS洗涤一次。将RNA解液缓冲液涂抹在含有巨噬细胞的顶室。之后，轻轻刮擦膜以释放细胞酸盐，然后根据RNA隔离试剂盒制造商的协议将其转移到收集管中进行进一步处理。此处描述的共培养方法涉及巨噬细胞和肿瘤细胞的培养，没有物理接触。在没有肿瘤细胞的情况下培养的性腺巨噬细胞被用作阴性与阳性对等对等。在B16F10肿瘤细胞的存在下共培养的佩罗酮性巨噬细胞，但没有激活刺激，不会增加亲炎相关基因的表达。这意味着肿瘤分泌配体要么不足以诱导亲炎基因表达本身，或者如果有免疫激活的肿瘤分泌物，准理体配体足以抑制它到其原生水平。这种共培养方法表明，当由干扰素伽马和LPS两极分化的巨噬细胞在肿瘤细胞存在的情况下培养时，当巨噬细胞系J774被围死巨噬细胞取代时，炎症相关基因表达的抑制减少了60%A可比水平的巨噬细胞蛋白炎基因抑制。先前的研究已经确定蛋白质S是一种肿瘤分泌因子，可以抑制巨噬细胞的活化，因此渗透膜支持共培养模型与lysA结合使用，在24小时后测定条件介质中蛋白质S的浓度。在来自干扰素伽马和LPS处理B16F10黑色素瘤细胞的有条件介质中，蛋白质S以每毫升约475毫微克的浓度表达。在相同条件下治疗的佩里托内巨噬细胞表示蛋白质S，每毫升约61毫微克。有趣的是，当共同培养时，干扰素伽马和LPS处理良好的蛋白质S的浓度约为每毫升86毫微克。这表明，当存在巨噬细胞时，巨噬细胞摄入肿瘤分泌蛋白质S或B16F10细胞分泌的蛋白质S的含量会大大减少。这些结果突出了巨噬细胞与肿瘤细胞共同培养时巨噬细胞活化和抛物线信号的深刻变化。Transwell 共培养方法可以回答与参数信号相关的各种问题。可以进行西方的印迹分析，以确定肿瘤分泌的蛋白质如何改变巨噬细胞中的各种信号通路。

Research

JoVE Journal

Cancer Research

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

Practical Considerations in Studying Metastatic Lung Colonization in Osteosarcoma Using the Pulmonary Metastasis Assay

应用肺转移法研究骨肉瘤转移性肺定植的实用思考

The pulmonary metastasis assay (PuMA) is an ex vivo lung explant and closed cell culture system that permits researchers to study the biology of lung ...

Characterization of Cell Membrane Extensions and Studying Their Roles in Cancer Cell Adhesion Dynamics

细胞膜延伸特性及其在癌细胞黏附动力学中的作用研究

The cell membrane&#39;s extension repertoire modulates various malignant behaviors of cancer cells, including their adhesive and migratory potentials. The ...

Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors

用基因编码荧光传感器评估毒理学氧化应激的成像方法

While oxidative stress is a commonly cited toxicological mechanism, conventional methods to study it suffer from a number of shortcomings, including ...

Dual Effects of Melanoma Cell-derived Factors on Bone Marrow Adipocytes Differentiation

黑色素瘤细胞衍生因子对骨髓脂肪分化的双重影响

The crosstalk between bone marrow adipocytes and tumor cells may play a critical role in the process of bone metastasis. A variety of methods are ...

Mesenchymal Stem Cell Isolation from Pulp Tissue and Co-Culture with Cancer Cells to Study Their Interactions

骨髓间充质干细胞在浆组织中的分离及与癌细胞联合培养研究其相互作用

Cancer as a multistep process and complicated disease is not only regulated by individual cell proliferation and growth but also controlled by tumor ...

Measuring Bone Remodeling and Recreating the Tumor-Bone Microenvironment Using Calvaria Co-culture and Histomorphometry

利用卡尔瓦里亚共同培养和教条法测量骨重塑和再造肿瘤-骨微环境

Bone is a connective tissue constituted of osteoblasts, osteocytes, and osteoclasts and a mineralized extracellular matrix, which gives it its strength ...

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 ...

Characterization of the Effects of Migrastatic Inhibitors on 3D Tumor Spheroid Invasion by High-resolution Confocal Microscopy

通过高分辨率共聚焦显微镜对3D肿瘤球体入侵的影响表征

Drug discovery and development in cancer research is increasingly being based on drug screens in a 3D format. Novel inhibitors targeting the migratory and ...

Co-culture of Glioblastoma Stem-like Cells on Patterned Neurons to Study Migration and Cellular Interactions

胶质母细胞瘤干细胞在模式神经元上共同培养，以研究迁移和细胞相互作用

Glioblastomas (GBMs), grade IV malignant gliomas, are one of the deadliest types of human cancer because of their aggressive characteristics. Despite ...