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Protocol for isolation and culture establishment of adult murine neural stem cells using tissue grinder and dissociator-based mechanical dissociation. 使用组织研磨机和基于解离剂的机械解离分离和培养建立成年小鼠神经干细胞的方案。
IF 1.4
STAR Protocols Pub Date : 2026-10-20 DOI: 10.1016/j.xpro.2026.104908
Vanessa Arnold, Anna Wolfram, Anastassiya Moskalchuk, Vijayasarathy Sampath-Kumar, Abhirami Sunil, Sogand Ahari, Lisa Sevenich
{"title":"Protocol for isolation and culture establishment of adult murine neural stem cells using tissue grinder and dissociator-based mechanical dissociation.","authors":"Vanessa Arnold, Anna Wolfram, Anastassiya Moskalchuk, Vijayasarathy Sampath-Kumar, Abhirami Sunil, Sogand Ahari, Lisa Sevenich","doi":"10.1016/j.xpro.2026.104908","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104908","url":null,"abstract":"<p><p>Here, we present an enzyme-free workflow for isolating murine adult neural stem cells (aNSCs) using tissue grinder- and dissociator-based mechanical dissociation (TIGR-MD). We describe dissection of the dentate gyrus and subventricular zone, mechanical tissue dissociation, cell recovery, and establishment of primary neurosphere cultures. We further provide procedures for neurosphere maintenance, passaging, and validation. TIGR-MD avoids enzymatic exposure during initial tissue dissociation and supports standardized establishment of primary murine aNSC cultures.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104908"},"PeriodicalIF":1.4,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149228795","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
An improved protocol to isolate enriched immune cells from murine intestinal epithelium and lamina propria. 从小鼠肠上皮和固有层分离富集免疫细胞的改进方案。
IF 1.4
STAR Protocols Pub Date : 2026-10-20 DOI: 10.1016/j.xpro.2026.104864
Sydney B Crotts, Virginia Smith Shapiro
{"title":"An improved protocol to isolate enriched immune cells from murine intestinal epithelium and lamina propria.","authors":"Sydney B Crotts, Virginia Smith Shapiro","doi":"10.1016/j.xpro.2026.104864","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104864","url":null,"abstract":"<p><p>The intestine is home to more immune cells than any other non-lymphatic peripheral tissue, but research of these cells is stymied by the technical difficulties involved in isolating them. Here, we describe a protocol to isolate highly viable immune cells from murine intestinal epithelium and lamina propria. This protocol can be used to characterize cells by flow cytometry or single-cell transcriptomics, as in Crotts et al., or by other analysis methods.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104864"},"PeriodicalIF":1.4,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149228812","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 for simulating cell-contact dynamics of proliferating epithelia using the vertex model. 使用顶点模型模拟增殖上皮细胞接触动力学的方案。
IF 1.4
STAR Protocols Pub Date : 2026-10-19 DOI: 10.1016/j.xpro.2026.104909
Tanmoy Sarkar, Jeffrey J Lovelace, Matej Krajnc, Rosemary Mwithiga, Subramanian P Ramanathan
{"title":"Protocol for simulating cell-contact dynamics of proliferating epithelia using the vertex model.","authors":"Tanmoy Sarkar, Jeffrey J Lovelace, Matej Krajnc, Rosemary Mwithiga, Subramanian P Ramanathan","doi":"10.1016/j.xpro.2026.104909","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104909","url":null,"abstract":"<p><p>Vertex models simulate epithelial mechanics at single-cell resolution but represent cell division as a single in-plane split, agnostic to the differential apicobasal geometry of mitotic cells. Here, we present a protocol that simulates basal mitosis as two sequential changes: basal-cell diminution followed by daughter-cell reinsertion. Apical-side simulations can be performed with minimal code modification. We provide instructions for generating initial configurations, running simulations, and quantifying mechanical and topological rearrangements using command-line operations accessible to experimental biologists. For complete details on the use and execution of this protocol, please refer to Ramanathan et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104909"},"PeriodicalIF":1.4,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149228851","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
Image-guided focused ultrasound for non-invasive modulation of superficial and deep tissues. 图像引导聚焦超声对浅表和深部组织的无创调制。
IF 1.4
STAR Protocols Pub Date : 2026-10-19 DOI: 10.1016/j.xpro.2026.104900
Namrata Sangwan, N Chris Chaggares, Zahra Torbatian, Thomas Anthony Anderson
{"title":"Image-guided focused ultrasound for non-invasive modulation of superficial and deep tissues.","authors":"Namrata Sangwan, N Chris Chaggares, Zahra Torbatian, Thomas Anthony Anderson","doi":"10.1016/j.xpro.2026.104900","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104900","url":null,"abstract":"<p><p>Focused ultrasound (FUS) enables non-invasive, spatially precise modulation of biological tissues. This protocol describes assembly, calibration, and implementation of a diagnostic ultrasound-guided FUS (dUS-FUS) platform for real-time targeting of superficial and deep tissues in small-animal models. We demonstrate reversible sciatic nerve modulation in a rat acute pain model and assess sensory and motor functions using behavioral tests. The platform provides a reproducible workflow for image-guided tissue modulation and can be adapted to diverse preclinical applications. For complete details on the use and execution of this protocol, please refer to Sangwan et al.<sup>1</sup><sup>,</sup><sup>2</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104900"},"PeriodicalIF":1.4,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149224946","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 for integrated phosphoproteomic and proteomic analysis of the mouse ventral tegmental area with functional and kinase motif enrichment. 具有功能和激酶基序富集的小鼠腹侧被盖区综合磷蛋白质组学和蛋白质组学分析方案。
IF 1.4
STAR Protocols Pub Date : 2026-10-17 DOI: 10.1016/j.xpro.2026.104911
Cody C Diezel, Paul R Langlais, Todd W Vanderah, Arthur C Riegel
{"title":"Protocol for integrated phosphoproteomic and proteomic analysis of the mouse ventral tegmental area with functional and kinase motif enrichment.","authors":"Cody C Diezel, Paul R Langlais, Todd W Vanderah, Arthur C Riegel","doi":"10.1016/j.xpro.2026.104911","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104911","url":null,"abstract":"<p><p>We present a protocol for label-free proteomic and phosphoproteomic analysis of the mouse ventral tegmental area (VTA). We describe steps for combining region-specific biopsy-punch collection, parallel proteomic and phosphoproteomic processing, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and downstream enrichment analysis. We detail procedures for SynGO-based synaptic localization, g:Profiler/DAVID pathway enrichment, and iceLogo/pLogo motif analysis. This protocol enables quantitative assessment of protein abundance, phosphorylation dynamics, and functional interpretation from limited brain region samples. For complete details on the use and execution of this protocol, please refer to Diezel et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104911"},"PeriodicalIF":1.4,"publicationDate":"2026-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149217015","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 for eVLP-mediated CRISPR-Cas9 and prime editing in pancreatic cancer cell lines. 胰腺癌细胞系中evlp介导的CRISPR-Cas9和引物编辑方法
IF 1.4
STAR Protocols Pub Date : 2026-10-17 DOI: 10.1016/j.xpro.2026.104904
Chloe Timmins-Young, Marco Cordani, Elisa Giovannetti, Despoina Aslanoglou, Alvaro Gutierrez, Eithne Costello, Mark Glenn
{"title":"Protocol for eVLP-mediated CRISPR-Cas9 and prime editing in pancreatic cancer cell lines.","authors":"Chloe Timmins-Young, Marco Cordani, Elisa Giovannetti, Despoina Aslanoglou, Alvaro Gutierrez, Eithne Costello, Mark Glenn","doi":"10.1016/j.xpro.2026.104904","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104904","url":null,"abstract":"<p><p>Pancreatic ductal adenocarcinoma (PDAC) cell lines resist transfection, limiting gene-editing experiments. Here, we present a protocol for delivering CRISPR-Cas9 or prime editors into PDAC cell lines using retrovirus-derived engineered virus-like particles (eVLPs). We describe steps for generating eVLPs, packaging CRISPR-Cas9 ribonucleoproteins or prime editors with engineered prime-editing guide RNAs (epegRNAs), and transducing human PDAC cells. We use the HEK3 locus and NQO1 (NAD(P)H quinone dehydrogenase 1) exon 6 as example edits and controls, respectively. This protocol adapts readily to other genomic sites.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104904"},"PeriodicalIF":1.4,"publicationDate":"2026-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149216977","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 for the isolation, enrichment, and culture of murine adipose progenitor cells. 小鼠脂肪祖细胞的分离、富集和培养方案。
IF 1.4
STAR Protocols Pub Date : 2026-10-16 DOI: 10.1016/j.xpro.2026.104901
Ghazaleh Dadashizadeh, Fadi Khalaf, Stephanie Wojtowicz-Piotrowski, Marc G Jeschke
{"title":"Protocol for the isolation, enrichment, and culture of murine adipose progenitor cells.","authors":"Ghazaleh Dadashizadeh, Fadi Khalaf, Stephanie Wojtowicz-Piotrowski, Marc G Jeschke","doi":"10.1016/j.xpro.2026.104901","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104901","url":null,"abstract":"<p><p>We present a protocol for isolating and culturing murine adipose progenitor cells (APCs) from adipose tissue. We describe steps for tissue dissection, unit-normalized enzymatic digestion, stromal vascular fraction (SVF) isolation, APC enrichment, and cell culture. We then detail quality control procedures that maximize cell yield, viability, and purity. This protocol combines enzymatic SVF isolation with magnetic-activated cell sorting enrichment to obtain a highly enriched APC population. It emphasizes reproducibility through unit-normalized digestion, controlled processing conditions, and standardized quality control metrics.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104901"},"PeriodicalIF":1.4,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149207949","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
STAR Protocols question-and-answer series: Flow cytometry. STAR协议问答系列：流式细胞术。
IF 1.4
STAR Protocols Pub Date : 2026-10-15 DOI: 10.1016/j.xpro.2026.104895
Jeremy Petravicz
{"title":"STAR Protocols question-and-answer series: Flow cytometry.","authors":"Jeremy Petravicz","doi":"10.1016/j.xpro.2026.104895","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104895","url":null,"abstract":"<p><p>Flow cytometry is an integral part of the research scientist's toolbox. However, it is a complex methodology, involving decisions from antibody panel design to gating strategies to obtain the best results. Because it is so commonly used, there are many frequently asked questions about the protocols underlying its use. In this Backstory, STAR Protocols authors, reviewers, and advisory board members share their knowledge and expertise regarding common technical questions about flow cytometry.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104895"},"PeriodicalIF":1.4,"publicationDate":"2026-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149197027","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
Collaboration in action: Protocol development across the MRC National Mouse Genetics Network. 行动中的合作：跨MRC国家小鼠遗传学网络的协议开发。
IF 1.4
STAR Protocols Pub Date : 2026-10-15 DOI: 10.1016/j.xpro.2026.104898
Debora Bogani, Linus F Reichenbach, Jillian L Barlow, Karen Blyth
{"title":"Collaboration in action: Protocol development across the MRC National Mouse Genetics Network.","authors":"Debora Bogani, Linus F Reichenbach, Jillian L Barlow, Karen Blyth","doi":"10.1016/j.xpro.2026.104898","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104898","url":null,"abstract":"<p><p>Collaboration and multidisciplinary approaches are key factors in the advancement of scientific research. This Backstory introduces the guiding philosophy and the collaborative nature of the MRC National Mouse Genetics Network (NMGN) and serves to set the foundation for the contents of this STAR Protocols focused collection. The collection brings together protocols and Primers that highlight methodological advances in mouse genetics, disease modelling, phenotyping, data integration, and related experimental approaches developed across the NMGN. Together, these contributions illustrate how distributed expertise, shared infrastructure, and coordinated protocol development can strengthen reproducibility, support responsible use of in vivo models and emerging alternatives, and accelerate translational biomedical research.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104898"},"PeriodicalIF":1.4,"publicationDate":"2026-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149196986","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 for quantifying mitotic progression in adherent and suspension cells by live-cell imaging. 通过活细胞成像定量贴壁细胞和悬浮细胞有丝分裂进程的方法。
IF 1.4
STAR Protocols Pub Date : 2026-10-13 DOI: 10.1016/j.xpro.2026.104899
Siamak A Kamranvar
{"title":"Protocol for quantifying mitotic progression in adherent and suspension cells by live-cell imaging.","authors":"Siamak A Kamranvar","doi":"10.1016/j.xpro.2026.104899","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104899","url":null,"abstract":"<p><p>Live-cell imaging is used to monitor mitotic progression in adherent cells; however, its application to suspension cells remains limited. Here, we present a protocol for quantifying mitotic progression in adherent and suspension cells by monitoring chromatin condensation using the live-cell DNA probe SPY-DNA. We describe steps for cell synchronization, cell plating, SPY-DNA labeling, and live-cell imaging. We then detail procedures for surface plot visualization and intensity profile analysis. This approach enables direct comparison of mitotic progression between adherent and suspension cells. For complete details on the use and execution of this protocol, please refer to Kamranvar et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104899"},"PeriodicalIF":1.4,"publicationDate":"2026-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149184821","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
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