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Protocol for screening potential targets for the treatment of hepatocellular carcinoma based on H22 tumor-bearing mouse model. 基于 H22 肿瘤小鼠模型的肝细胞癌潜在治疗靶点筛选方案。
IF 1.3
STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-10-17 DOI: 10.1016/j.xpro.2024.103193
Xiaoxiang Gao, Yinghui Qiu, Wei Liao, Chao Zhao
{"title":"Protocol for screening potential targets for the treatment of hepatocellular carcinoma based on H22 tumor-bearing mouse model.","authors":"Xiaoxiang Gao, Yinghui Qiu, Wei Liao, Chao Zhao","doi":"10.1016/j.xpro.2024.103193","DOIUrl":"10.1016/j.xpro.2024.103193","url":null,"abstract":"<p><p>Regulating cancer-related microRNAs (miRNAs) may become a new generation of therapeutic modalities for cancer treatment. Here, we describe a protocol based on hepatoma-22 (H22) tumor-bearing mice to screen potential targets for treating hepatocellular carcinoma (HCC). We detail the construction of H22 tumor-bearing mice and treatment with two natural compounds, Ulva lactuca L. polysaccharide (ULP) and 5-fluorouracil (5FU). We further describe the isolation of the tumor tissues for miRNA sequencing and the discovery and validation of potential miRNA gene targets against HCC. For complete details on the use and execution of this protocol, please refer to Qiu et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103193"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11532290/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142476492","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol to induce cell labeling in vivo and to trace labeled hematopoietic cells using mouse models. 利用小鼠模型诱导体内细胞标记并追踪标记的造血细胞。
IF 1.3
STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-10-21 DOI: 10.1016/j.xpro.2024.103408
Alina Syed, Michihiro Kobayashi, Momoko Yoshimoto
{"title":"Protocol to induce cell labeling in vivo and to trace labeled hematopoietic cells using mouse models.","authors":"Alina Syed, Michihiro Kobayashi, Momoko Yoshimoto","doi":"10.1016/j.xpro.2024.103408","DOIUrl":"10.1016/j.xpro.2024.103408","url":null,"abstract":"<p><p>A combination of hematopoietic stem cell (HSC)- and endothelial cell (EC)-lineage-tracing mouse models enables us to determine blood cell origins. We present a protocol to induce cell labeling in vivo and to trace labeled hematopoietic cells to segregate their origins. We describe the steps for harvesting various hematopoietic tissues, antibody staining, and analyzing the Tomato+ percentages within each immune cell population. We also show how to estimate HSC- and EC-derived percentages of the target cell populations. For complete details on the use and execution of this protocol, please refer to Kobayashi et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103408"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11532922/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142509490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A luminescent-based protocol for NAD+/NADH detection in C. elegans, mice, and human whole blood. 一种基于发光的方案，用于检测线虫、小鼠和人类全血中的 NAD+/NADH。
IF 1.3
STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-11-01 DOI: 10.1016/j.xpro.2024.103428
He-Ling Wang, Jianying Zhang, Shu-Qin Cao, Maria Jose Lagartos-Donate, Shi-Qi Zhang, Sofie Lautrup, Zeping Hu, Costas A Lyssiotis, Riekelt H Houtkooper, Evandro F Fang
{"title":"A luminescent-based protocol for NAD<sup>+</sup>/NADH detection in C. elegans, mice, and human whole blood.","authors":"He-Ling Wang, Jianying Zhang, Shu-Qin Cao, Maria Jose Lagartos-Donate, Shi-Qi Zhang, Sofie Lautrup, Zeping Hu, Costas A Lyssiotis, Riekelt H Houtkooper, Evandro F Fang","doi":"10.1016/j.xpro.2024.103428","DOIUrl":"10.1016/j.xpro.2024.103428","url":null,"abstract":"<p><p>Here, we present a NAD<sup>+</sup>/NADH detection assay for evaluating NAD<sup>+</sup>, NADH, and NAD<sup>+</sup>/NADH ratio across diverse biological models, including Caenorhabditis elegans, mouse muscle tissue, mouse whole blood, and human whole blood. We describe steps for sample collection and preparation from different models as well as detection and calculation of NAD<sup>+</sup> and NADH levels. This protocol is applicable for quantifying cellular/tissue NAD<sup>+</sup> and NADH levels across different biological models.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103428"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11567065/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142565106","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for muscle fiber type and cross-sectional area analysis in cryosections of whole lower mouse hindlimbs. 小鼠后肢整体冷冻切片的肌纤维类型和横截面积分析程序
IF 1.3
STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-11-01 DOI: 10.1016/j.xpro.2024.103424
Edmund Battey, Matthieu Dos Santos, Dipsikha Biswas, Pascal Maire, Kei Sakamoto
{"title":"Protocol for muscle fiber type and cross-sectional area analysis in cryosections of whole lower mouse hindlimbs.","authors":"Edmund Battey, Matthieu Dos Santos, Dipsikha Biswas, Pascal Maire, Kei Sakamoto","doi":"10.1016/j.xpro.2024.103424","DOIUrl":"10.1016/j.xpro.2024.103424","url":null,"abstract":"<p><p>We outline a protocol to visualize all mouse lower hindlimb skeletal muscles simultaneously. We describe procedures for orientating the whole lower hindlimb in gum tragacanth prior to freezing, simplifying the proceeding experimental steps, and enhancing the comprehensiveness of characterizations. We then detail steps for quantifying muscle fiber size and fiber type characteristics in a single cryosection using histochemistry and immunofluorescence. This protocol can be applied to histological and (immuno)histochemical evaluations such as muscle regeneration, fibrosis, enzymatic activity, and glycogen content.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103424"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11565383/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142565111","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol to phenotype and quantify mycobacteria-specific myeloid cells from human airways by mass cytometry. 利用质谱仪对人体气道中的分枝杆菌特异性髓系细胞进行表型和量化的方法。
IF 1.3
STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-12-13 DOI: 10.1016/j.xpro.2024.103463
Agano Kiravu, Virgine Rozot, Lauren Cruywagen, Andrea Gutschmidt, Nelita DuPlessis, Elisa Nemes
{"title":"Protocol to phenotype and quantify mycobacteria-specific myeloid cells from human airways by mass cytometry.","authors":"Agano Kiravu, Virgine Rozot, Lauren Cruywagen, Andrea Gutschmidt, Nelita DuPlessis, Elisa Nemes","doi":"10.1016/j.xpro.2024.103463","DOIUrl":"10.1016/j.xpro.2024.103463","url":null,"abstract":"<p><p>Alveolar macrophages and other myeloid cells in the human airways are the primary cell types responding to respiratory pathogens. Here, we present a protocol for in vitro stimulation of cryopreserved human bronchoalveolar lavage (BAL) cells with mycobacterial antigens for phenotyping and quantifying proinflammatory cytokine responses in myeloid cells by mass cytometry. We demonstrate that the measure of markers of myeloid lineage and function is stable after freezing stained cells thereby allowing for batched analyses and/or machine downtime.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103463"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11699420/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142824550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for the development of a bioluminescent AML-PDX mouse model for the evaluation of CAR T cell therapy. 开发用于评估 CAR T 细胞疗法的生物发光 AML-PDX 小鼠模型的方案。
IF 1.3
STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-12-12 DOI: 10.1016/j.xpro.2024.103522
Mireia Mayoral Safont, Calum Leitch, Mihaela Popa, May Eriksen Gjerstad, Benjamin Caulier, Else Marit Inderberg, Sébastien Wälchli, Pascal Gelebart, Emmet Mc Cormack
{"title":"Protocol for the development of a bioluminescent AML-PDX mouse model for the evaluation of CAR T cell therapy.","authors":"Mireia Mayoral Safont, Calum Leitch, Mihaela Popa, May Eriksen Gjerstad, Benjamin Caulier, Else Marit Inderberg, Sébastien Wälchli, Pascal Gelebart, Emmet Mc Cormack","doi":"10.1016/j.xpro.2024.103522","DOIUrl":"10.1016/j.xpro.2024.103522","url":null,"abstract":"<p><p>Patient-derived xenograft (PDX) models of acute myeloid leukemia (AML-PDX) offer advantages over cell line models by capturing the complexity and heterogeneity of patient-derived samples. Here, we present a protocol for developing a bioluminescent AML-PDX model in mice to evaluate chimeric antigen receptor (CAR) T cell therapy. We describe steps for transducing, engrafting, expanding, and enriching AML-PDX cells. We then detail procedures for in vitro and in vivo validation of the AML-PDX model for the evaluation of CAR T cell immunotherapy. For complete details on the use and execution of this protocol, please refer to Caulier et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103522"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11699402/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142824586","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for in vivo recording of neural activity in deep structures of mice brain via gradient lenses by calcium imaging.
IF 1.3
STAR Protocols Pub Date : 2024-12-20 DOI: 10.1016/j.xpro.2024.103534
Stanislav Cherepanov, Patrice Mollard, Agnes O Martin
{"title":"Protocol for in vivo recording of neural activity in deep structures of mice brain via gradient lenses by calcium imaging.","authors":"Stanislav Cherepanov, Patrice Mollard, Agnes O Martin","doi":"10.1016/j.xpro.2024.103534","DOIUrl":"https://doi.org/10.1016/j.xpro.2024.103534","url":null,"abstract":"<p><p>Calcium (Ca<sup>2+</sup>) imaging is a viable approach for imaging neuronal activity patterns in local brain circuits in living animals. Here, we present a protocol for gradient lens implantation in deep brain structures followed by in vivo Ca<sup>2+</sup> imaging. We describe in detail the steps for surgery preparation, followed by lens implantation, setup for awake head-fixed imaging, and the recording process. For complete details on the use and execution of this protocol, please refer to Cherepanov et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103534"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142872866","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol to culture Leishmania protozoans for lipophosphoglycan extraction and purification.
IF 1.3
STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-12-04 DOI: 10.1016/j.xpro.2024.103468
Lisa Ulrike Teufel, Jéssica Cristina Dos Santos
{"title":"Protocol to culture Leishmania protozoans for lipophosphoglycan extraction and purification.","authors":"Lisa Ulrike Teufel, Jéssica Cristina Dos Santos","doi":"10.1016/j.xpro.2024.103468","DOIUrl":"10.1016/j.xpro.2024.103468","url":null,"abstract":"<p><p>Lipophosphoglycan (LPG) is a macromolecule on the surface of Leishmania spp. parasites. The biochemical structure of LPG varies throughout the parasites' life cycle between proliferative (procyclic) and infective (metacyclic) stages, as well as between species and strains. Here, we outline a protocol for growing Leishmania parasites in vitro to harvest LPG. We describe steps for parasite differentiation and LPG extraction and purification. LPG has applications in medical research, such as in trained immunity and immunotherapy for cancer. For complete details on the use and execution of this protocol, please refer to dos Santos et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103468"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11664164/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142786790","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for immunofluorescence staining and large-scale analysis to quantify microglial cell morphology at single-cell resolution in mice.
IF 1.3
STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-12-04 DOI: 10.1016/j.xpro.2024.103467
Frida Lind-Holm Mogensen, Corrado Ameli, Alexander Skupin, Alessandro Michelucci
{"title":"Protocol for immunofluorescence staining and large-scale analysis to quantify microglial cell morphology at single-cell resolution in mice.","authors":"Frida Lind-Holm Mogensen, Corrado Ameli, Alexander Skupin, Alessandro Michelucci","doi":"10.1016/j.xpro.2024.103467","DOIUrl":"10.1016/j.xpro.2024.103467","url":null,"abstract":"<p><p>Here, we present a protocol for quantifying microglial cell morphology at the single-cell level in mice. We provide comprehensive details, starting from optimal mouse brain dissection to computational analyses of up to 350 microglial cells per brain slice. Analyzing the morphology of microglial cells is essential for understanding their functional and activation states in different conditions, including during disease development and progression, as well as for assessing the effect of therapeutic interventions. For complete details on the use and execution of this protocol, please refer to Lind-Holm Mogensen et al.<sup>1</sup> and Fixemer et al.<sup>2</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103467"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11653134/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142786809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol to measure glucose utilization in mouse tissues using radiolabeled 2-deoxy-D-glucose.
IF 1.3
STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-12-06 DOI: 10.1016/j.xpro.2024.103478
Cyun-Ming Wu, Hao-Yun Li, Wen-Yi Li, Chih-Neng Hsu, Wei-Shun Yang, Gwo-Tsang Chuang, Tony Pan-Hou Che, Tung-Yuan Lee, Hsiao-Lin Lee, Siow-Wey Hee, Jiin-Horng Lee, Daniel Liao, Karen Chia-Wen Liao, Ya-Wen Liu, Chin-Feng Hsuan, Yi-Cheng Chang, Shao-Lun Chu
{"title":"Protocol to measure glucose utilization in mouse tissues using radiolabeled 2-deoxy-D-glucose.","authors":"Cyun-Ming Wu, Hao-Yun Li, Wen-Yi Li, Chih-Neng Hsu, Wei-Shun Yang, Gwo-Tsang Chuang, Tony Pan-Hou Che, Tung-Yuan Lee, Hsiao-Lin Lee, Siow-Wey Hee, Jiin-Horng Lee, Daniel Liao, Karen Chia-Wen Liao, Ya-Wen Liu, Chin-Feng Hsuan, Yi-Cheng Chang, Shao-Lun Chu","doi":"10.1016/j.xpro.2024.103478","DOIUrl":"10.1016/j.xpro.2024.103478","url":null,"abstract":"<p><p>2-deoxy-D-glucose (2DG) is a glucose analog converted to 2-deoxy-D-glucose-6-phosphate (2DG-6P) by hexokinase in glycolysis. While 2DG commonly measures glucose uptake, 2DG-6P detects glucose utilization. Here, we present a protocol to measure glucose utilization in various tissues after entering a mouse's body using radiolabeled 2DG. We describe steps for preparing mice and chemicals, extracting blood, adding chemicals, and dissolving tissue. We then detail procedures for calculating glucose utilization using the trapezoid rule.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103478"},"PeriodicalIF":1.3,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11656071/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142792279","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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