Name: isolation and expansion of cytotoxic cytokine-induced killer t cells for cancer treatment
Uploaded: 2020-01-24T14:00:00
Description: Watch this Scientific Journal Video about Isolation and Expansion of Cytotoxic Cytokine-induced Killer T Cells for Cancer Treatment at JoVE.com

这项研究得到了中国医科大学附属医院（DMR-Cell-1809）的支持。

临床规程按照中国医科大学机构审查委员会和医院研究伦理委员会的指导方针进行和批准。外周血标本是在健康志愿者知情同意后采集的。
1. 材料准备
<ol><li>储存试剂、抗体和化学品，如材料安全数据表 （MSDS） 所示。将药物或细胞因子溶解在溶剂中作为库存溶液，然后等分储存在-20°C或-80°C。 注：材料准备的详细信息见材料表。</li>
</ol>2. ...

<ol>
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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=Cytokine-induced+killer+(CIK)+cells+in+cancer+immunotherapy:+report+of+the+international+registry+on+CIK+cells+(IRCC).">Cytokine-induced killer (CIK) cells in cancer immunotherapy: report of the international registry on CIK cells (IRCC).</a> Journal of Cancer Research and Clinical Oncology. 141 (5), 839-849 (2015).</li><li>Rutella, 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=Adoptive+immunotherapy+with+cytokine-induced+killer+cells+generated+with+a+new+good+manufacturing+practice-grade+protocol.">Adoptive immunotherapy with cytokine-induced killer cells generated with a new good manufacturing practice-grade protocol.</a> Cytotherapy. 14 (7), 841-850 (2012).</li><li>Nausch, N., 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=NKG2D+ligands+in+tumor+immunity.">NKG2D ligands in tumor immunity.</a> Oncogene. 27 (45), 5944-5958 (2008).</li><li>Gammaitoni, 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=Effective+activity+of+cytokine-induced+killer+cells+against+autologous+metastatic+melanoma+including+cells+with+stemness+features.">Effective activity of cytokine-induced killer cells against autologous metastatic melanoma including cells with stemness features.</a> Clinical Cancer Research. 19 (16), 4347-4358 (2013).</li><li>Rettinger, 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=The+cytotoxic+potential+of+interleukin-15-stimulated+cytokine-induced+killer+cells+against+leukemia+cells.">The cytotoxic potential of interleukin-15-stimulated cytokine-induced killer cells against leukemia cells.</a> Cytotherapy. 14 (1), 91-103 (2012).</li><li>Narayan, 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=Donor-derived+cytokine-induced+killer+cell+infusion+as+consolidation+after+nonmyeloablative+allogeneic+transplantation+for+myeloid+neoplasms.">Donor-derived cytokine-induced killer cell infusion as consolidation after nonmyeloablative allogeneic transplantation for myeloid neoplasms.</a> Biology of Blood and Marrow Transplantation. 19, 1083 (2019).</li><li>Castiglia, 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=Cytokines+induced+killer+cells+produced+in+good+manufacturing+practices+conditions:+identification+of+the+most+advantageous+and+safest+expansion+method+in+terms+of+viability,+cellular+growth+and+identity.">Cytokines induced killer cells produced in good manufacturing practices conditions: identification of the most advantageous and safest expansion method in terms of viability, cellular growth and identity.</a> Journal of Translational Medicine. 16 (1), 237 (2018).</li><li>Bonanno, G., 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=Thymoglobulin,+interferon-&#947;+and+interleukin-2+efficiently+expand+cytokine-induced+killer+(CIK)+cells+in+clinical-grade+cultures.">Thymoglobulin, interferon-&#947; and interleukin-2 efficiently expand cytokine-induced killer (CIK) cells in clinical-grade cultures.</a> Journal of Translational Medicine. 8, 129 (2010).</li><li>Iudicone, 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=Interleukin-15+enhances+cytokine+induced+killer+(CIK)+cytotoxic+potential+against+epithelial+cancer+cell+lines+via+an+innate+pathway.">Interleukin-15 enhances cytokine induced killer (CIK) cytotoxic potential against epithelial cancer cell lines via an innate pathway.</a> Human Immunology. 77 (12), 1239-1247 (2016).</li><li>Liu, 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=Phenotypic+characterization+and+anticancer+capacity+of+CD8++cytokine-induced+killer+cells+after+antigen-induced+expansion.">Phenotypic characterization and anticancer capacity of CD8+ cytokine-induced killer cells after antigen-induced expansion.</a> PLoS One. 12 (4), 0175704 (2017).</li><li>Chen, 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=Cytokine-induced+killer+cells+as+a+feasible+adoptive+immunotherapy+for+the+treatment+of+lung+cancer.">Cytokine-induced killer cells as a feasible adoptive immunotherapy for the treatment of lung cancer.</a> Cell Death &amp; Disease. 9 (3), 366 (2018).</li><li>Tario, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+cell+proliferation+by+dye+dilution:+considerations+for+probe+selection.">Monitoring cell proliferation by dye dilution: considerations for probe selection.</a> Methods in Molecular Biology. 1678, 249-299 (2018).</li><li>Last'ovicka, 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=Assessment+of+lymphocyte+proliferation:+CFSE+kills+dividing+cells+and+modulates+expression+of+activation+markers.">Assessment of lymphocyte proliferation: CFSE kills dividing cells and modulates expression of activation markers.</a> Cellular Immunology. 256 (1-2), 79-85 (2009).</li><li>Yoshida, 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=Characterization+of+natural+killer+cells+in+tamarins:+a+technical+basis+for+studies+of+innate+immunity.">Characterization of natural killer cells in tamarins: a technical basis for studies of innate immunity.</a> Frontiers in Microbiology. 1, 1-9 (2010).</li></ol>

所述方法是一种快速、方便、可靠的方案，用于从健康献血者全血样本中分离和扩增细胞毒性细胞因子（CIK）T细胞。它还显示了CIK对白血病（K562）和卵巢癌细胞（OC-3）的细胞毒性作用，使用流式细胞仪设置和跟踪（CS & T）系统。CIK细胞可以通过GMP级细胞因子和无血清培养基在良好的生产实践（GMP）条件下诱导和扩大，以进一步临床输注11。然而，CIK诱导和扩张的功效表现出个?...

细胞毒性T淋巴细胞是一种特定的免疫效应细胞群，用于调节对癌症的免疫反应。包括淋巴素激活杀伤性（LAK）细胞、肿瘤渗透淋巴细胞（TLL）、自然杀伤细胞（NK）细胞、βT细胞和细胞因子诱导杀伤细胞（CIK）细胞在内的几种效应细胞群已开发用于采用T细胞治疗（ACT）目的1。人们对CIK细胞的兴趣日益浓厚，因为它们代表了细胞因子引起的细胞毒性细胞群的混合物，这些细胞群从自体外周血单核细胞（PMBC）2中扩展。
淋巴原细胞、骨髓细胞和淋巴细胞的无控制生长导致三种主要类型的血癌（即白血病、淋巴瘤和骨髓瘤）、实体肿瘤，包括癌瘤（如肺癌、胃癌、宫颈癌）和肉瘤等癌症3。CIK细胞是细胞群的混合物，表现出广泛的主要组织相容性复合物（MHC）-不受限制的抗肿瘤活性，从而有望治疗血液学和晚期肿瘤4，5，6，7。CIK细胞包括细胞的组合，包括T细胞（CD3+CD56+）、NK-T细胞（CD3+CD56+）和NK细胞（CD3+CD56+）。通过使用固定的时间表来添加IFN-α、抗CD3抗体和IL-2，优化CIK诱导协议，导致CIK细胞8的膨胀。CIK细胞对癌细胞的细胞毒性主要取决于NK组2成员D（C型叶酸样受体家族成员）对肿瘤细胞的NKG2D配体和穿孔素介导途径9的活性。临床前研究结果表明，IL-15刺激CIK细胞在体外诱导强效细胞毒性对原发性及急性骨髓性白血病细胞系，对正常PBMC和成纤维细胞9表现出较低的感化活性。最近，在II期临床研究中，一次性健康供体衍生CIK（1 x 108/kgCD3+细胞）输注作为骨髓异体肿瘤治疗非骨髓异体移植的合并结果发表于10。
在本研究中，我们开发了由IFN-α、IL-1+、抗CD3抗体和IL-2添加到造血细胞培养基中增加CIK产量的优化细胞培养公式，并研究了CIK细胞对人体慢性的细胞毒性作用骨髓性白血病（K562）细胞和卵巢癌（OC-3）细胞。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>7-Amino Actinomycin D</td>
			<td>BD</td>
			<td>559925</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>APC Mouse Anti-Human CD56 antibody</td>
			<td>BD</td>
			<td>555518</td>
			<td>B159</td>
			<td></td>
		</tr>
		<tr>
			<td>APC Mouse IgG1, &#954; Isotype Control</td>
			<td>BD</td>
			<td>555751</td>
			<td>MOPC-21</td>
			<td></td>
		</tr>
		<tr>
			<td>BD FACSCanto II Flow Cytometer</td>
			<td>BD</td>
			<td>338962</td>
			<td></td>
			<td>SN: R33896202856</td>
		</tr>
		<tr>
			<td>Carboxyfluorescein diacetate succinimidyl ester (CFSE)</td>
			<td>BD</td>
			<td>565082</td>
			<td></td>
			<td>Reconstritution of CFSE dye (500 mg) with 90 mL of DMSO</td>
		</tr>
		<tr>
			<td>D-(+)-Glucose solution</td>
			<td>SIGMA</td>
			<td>G8644</td>
			<td></td>
			<td>For K562 cell culture. Add 12.5 mL to 500 mL of complete medium</td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s Modified Eagle Medium/F12</td>
			<td>HyClone</td>
			<td>SH30023.02</td>
			<td></td>
			<td>Basal medium for OC-3 cell culture</td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>HyClone</td>
			<td>SH30084.03</td>
			<td></td>
			<td>For K562 and OC-3 cell culture. Complete medium contains 10% of FBS</td>
		</tr>
		<tr>
			<td>Ficoll-Paque Plus</td>
			<td>GE Healthcare Life Sciences</td>
			<td>71101700-EK</td>
			<td></td>
			<td>Density gradient solution</td>
		</tr>
		<tr>
			<td>FITC Mouse Anti-Human CD3 antibody</td>
			<td>BD</td>
			<td>555332</td>
			<td>UCHT1</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC Mouse IgG1, &#954; Isotype Control</td>
			<td>BD</td>
			<td>555748</td>
			<td>MOPC-21</td>
			<td></td>
		</tr>
		<tr>
			<td>Human anti-CD3 mAb</td>
			<td>TaKaRa</td>
			<td>T210</td>
			<td>OKT3</td>
			<td>Add 2.5 mL of stock (1 mg/1 mL) to 50 mL of Induction medium. Storage stock at -80 &#176;C</td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin</td>
			<td>Gibco</td>
			<td>15140122</td>
			<td></td>
			<td>Add 5 mL of stock (10,000 U/mL) to 500 mL of complete medium. Storage stock at 4 &#176;C.</td>
		</tr>
		<tr>
			<td>Proleukin</td>
			<td>NOVARTIS</td>
			<td></td>
			<td></td>
			<td>Reconstitution of Proleukin Powder (22x106 IU) with 1.2 mL of sterile water and add 2.7 mL to 50 mL of Induction medium. Storage stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>Recombinant Human Interferon-gamma</td>
			<td>CellGenix</td>
			<td>1425-050</td>
			<td></td>
			<td>Reconstitution of rh IFN-g (5x105 IU/50 &#181;g) with 200 &#181;L of sterile water and add 20 mL to 50 mL of Induction medium. Storage stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>Recombinant Human Interleukin-1 alpha</td>
			<td>PEPROTECH</td>
			<td>200-01A</td>
			<td></td>
			<td>Reconstitution rh IL-1&#945; (10 &#181;g) with 1 mL of sterile water and add 5 mL to 50 mL of Induction medium. Storage stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>RPMI1640 medium</td>
			<td>Gibco</td>
			<td>11875-085</td>
			<td></td>
			<td>Basal medium for K562 cell culture. Storage stock at 4 &#176;C</td>
		</tr>
		<tr>
			<td>Sigma 3-18K Centrifuge</td>
			<td>Sigma</td>
			<td>10295</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>TrypLE Express Enzyme</td>
			<td>Gibco</td>
			<td>12605028</td>
			<td></td>
			<td>Cell dissociation enzyme; For deattachment of adheren cells. Storage at room temperature</td>
		</tr>
		<tr>
			<td>X-VIVO 15 medium</td>
			<td>Lonza</td>
			<td>04-418Q</td>
			<td></td>
			<td>Basal medium for PBMC and CIK cells. Storage at 4 &#176;C</td>
		</tr>
	</tbody>
</table>

isolation and expansion of cytotoxic cytokine-induced killer t cells for cancer treatment

采用的细胞免疫疗法侧重于通过免疫系统恢复癌症识别，并改善有效的肿瘤细胞杀灭。据报道，细胞因子诱导杀手（CIK）T细胞疗法对癌细胞产生显著的细胞毒性作用，并减少手术、放疗和化疗在癌症治疗中的不利影响。CIK可以从外周血单核细胞（PBMC）、骨髓和脐带血中提取。CIK细胞是具有CD3+CD56+和天然杀伤性（NK）型型特征的T细胞的异构亚群，包括主要组织相容性复合物（MHC）-无限制抗肿瘤活性。本研究描述了一种合格的、临床上适用的流式细胞学方法，用于定量PBMC衍生CIK细胞的细胞解解能力，对抗血液学和固体癌细胞。在细胞解物测定中，CIK细胞与预染色靶肿瘤细胞以不同比例共同孵育。潜伏期后，目标细胞的数量由核酸结合染色确定，以检测死细胞。该方法适用于研究和诊断应用。CIK 细胞具有强效细胞毒性，通过基于 CS 和 T 的流式细胞测定系统进行临床前评估后，可探索为癌症治疗的替代策略。

本方案的目的是从外周血单核细胞中分离和扩大细胞因子诱导杀伤性（CIK）T细胞，并分别评估CIK对血液恶性肿瘤和固体癌细胞的细胞毒性作用。CD3/CD56 识别确定了 CIK 的归纳。图 1A显示了 CIK 感应和扩展的协议。图1B显示了分析来自健康捐赠者的CD3+CD56+T细胞亚群的门控策略的代表性结果。图1</str...

Watch this Scientific Journal Video about Isolation and Expansion of Cytotoxic Cytokine-induced Killer T Cells for Cancer Treatment at JoVE.com

Isolation and Expansion of Cytotoxic Cytokine-induced Killer T Cells for Cancer Treatment

在这里，我们提出一个协议，以执行外周血单核细胞衍生细胞诱导CD3 + CD56+杀伤细胞的分离和膨胀，并说明其细胞毒性对血液和固体癌细胞的影响，通过使用体外诊断流细胞测定系统。

细胞毒性细胞因子诱导的杀伤性T细胞的分离和扩张，用于癌症治疗

Adoptive cellular immunotherapy focuses on restoring cancer recognition via the immune system and improves effective tumor cell killing. Cytokine-induced ...

我们优化了一种高素质和临床适用的方法，用于扩大PBMC衍生细胞因子诱导的杀手T细胞，并评估其细胞毒性对癌症。该技术的优点是为技术人员和临床医生提供一个标准的操作程序，以量化细胞因子诱导的杀手细胞在科学研究和临床评估中的效力。从文本协议中描述的 CIK 细胞制备开始此过程，然后用 10 毫升无菌 PBS 清洗 CIK 细胞。在300倍G和18至20摄氏度下离心10分钟。吸升液，用五毫升PBS重新暂停细胞。使用 trypan 蓝色排除测定计算细胞数并测试细胞生存能力。将 CIK 细胞分成六个灭菌 1.5 毫升管，密度约为每毫升 PBS 5 到 100 万个细胞。标记和对待文本协议中详细显示的单元格。用一毫升消毒移液器轻轻上下移液三次，轻轻地将 CIK 细胞与抗体混合。在黑暗中的室温下孵育细胞15分钟。在300倍G和18至20摄氏度下离心管10分钟。离心后，吸出上经剂，然后用一毫升PBS重新暂停细胞颗粒，然后用一毫升无菌移液器轻轻移液细胞上下至少三次。将细胞悬浮液转移到无菌的五毫升聚苯乙烯圆形底部管中，用细胞过滤器盖轻轻移液通过盖，然后将管放在旋转木马上。打开流式细胞仪分析软件并创建一个实验文件夹，然后单击新的试样按钮，向实验中添加试样和管。按文本协议中列出的管道命名。要为 CIK 细胞群创建散射门控系统，请先选择管一，然后单击点图按钮以创建 FSCA SSCA 图。在 FSCA 阈值大于 5000 的整个细胞群体上绘制矩形门，以排除细胞碎片。选择新点图的 FSCA FSCH 参数，并在所有单个单元格周围绘制多边形门。选择计数与 FITC 结合 CD3 和计数与 APC 结合 CD56 参数的新直方图图。选择用于新点图的 FITC 结合 CD3 与 APC 结合 CD56 参数，并绘制四个象限门来定义四个子组。记录每个标本中2万个单细胞的数据。单击加载示例按钮，首先分析空白控件示例。使用 CD56 和 CD3 通道参数确定整个 CIK 信元组。打开包含单个样本统计值的文件，分析 CIK 细胞量，然后重新打印到分析文件中。为了进行细胞毒性测定，共培养CIK和人类慢性骨髓性白血病K562细胞，将一毫升K562细胞加入六井板中，每毫升密度为500万细胞，然后将一毫升具有或没有CIK细胞的基底培养基加入文本协议中列出的六井板。通过轻轻上下移液至少三次，混合细胞悬浮液。将盘子放在培养箱中 24 小时。现在，通过添加一毫升基础培养基基，有或没有CIK细胞到六井板，如文本协议中列出的联合培养CIK和卵巢OC-3细胞。通过轻轻上下移液至少三次，混合细胞悬浮液。将盘子放入孵化器 24 小时。孵育后，将 CIK K562 细胞悬浮液直接收获到 15 毫升无菌管中。要从 CIK OC-3 组收获悬浮细胞和粘附细胞，请先将细胞悬浮液转移到 15 毫升无菌管中。用一毫升的杀菌PBS洗井，收集PBS，并加入管中，然后加入5毫升的细胞分离酶溶液，在37摄氏度下孵育5分钟。将溶液的一毫升从同一管添加到相应的井中，然后用一毫升无菌移液器上下移液，轻轻混合细胞。收集同一管中的所有细胞。在300次G下离心10分钟。吸升液，将细胞重新在一毫升无菌PBS中。在300次G下将细胞培养10分钟，然后吸进上那常物，并在100微升无菌PBS中重新产生细胞。在细胞悬浮液中加入5微升的7-AAD染料。用一毫升无菌移液器上下移液至少三次，轻轻混合细胞。孵育10分钟，在分析前在黑暗中离开。要执行细胞解能力测定，混合细胞悬浮液，并重复流式细胞仪的准备。单击新的试样按钮，向实验中添加试样和管。按文本协议中列出的管道命名。要为细胞解分析创建散射门控系统，请先选择管一，然后单击点图按钮以创建 FSCA SSCA 图。在 FSCA 阈值大于 50，000 的所有事件上绘制一个矩形门，以排除细胞碎片。选择新点图的 SSCA CFSE 参数，然后为新点图选择 7-AAD CFSE 参数，并绘制四个象限门来定义四个子填充。单击加载示例按钮，首先分析空白控件示例。调整 SSCA 和 FSCA 的电压。使用 CFSE 和 7-AAD 通道参数识别死细胞组。记录每个样本中超过 20，000 个 CFSE 阳性细胞的数据。打开包含每个样本的统计值的文件，分析不可行的细胞群，并将数据导出到分析文件中。通过三个个体的CIK比例统计分析，显示了用于分析健康捐献者CD-3阳性CD56阳性T细胞亚细胞的浇注策略的代表性结果。CD-3阳性CD56阳性细胞比例在扩张14天后显著增加。这一点通过原始PBMC的65%明显，这增加到27.4%的CIK细胞收获的第14天。在这个培养系统中，与原始数量的PBMC相比，CIK细胞产生大约半百倍的变化。 K562细胞被染色为非荧光染料CFSE，这种染料由细胞内在可行细胞内切割，成为一种高度荧光染料。说明了 CIK 和 CFSE 阳性细胞的大小和粒度。CFSE染色K562细胞与CIK细胞以0比1、5比1和10比1的比例进行共治疗。这些细胞被染色7-AAD染料作为死细胞排除的生存能力探针。对CFSE阳性K562细胞的7-AAD阳性细胞进行了全部评估。CFSE染色的OC-3细胞与CIK细胞以10比1的比例进行共治疗。在24小时的孵育后，在这里证明了CIK对OC-3细胞的明显细胞毒性。据报道，CIK T细胞对癌细胞产生显著的细胞毒性作用，减少手术、放疗和化疗在癌症治疗中的不利影响。免疫疗法是许多癌症的一种有希望的治疗方法。CIK可以通过在GMP级条件下孵化患者衍生的PMBC，在体外有效扩展，以进一步进行自体输注。按照此协议，我们可以在GTP或GMP级设施中执行免疫细胞治疗，以进一步临床癌症治疗。

Research

JoVE Journal

Cancer Research

Generation and Isolation of Cell Cycle-arrested Cells with Complex Karyotypes

复杂核型细胞周期捕获细胞的生成与分离

Chromosome mis-segregation leads to aneuploidy, a condition in which cells harbor an imbalanced chromosome number. Several lines of evidence strongly ...

科研

癌症研究

A Syngeneic Mouse B-Cell Lymphoma Model for Pre-Clinical Evaluation of CD19 CAR T Cells

同源小鼠 B 细胞淋巴瘤模型对 CD19 轿车 T 细胞的临床前评价

The astonishing clinical success of CD19 chimeric antigen receptor (CAR) T-cell therapy has led to the approval of two second generation chimeric antigen ...

Preparation of Exosomes for siRNA Delivery to Cancer Cells

用于向癌细胞输送 sirna 的外质体的制备

Extracellular vesicles, in particular exosomes, have recently gained interest as novel drug delivery vectors due to their biological origin, abundance, ...

Manufacturing Chimeric Antigen Receptor (CAR) T Cells for Adoptive Immunotherapy

Manufacturing Chimeric Antigen Receptor CAR T Cells for Adoptive Immunotherapy

制造用于采用免疫疗法的奇美抗原受体（CAR）T细胞

Adoptive immunotherapy holds promise for the treatment of cancer and infectious disease. We describe a simple approach to transduce primary human T cells ...

Using Human Induced Pluripotent Stem Cells for the Generation of Tumor Antigen-specific T Cells

利用人类诱导多能干细胞生成肿瘤抗原特异性T细胞

The generation and expansion of functional T cells in vitro can lead to a broad range of clinical applications. One such use is for the treatment of ...

Generation of Orthotopic Pancreatic Tumors and Ex vivo Characterization of Tumor-Infiltrating T Cell Cytotoxicity

Generation of Orthotopic Pancreatic Tumors and Ex vivo Characterization of Tumor-Infiltrating T Cell Cytotoxicity

正交胰腺肿瘤的产生与肿瘤渗透T细胞细胞毒性的外体特征

In vivo models of pancreatic cancer provide invaluable tools for studying disease dynamics, immune infiltration and new therapeutic strategies. The ...

Isolating Human Peripheral Blood Mononuclear Cells and CD4+ T cells from S&#233;zary Syndrome Patients for Transcriptomic Profiling

Isolating Human Peripheral Blood Mononuclear Cells and CD4+ T cells from SÃƒÆ’Ã‚Â©zary Syndrome Patients for Transcriptomic Profiling

从Sézary综合征患者中分离人外周血单核细胞和CD4 + T细胞以进行转录组学分析

Cutaneous T-cell lymphomas (CTCL) are derived from the transformation and uncontrolled proliferation of mature skin-homing T cells, and mycosis fungoides ...

Natural Killer (NK) and CAR-NK Cell Expansion Method using Membrane Bound-IL-21-Modified B Cell Line

Natural Killer NK and CAR-NK Cell Expansion Method using Membrane Bound-IL-21-Modified B Cell Line

使用膜结合IL-21修饰B细胞系的自然杀伤（NK）和CAR-NK细胞扩增方法

Chimeric antigen receptor (CAR)-modified immune cell therapy has become an emerging treatment for cancers and infectious diseases. NK-based immunotherapy, ...

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 ...

Generation and Expansion of Primary, Malignant Pleural Mesothelioma Tumor Lines

Current methodologies for the expansion of primary tumor cell lines from rare tumor types are lacking. This protocol describes methods to expand primary ...