Methods in cell biology最新文献_第9页

Detection of myeloid-derived suppressor cells by flow cytometry. 通过流式细胞仪检测髓源性抑制细胞。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-10-11 DOI: 10.1016/bs.mcb.2023.06.006

Tianju Liu, Alyssa Rosek, Francina Gonzalez De Los Santos, Sem H Phan

{"title":"Detection of myeloid-derived suppressor cells by flow cytometry.","authors":"Tianju Liu, Alyssa Rosek, Francina Gonzalez De Los Santos, Sem H Phan","doi":"10.1016/bs.mcb.2023.06.006","DOIUrl":"10.1016/bs.mcb.2023.06.006","url":null,"abstract":"Recently discovered heterogeneous myeloid-derived suppressor cells (MDSCs) are some of the most discussed immunosuppressive cells in contemporary immunology, especially in the tumor microenvironment, and are defined primarily by their T cell immunosuppressive function. The importance of these cells extend to other chronic pathological conditions as well, including chronic infection, inflammation, and tissue remodeling. In many of these conditions, their accumulation/expansion correlates with disease progression, poor prognosis, and reduced survival, which highlights the potential of how these cells may be used in a clinical setting as both prognostic factor and therapeutic target. In healthy individuals, these cells are usually not present in the circulation. Therefore, monitoring this cell population is of potential clinical significance, and utility in basic research. However, these cells have a complex phenotype without one single marker of sufficient specificity for their identification. Flow cytometry is a powerful tool allowing multi-parameter analysis of heterogeneous cell populations, which makes it ideally suitable for the complex phenotypic analysis essential for identification and enumeration of circulating MDSCs. This approach has the potential to provide a novel clinically useful tool for assessment of prognosis and treatment outcomes. The protocol in this chapter describes a flow cytometric analysis to identify and quantify MDSCs from human or mouse whole blood leukocytes and peripheral blood mononuclear cells, as well as a single cell suspension from solid tissue, by using multicolor fluorescence-conjugated antibodies against their surface markers.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140329972","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

In vitro generation of murine myeloid-derived suppressor cells from hematopoietic progenitor cells. 从造血祖细胞体外生成小鼠髓源性抑制细胞。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-12-21 DOI: 10.1016/bs.mcb.2023.10.001

Lorraine M Hernandez-Delgado, Andrea Laboy-Figueroa, Alondra Lopez-Velazquez, Carolina Flores-Ortiz, Kevin Alicea-Torres

引用次数: 0

Single clonal tracking on biomimetic microtextured platforms for real-time guided migration analysis of myeloid-derived suppressor cell dissemination characteristics ex vivo. 在生物仿真微纹理平台上进行单克隆跟踪，对髓源性抑制细胞的体内外扩散特征进行实时引导迁移分析。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-02-21 DOI: 10.1016/bs.mcb.2024.01.002

Ana Panic, Jordan Moore, Daniel Gallego-Perez

{"title":"Single clonal tracking on biomimetic microtextured platforms for real-time guided migration analysis of myeloid-derived suppressor cell dissemination characteristics ex vivo.","authors":"Ana Panic, Jordan Moore, Daniel Gallego-Perez","doi":"10.1016/bs.mcb.2024.01.002","DOIUrl":"10.1016/bs.mcb.2024.01.002","url":null,"abstract":"Current strategies to undermine the deleterious influence of myeloid-derived suppressor cells (MDSCs) in the tumor microenvironment (TME) are lacking effective clinical solutions, in large part, due to insufficient knowledge on susceptible cellular and molecular targets. We describe here the application of biomimetic microfabricated platforms designed to analyze migratory phenotypes of MDSCs in the tumor niche ex vivo, which may enable accelerated therapeutic discovery. By mimicking the guided structural cues present in the physiological architecture of the TME, aligned microtopography substrates can elucidate potential interventions on migratory phenotypes of MDSCs at the single clonal level. Coupled with cellular and molecular biology analysis tools, our approach employs real-time tracking analysis of cell motility to probe the dissemination characteristics of MDSCs under guided migration conditions. These methods allow us to identify cellular subpopulations of interest based on their disseminative and suppressive capabilities. By doing so, we illustrate the potential of applying microscale engineering tools, in concert with dynamic live cell imaging and bioanalysis methods to uncover novel exploitable motility targets for advancing cancer therapy discovery. The inherent simplicity and extended application to a variety of contexts in tumor-associated cell migration render this method widely accessible to existing biological laboratory conditions and interests.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140329980","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

Radiotherapy protocols for mouse cancer model. 小鼠癌症模型的放射治疗方案。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-23 DOI: 10.1016/bs.mcb.2024.02.007

Eneko Garate-Soraluze, Javier Marco-Sanz, Irantzu Serrano-Mendioroz, Lucía Marrodán, Leticia Fernandez-Rubio, Sara Labiano, María E Rodríguez-Ruiz

{"title":"Radiotherapy protocols for mouse cancer model.","authors":"Eneko Garate-Soraluze, Javier Marco-Sanz, Irantzu Serrano-Mendioroz, Lucía Marrodán, Leticia Fernandez-Rubio, Sara Labiano, María E Rodríguez-Ruiz","doi":"10.1016/bs.mcb.2024.02.007","DOIUrl":"10.1016/bs.mcb.2024.02.007","url":null,"abstract":"Radiotherapy is a crucial treatment modality for cancer patients, with approximately 60% of individuals undergoing ionizing radiation as part of their disease management. In recent years, there has been a growing trend toward minimizing irradiation fields through the use of image-guided dosimetry and innovative technologies. These advancements allow for selective irradiation, delivering higher local doses while reducing the number of treatment sessions. Consequently, computer-assisted methods have significantly enhanced the effectiveness of radiotherapy in the curative and palliative treatment of various cancers. Although radiation therapy alone can effectively achieve local control in some cancer types, it may not be sufficient for others. As a result, further preclinical research is necessary to explore novel approaches including new schedules of radiotherapy treatments. Unfortunately, there is a concerning lack of correlation between clinical outcomes and experiments conducted on mouse models. We hypothesize that this disparity arises from the differences in irradiation strategies employed in preclinical studies compared to those used in clinical practice, which ultimately affects the translatability of findings to patients. In this study, we present two comprehensive radiotherapy protocols for the treatment of orthotopic melanoma and glioblastoma tumors. These protocols utilize a small animal radiation research platform, which is an ideal radiation device for delivering localized and precise X-ray doses to the tumor mass. By employing these platforms, we aim to limit the side effects associated with irradiating healthy surrounding tissues. Our detailed protocols offer a valuable framework for conducting preclinical studies that closely mimic clinical radiotherapy techniques, bridging the gap between experimental results and patient outcomes.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140331810","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

Immunophenotyping challenging tissue types using high-dimensional full spectrum flow cytometry. 利用高维全谱流式细胞仪对具有挑战性的组织类型进行免疫分型。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-07 DOI: 10.1016/bs.mcb.2024.02.014

Laura Ferrer-Font, Olivia K Burn, Johannes U Mayer, Kylie M Price

{"title":"Immunophenotyping challenging tissue types using high-dimensional full spectrum flow cytometry.","authors":"Laura Ferrer-Font, Olivia K Burn, Johannes U Mayer, Kylie M Price","doi":"10.1016/bs.mcb.2024.02.014","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.02.014","url":null,"abstract":"Technological advancements in fluorescence flow cytometry and an ever-expanding understanding of the complexity of the immune system, have led to the development of large flow cytometry panels, reaching up to 40 markers at the single-cell level. Full spectrum flow cytometry, that measures the full emission range of all the fluorophores present in the panel instead of only the emission peaks is now routinely used in many laboratories internationally, and the demand for this technology is rapidly increasing. With the capacity to use larger and more complex staining panels, optimized protocols are required for the best panel design, panel validation and high-dimensional data analysis outcomes. In addition, for ex vivo experiments, tissue preparation methods for single-cell analysis should also be optimized to ensure that samples are of the highest quality and are truly representative of tissues in situ. Here we provide optimized step-by-step protocols for full spectrum flow cytometry panel design, tissue digestion and panel optimization to facilitate the analysis of challenging tissue types.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140859600","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

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-21 DOI: 10.1016/bs.mcb.2024.02.034

Christine Péchoux, Fabrice Antigny, Frédéric Perros

{"title":"A correlated light and electron microscopy approach to study the subcellular localization of phosphorylated vimentin in human lung tissue.","authors":"Christine Péchoux, Fabrice Antigny, Frédéric Perros","doi":"10.1016/bs.mcb.2024.02.034","DOIUrl":"10.1016/bs.mcb.2024.02.034","url":null,"abstract":"Correlative microscopy is an important approach for bridging the resolution gap between fluorescence light and electron microscopy. Here, we describe a fast and simple method for correlative immunofluorescence and immunogold labeling on the same section to elucidate the localization of phosphorylated vimentin (P-Vim), a robust feature of pulmonary vascular remodeling in cells of human lung small arteries. The lung is a complex, soft and difficult tissue to prepare for transmission electron microscopy (TEM). Detailing the molecular composition of small pulmonary arteries (<500μm) would be of great significance for research and diagnostics. Using the classical methods of immunochemistry (either hydrophilic resin or thin cryosections), is difficult to locate small arteries for analysis by TEM. To address this problem and to observe the same structures by both light and electron microscopy, correlative microscopy is a reliable approach. Immunofluorescence enables us to know the distribution of P-Vim in cells but does not provide ultrastructural detail on its localization. Labeled structures selected by fluorescence microscope can be identified and further analyzed by TEM at high resolution. With our method, the morphology of the arteries is well preserved, enabling the localization of P-Vim inside pulmonary endothelial cells. By applying this approach, fluorescent signals can be directly correlated to the corresponding subcellular structures in areas of interest.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140855479","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

Analysis of the multiparametric cell cycle data. 多参数细胞周期数据分析。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-26 DOI: 10.1016/bs.mcb.2024.02.021

James W Jacobberger, R Michael Sramkoski, Tammy Stefan, Chris Bray, C Bruce Bagwell

引用次数: 0

Cellular senescence and aging at the crossroad between immunity and cancer. 免疫与癌症交叉路口的细胞衰老和老化。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 DOI: 10.1016/S0091-679X(24)00009-8

Oliver Kepp, Lorenzo Galluzzi, Giulia Petroni

引用次数: 0

Heat shock and thermotolerance in Caenorhabditis elegans: An overview of laboratory techniques. 草履虫的热休克和耐热性：实验室技术概述

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-12 DOI: 10.1016/bs.mcb.2024.02.001

Teresa Rubio-Tomás, Eva Alegre-Cortés, Eirini Lionaki, José M Fuentes, Nektarios Tavernarakis

引用次数: 0

Array tomography of in vivo labeled synaptic receptors. 体内标记突触受体的阵列断层成像。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-07 DOI: 10.1016/bs.mcb.2024.02.029

Sebastian Britz, Camilla Luccardini, Sebastian M Markert, Sean A Merrill, Jean-Louis Bessereau, Christian Stigloher

{"title":"Array tomography of in vivo labeled synaptic receptors.","authors":"Sebastian Britz, Camilla Luccardini, Sebastian M Markert, Sean A Merrill, Jean-Louis Bessereau, Christian Stigloher","doi":"10.1016/bs.mcb.2024.02.029","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.02.029","url":null,"abstract":"Array tomography (AT) allows one to localize sub-cellular components within the structural context of cells in 3D through the imaging of serial sections. Using this technique, the z-resolution can be improved physically by cutting ultra-thin sections. Nevertheless, conventional immunofluorescence staining of those sections is time consuming and requires relatively large amounts of costly antibody solutions. Moreover, epitopes are only readily accessible at the section's surface, leaving the volume of the serial sections unlabeled. Localization of receptors at neuronal synapses in 3D in their native cellular ultrastructural context is important for understanding signaling processes. Here, we present in vivo labeling of receptors via fluorophore-coupled tags in combination with super-resolution AT. We present two workflows where we label receptors at the plasma membrane: first, in vivo labeling via microinjection with a setup consisting of readily available components and self-manufactured microscope table equipment and second, live receptor labeling by using a cell-permeable tag. To take advantage of a near-to-native preservation of tissues for subsequent scanning electron microscopy (SEM), we also apply high-pressure freezing and freeze substitution. The advantages and disadvantages of our workflows are discussed.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140865741","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