Methods in cell biology最新文献_第4页

Cell culture techniques for cancer research. 用于癌症研究的细胞培养技术。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2025-03-18 DOI: 10.1016/bs.mcb.2025.02.023

Preeti Jain, Nitika Joshi, Sadhna Aggarwal

引用次数: 0

Liquid biopsy and circulating tumor cell analysis. 液体活检和循环肿瘤细胞分析。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2025-03-11 DOI: 10.1016/bs.mcb.2025.02.019

Wasiur Rahman Choudhury, Rama Rao Damerla, K Devaraja

引用次数: 0

Ex vivo assessment of human neutrophil motility and migration. 人中性粒细胞运动和迁移的离体评估。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-19 DOI: 10.1016/bs.mcb.2024.10.008

Noor A M Bakker, Claudia Burrello, Karin E de Visser

引用次数: 0

In vitro ILC differentiation from human HSCs. 人造血干细胞的体外ILC分化。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-19 DOI: 10.1016/bs.mcb.2024.10.004

Silvia Santopolo, Cecilia Ciancaglini, Francesca Romana Mariotti, Lorenzo Moretta, Linda Quatrini

{"title":"In vitro ILC differentiation from human HSCs.","authors":"Silvia Santopolo, Cecilia Ciancaglini, Francesca Romana Mariotti, Lorenzo Moretta, Linda Quatrini","doi":"10.1016/bs.mcb.2024.10.004","DOIUrl":"10.1016/bs.mcb.2024.10.004","url":null,"abstract":"The Innate Lymphoid Cells (ILCs) are a family of innate immune cells composed by the Natural Killer (NK) cells and the helper ILCs (hILCs) (ILC1, ILC2, ILC3), both developing from a common ILC precursor (ILCP) derived from hematopoietic stem cells (HSCs). A correct ILC reconstitution is crucial, particularly in patients receiving HSC transplantation (HSCT), the only therapeutic option for many adult and pediatric high-risk hematological malignancies. Indeed, mainly thanks to their cytotoxic activity, NK cells have a strong Graft-versus-Leukemia (GvL) effect. On the other hand, hILCs, that are mainly tissue resident, are involved in tissue repair and homeostasis, Graft-versus-Host Disease (GvHD) prevention and immune response to infections. Unlike NK cell development, hILC-poiesis is still poorly characterized in humans. Here, we provide a protocol for the in vitro ILC differentiation from healthy donor peripheral blood-derived CD34+ HSCs. This could represent a useful model to dissect the molecular mechanisms by which the distinct ILC subsets are generated from ILCP leading to the development of novel strategies to improve the HSCT clinical outcome.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"191 ","pages":"41-57"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143008130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Retrovirus-based manufacturing of chimeric antigen receptor-modified T cells for cancer therapy research. 基于逆转录病毒制造嵌合抗原受体修饰的T细胞用于癌症治疗研究。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-19 DOI: 10.1016/bs.mcb.2024.10.017

Sophia Stock, Luisa Fertig, Vivien Doreen Menkhoff, Thaddäus Strzalkowski, Manuel Caruso, Sebastian Kobold

{"title":"Retrovirus-based manufacturing of chimeric antigen receptor-modified T cells for cancer therapy research.","authors":"Sophia Stock, Luisa Fertig, Vivien Doreen Menkhoff, Thaddäus Strzalkowski, Manuel Caruso, Sebastian Kobold","doi":"10.1016/bs.mcb.2024.10.017","DOIUrl":"10.1016/bs.mcb.2024.10.017","url":null,"abstract":"Treatment with autologous chimeric antigen receptor (CAR)-modified T cells can achieve outstanding clinical response rates in heavily pretreated patients with B and plasma cell malignancies. However, relapses occur, and they limit the efficacy of this promising treatment approach. The complex GMP-compliant production and high treatment costs cause that CAR T cells cannot yet be used in a broad population. Among others, CAR T cell therapy has evolved regarding vector design and manufacturing process. Optimal production of CAR T cells is not yet defined, far from being standardized. Quality, cellular composition and immunophenotype of the administered CAR T cells are influenced by the manufacturing protocol and therefore play a crucial role for therapeutic success. For the gene transfer, viral and non-viral strategies are available. Retrovirus-based protocols for CAR T cell production offer advantages in terms of stable gene integration, sufficient transduction efficiency, proven clinical success, and scalability. Here, we detail a retrovirus-based generation protocol of human CAR-modified T cells for experimental immunotherapeutic treatment of cancer cells. For the CAR generation, HEK-293-based packaging cell lines, CD3+ selection, CD3/CD28-coated bead-based activation and IL-2/IL-15-mediated expansion were used. This protocol can be applied for every possible CAR construct after being successfully transfected in HEK-293-based packaging cell lines.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"191 ","pages":"329-352"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143008161","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Evaluation of lymphocyte infiltration into cancer spheroids by immunofluorescent staining and 3D imaging. 免疫荧光染色和三维成像评价淋巴细胞浸润癌球体。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-14 DOI: 10.1016/bs.mcb.2024.10.015

Mireia Cruz De Los Santos, Andreas Lundqvist

引用次数: 0

Stereotactic injection of murine brain tumor cells for neuro-oncology studies. 立体定向注射小鼠脑肿瘤细胞用于神经肿瘤学研究。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-08-12 DOI: 10.1016/bs.mcb.2024.07.005

Camille Daviaud, María Cecilia Lira, Claire Vanpouille-Box, Mara De Martino

引用次数: 0

Measuring interaction force between T lymphocytes and their target cells using live microscopy and laminar shear flow chambers. 利用活体显微镜和层流剪切室测量T淋巴细胞与靶细胞之间的相互作用力。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-08 DOI: 10.1016/bs.mcb.2024.09.001

Sophie Goyard, Amandine Schneider, Jerko Ljubetic, Nicolas Inacio, Marie Juzans, Céline Cuche, Pascal Bochet, Vincenzo Di Bartolo, Andrés Alcover, Thierry Rose

{"title":"Measuring interaction force between T lymphocytes and their target cells using live microscopy and laminar shear flow chambers.","authors":"Sophie Goyard, Amandine Schneider, Jerko Ljubetic, Nicolas Inacio, Marie Juzans, Céline Cuche, Pascal Bochet, Vincenzo Di Bartolo, Andrés Alcover, Thierry Rose","doi":"10.1016/bs.mcb.2024.09.001","DOIUrl":"10.1016/bs.mcb.2024.09.001","url":null,"abstract":"Understanding the immunological synapse formation and dynamics can be enriched by measuring cell-cell interaction forces and their kinetics. Microscopy imaging reveals structural organization of the synapse, while physical methods detail its mechanical construction. Various techniques have been reported for measuring forces needed to rupture the interface between a T lymphocyte and its target cell but most of them measure one pair at a time. We describe here a laminar shear flow-based method that exerts dragging forces on T cell-target cells pairs immobilized on the surface of a flow chamber. Increasing flow rate allows us to observe the detachment of hundreds of cell conjugates on the wide field of a light transmission microscope. Monitoring precisely the flow rate gradient exerted on T cells readily yields synapse rupture measurements. Dragging forces are measured at the point of rupture as a linear function of the flow speed in minutes from 10pN to 20nN for each cell pair among a statistically representative cell population in the whole field of view of a single experiment. The output cells can be collected in multi-well plate sorted in the increasing order of rupture forces. We used this approach to unveil the involvement of the cytoskeleton regulator adenomatous polyposis coli (APC) in the stability of immunological synapses formed between human cytotoxic T cell and tumor target cells. APC is a polarity regulator and tumor suppressor associated with familial adenomatous polyposis and colorectal cancer. Reduced APC expression impairs T cell adhesion with tumor target cells suggesting an impact of APC mutation in anti-tumor immune defense.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"193 ","pages":"175-200"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143370861","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Expansion and activation of NK cells supported by accessory cells. Phenotypic and functional characterization. 辅助细胞支持NK细胞的扩增和活化。表型和功能特征。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2025-03-04 DOI: 10.1016/bs.mcb.2025.02.003

Cecilia Pesini, Pilar M Lanuza, Julián Pardo, Diego Sánchez-Martínez, Ariel Ramírez-Labrada

{"title":"Expansion and activation of NK cells supported by accessory cells. Phenotypic and functional characterization.","authors":"Cecilia Pesini, Pilar M Lanuza, Julián Pardo, Diego Sánchez-Martínez, Ariel Ramírez-Labrada","doi":"10.1016/bs.mcb.2025.02.003","DOIUrl":"https://doi.org/10.1016/bs.mcb.2025.02.003","url":null,"abstract":"Natural Killer cells (NK) are cytotoxic lymphocytes from the innate immune system that recognize and eliminate virally infected and tumor cells. Accordingly, manipulation of NK cells has been the focus of several immunotherapy protocols aimed at eradicating cancer cells. Allogeneic NK cell therapy was initially described over two decades ago, emphasizing KIR-mismatch's importance in preventing NK cell inhibition and promoting cytotoxicity and tumor elimination without inducing graft-versus-host disease (GvHD). While unstimulated NK cells have shown limited antitumoral activity in adoptive cell therapy, various activation and expansion protocols have been proposed to enhance their cytotoxic potential. Activated and expanded allogeneic NK cells, especially with the rise of chimeric antigen receptor (CAR) therapies, have attracted significant attention from academic and commercial sectors. Protocols typically involve using cytokines and stimulatory cells, such as Epstein-Barr virus (EBV)-transformed lymphoblastoid B cell lines (LCLs) or K562 leukemic cells, before or after NK cell enrichment. Here we present two different standardized protocols for NK cell activation and expansion, offering insights into NK cell-based immunotherapies for cancer treatment. We also present a comprehensive methodology for assessing NK cell-mediated cytotoxicity against Neuroblastoma cell lines in both 2D and 3D cultures. The comprehensive methodology presented here lays the foundation for further research in the field, driving advancements in NK cell-based therapies against malignancies.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"196 ","pages":"237-250"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144667968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Guidelines for the assessment of high endothelial venule functionality and health. 高内皮小静脉功能和健康评估指南。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2025-03-17 DOI: 10.1016/bs.mcb.2025.02.001

Kathryn A Jacobs, Gabriele Bergers

引用次数: 0