Methods in cell biology最新文献_第5页

In vitro re-challenge of CAR T cells. CAR T 细胞的体外再挑战。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-09-15 DOI: 10.1016/bs.mcb.2023.06.003

Clara Helena Klee, Alicia Villatoro, Nicholas Paul Casey, Else Marit Inderberg, Sébastien Wälchli

引用次数: 0

CD4+ T cells proliferation assay to analyze Mo-MDSCs suppressive function. 通过 CD4+ T 细胞增殖试验来分析 Mo-MDSCs 的抑制功能。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-10-24 DOI: 10.1016/bs.mcb.2023.04.005

María Iglesias-Escudero, David San Segundo, Marcos López-Hoyos

引用次数: 0

Estimating nitric oxide (NO) from MDSCs by Griess method. 用 Griess 法估算 MDSCs 中的一氧化氮（NO）。

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Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-10-11 DOI: 10.1016/bs.mcb.2023.07.004

Pronabesh Ghosh, Saikat Mukherjee, Soubhik Ghosh, Anwesha Gangopadhyay, Tarun Keswani, Anirban Sengupta, Samrat Sarkar, Arindam Bhattacharyya

{"title":"Estimating nitric oxide (NO) from MDSCs by Griess method.","authors":"Pronabesh Ghosh, Saikat Mukherjee, Soubhik Ghosh, Anwesha Gangopadhyay, Tarun Keswani, Anirban Sengupta, Samrat Sarkar, Arindam Bhattacharyya","doi":"10.1016/bs.mcb.2023.07.004","DOIUrl":"10.1016/bs.mcb.2023.07.004","url":null,"abstract":"The functional importance of nitric oxide (NO) in the fields of immunology concerning its antimicrobial, anti-tumoral, anti-inflammatory, and immunosuppressive effects have made it inevitable to study its secretion from various cells. Nitrogen oxide synthase (NOS) is the enzyme responsible for synthesizing NO and its three isoforms function in a cell-dependent manner. NO is oxidized rapidly to Reactive nitrogen oxide species (RNOS) through which the roles of NO are being carried out. One of the major immune cells secreting NO is myeloid-derived suppressor cells (MDSCs). The function of these MDSCs in the suppression of T-cell proliferation as well as T-cell differentiation is found to be dependent on NO secretion. Apart from T-cell suppressive activity, NO is also known to interfere with natural killer (NK) cell functions. A convenient method to estimate NO secretion is by using Griess reagent named after Johann Peter Griess. In this method, NO reacts with the reagents to form a colored azo dye detectable using a microplate reader at a wavelength of 548nm. In this chapter, we summarized the detailed method of estimating NO from MDSCs by the Griess method.","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":"140329973","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

Peritoneal carcinomatosis in mouse models. 小鼠腹膜癌模型。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-05 DOI: 10.1016/bs.mcb.2024.02.005

Leire Arrizabalaga, Claudia Augusta Di Trani, Aline Risson, Virginia Belsúe, Celia Gomar, Nuria Ardaiz, Pedro Berrondo, Fernando Aranda, Ángela Bella

{"title":"Peritoneal carcinomatosis in mouse models.","authors":"Leire Arrizabalaga, Claudia Augusta Di Trani, Aline Risson, Virginia Belsúe, Celia Gomar, Nuria Ardaiz, Pedro Berrondo, Fernando Aranda, Ángela Bella","doi":"10.1016/bs.mcb.2024.02.005","DOIUrl":"10.1016/bs.mcb.2024.02.005","url":null,"abstract":"Peritoneal carcinomatosis (PCa) represents a metastatic stage of a disease with unmet therapeutic options. Malignant cells from primary tumors (gastrointestinal or gynecologic malignancies) invade the peritoneal cavity and eventually seed onto peritoneal surfaces, with the omentum being the most common nest area. With a median survival of less than 6 months, PCa has a dismal prognosis that can be improved with treatments only available to a select few individuals with low tumor burden. Thus, the discovery of novel and effective therapies for this disease depends on reliable animal models. Here, we describe a method to generate syngeneic PCa mouse models based on intraperitoneal (i.p.) administration of tumor cells. This model allows to follow-up cancer progression in PCa models from ovarian and colorectal origins monitoring mice bodyweight changes, ascites development and overall survival. Moreover, luciferase-expressing tumor cells can also be used to assess tumor growth after i.p. injection through in vivo bioluminescence quantification. The establishment of reliable, easy-to-monitor and reproducible intraperitoneal syngeneic tumors models, as described here, is the first step to develop cutting-edge therapies against PCa.","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":"140331808","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

Preface. 序言

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Methods in cell biology Pub Date : 2024-01-01 DOI: 10.1016/S0091-679X(24)00112-2

José Manuel Bravo-San Pedro, Fernando Aranda, Aitziber Buqué, Lorenzo Galluzzi

引用次数: 0

Analysis of immunohistomorphological changes in the colonic mucosa in a high-saturated fat and high-cholesterol fed streptozotocin/nicotinamide diabetic rat model. 高饱和脂肪和高胆固醇喂养链脲佐菌素/烟酰胺糖尿病大鼠模型结肠粘膜免疫组织形态学变化分析。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-22 DOI: 10.1016/bs.mcb.2024.02.010

Marina Hernández-Martín, Aránzazu Bocanegra, Alba Garcimartín, Jousef Ángel Issa, Rocío Redondo-Castillejo, Adrián Macho-González, Juana Benedí, Francisco José Sánchez Muniz, María Elvira López-Oliva

{"title":"Analysis of immunohistomorphological changes in the colonic mucosa in a high-saturated fat and high-cholesterol fed streptozotocin/nicotinamide diabetic rat model.","authors":"Marina Hernández-Martín, Aránzazu Bocanegra, Alba Garcimartín, Jousef Ángel Issa, Rocío Redondo-Castillejo, Adrián Macho-González, Juana Benedí, Francisco José Sánchez Muniz, María Elvira López-Oliva","doi":"10.1016/bs.mcb.2024.02.010","DOIUrl":"10.1016/bs.mcb.2024.02.010","url":null,"abstract":"The mucosal surface of gastrointestinal tract is lined with epithelial cells that establish an effective barrier between the lumen and internal environment through intercellular junctions, preventing the passage of potentially harmful substances. The \"intestinal barrier function\" consist of a defensive system that prevent the passage of antigens, toxins, and microbial products, while maintains the correct development of the epithelial barrier, the immune system and the acquisition of tolerance toward dietary antigens and intestinal microbiota. Intestinal morphology changes subsequent to nutritional variations, stress, aging or diseases, which can also affect the composition of the microbiota, altering the homeostasis of the intestine. A growing body of evidence suggests that alterations in intestinal barrier function favor the development of exaggerated immune responses, leading to metabolic endotoxemia, which seems to be the origin of many chronic metabolic diseases such as type 2 diabetes mellitus (T2DM). Although the mechanisms are still unknown, the interaction between dietary patterns, gut microbiota, intestinal mucosa, and metabolic inflammation seems to be a key factor for the development of T2DM, among other diseases. This chapter details the different techniques that allow evaluating the morphological and molecular alterations that lead of the intestinal barrier dysfunction in a T2DM experimental model. To induce both diabetic metabolic disturbances and gut barrier disruption, Wistar rats were fed a high-saturated fat and high-cholesterol diet and received a single dose of streptozotocin/nicotinamide. This animal model may contribute to clarify the understanding of the role of intestinal barrier dysfunction on the late-stage T2DM etiology.","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":"140331856","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

Cloning and expansion of repetitive DNA sequences. 克隆和扩增重复 DNA 序列。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2022-11-28 DOI: 10.1016/bs.mcb.2022.10.013

Sophie L Williams, Gideon Coster

引用次数: 0

Generation of aneuploid cells and assessment of their ability to survive in presence of chemotherapeutic agents. 生成非整倍体细胞并评估其在化疗药物作用下的存活能力。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2022-11-24 DOI: 10.1016/bs.mcb.2022.10.012

Marica Rosaria Ippolito, Stefano Santaguida

{"title":"Generation of aneuploid cells and assessment of their ability to survive in presence of chemotherapeutic agents.","authors":"Marica Rosaria Ippolito, Stefano Santaguida","doi":"10.1016/bs.mcb.2022.10.012","DOIUrl":"10.1016/bs.mcb.2022.10.012","url":null,"abstract":"Aneuploidy is a condition in which cells have an abnormal number of chromosomes that is not a multiple of the haploid complement. It is known that aneuploidy has detrimental consequences on cell physiology, such as genome instability, metabolic and proteotoxic stress and decreased cellular fitness. Importantly, aneuploidy is a hallmark of tumors and it is associated with resistance to chemotherapeutic agents and poor clinical outcome. To shed light into how aneuploidy contributes to chemoresistance, we induced chromosome mis-segregation in human cancer cell lines, then treated them with several chemotherapeutic agents and evaluated the emergence of chemoresistance. By doing so, we found that elevation of chromosome mis-segregation promotes resistance to chemotherapeutic agents through the expansion of aneuploid karyotypes and subsequent selection of specific aneuploidies essential for cellular viability under those stressful conditions. Here, we describe a method to generate aneuploid cell populations and to evaluate their resistance to anti-cancer agents. This protocol has been already successfully employed and can be further utilized to accelerate the exploration of the role of aneuploidy in chemoresistance.","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":"139741404","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

Nascent DNA sequencing and its diverse applications in genome integrity research. 新生 DNA 测序及其在基因组完整性研究中的多种应用。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-01-03 DOI: 10.1016/bs.mcb.2022.12.011

Jacob Paiano, André Nussenzweig

{"title":"Nascent DNA sequencing and its diverse applications in genome integrity research.","authors":"Jacob Paiano, André Nussenzweig","doi":"10.1016/bs.mcb.2022.12.011","DOIUrl":"10.1016/bs.mcb.2022.12.011","url":null,"abstract":"Multiple DNA repair pathways and biological responses to DNA damage have evolved to protect cells from various types of lesions to which they are subjected. Although DNA repair systems are mechanistically distinct, all process the damaged region and then insert new bases to fill the gap. In 1969, Robert Painter developed an assay called \"unscheduled\" DNA synthesis (UDS), which measures DNA repair synthesis as the uptake of radiolabeled DNA precursors distinct from replicative synthesis. Contemporary detection of nascent DNA during repair by next-generation sequencing grants genome-wide information about the nature of lesions that threaten genome integrity. Recently, we developed the SAR-seq (synthesis associated with repair sequencing) method, which provides a high-resolution view of UDS. SAR-seq has been utilized to map programmed DNA repair sites in non-dividing neurons, replication initiation zones, monitor 53BP1 function in countering end-resection, and to identify regions of the genome that fail to complete replication during S phase but utilize repair synthesis during mitosis (MiDAS). As an example of SAR-seq, we present data showing that sites replicated during mitosis correspond to common fragile sites, which have been linked to tumor progression, cellular senescence, and aging.","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":"139741410","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

Using an ImageJ-based script to detect replication stress and associated cell cycle exit from G2 phase by fluorescence microscopy. 使用基于 ImageJ 的脚本，通过荧光显微镜检测复制应激和相关的细胞周期从 G2 期退出。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-03-25 DOI: 10.1016/bs.mcb.2023.02.003

Arne Lindqvist, Zhiyu Hao, Karen Akopyan

引用次数: 0