Jove-Journal of Visualized Experiments最新文献_第3页

Isolation and Flow Cytometric Assessment of Neuroimmune Interactions in a Mini-Stroke Murine Model. 小中风小鼠模型中神经免疫相互作用的分离和流式细胞术评估。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-20 DOI: 10.3791/68308

Song Wang, Jingyi Yao, Yitong Du, Song Zhang, Jiangxu Wu, Sichen Wang, Yuhan Sun, Dan Xie

{"title":"Isolation and Flow Cytometric Assessment of Neuroimmune Interactions in a Mini-Stroke Murine Model.","authors":"Song Wang, Jingyi Yao, Yitong Du, Song Zhang, Jiangxu Wu, Sichen Wang, Yuhan Sun, Dan Xie","doi":"10.3791/68308","DOIUrl":"https://doi.org/10.3791/68308","url":null,"abstract":"Stroke is a leading cause of death and long-term disability worldwide, with ischemic stroke representing the majority of cases. Following an ischemic stroke, resident and infiltrating immune cells become activated, contributing to further neuronal damage. However, the roles of the immune system in the pathology of ischemic stroke are not fully understood, largely due to the complex and dynamic regulation of immune responses in reaction to changes in the microenvironment during neuroinflammation. Therefore, it is essential to monitor and analyze the activation of resident and infiltrating immune cells over time after an ischemic stroke. In this study, we present a protocol for assessing the function of these immune cells following ischemic stroke using flow cytometric analysis in a mini-stroke model. We microdissect the infarcted brain tissue at specific time points and then dissociate it into a single-cell suspension using both mechanical and enzymatic methods. The cells are passed through a 70 µm cell strainer and labeled with fluorescently tagged antibody cocktails before being quantified by flow cytometric analysis. While this assay was specifically developed to investigate neuroimmune interactions after ischemic stroke, it can also be easily adapted to study neuroimmune mechanisms in other neurodegenerative diseases, such as multiple sclerosis, Parkinson's disease, and Alzheimer's disease.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Blue Light-inducible Semi-random Mutagenesis on Specific Exogenous DNA in Escherichia coli BL21 Strain. 蓝光诱导的大肠杆菌BL21株外源DNA的半随机突变

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-20 DOI: 10.3791/68283

Haitao Zhou, Wen Lin, Xiangyu Ma, Zixin Huang, Ziming Tian, Tianyue Dai, Xinran Ma, Ruoqin Hao, Duojia Li, Ziyi Wang

{"title":"Blue Light-inducible Semi-random Mutagenesis on Specific Exogenous DNA in Escherichia coli BL21 Strain.","authors":"Haitao Zhou, Wen Lin, Xiangyu Ma, Zixin Huang, Ziming Tian, Tianyue Dai, Xinran Ma, Ruoqin Hao, Duojia Li, Ziyi Wang","doi":"10.3791/68283","DOIUrl":"https://doi.org/10.3791/68283","url":null,"abstract":"Mutagenesis technologies have been widely used in variation breeding, directed evolution, and protein engineering. However, most methods can only induce random genomic DNA mutations that are uncontrollable, leading to deformities or even lethal effects. Here, by leveraging the fusion of three systems: cytosine base editor (CBE), blue light-inducible p-Mag/n-Mag elements, and split T7 RNA polymerase, this outlined method incorporates a blue-light-controllable, DNA region-specific, semi-random mutagenesis system. This system can mutate cytosine to thymine on DNAs starting from the T7 promoter and ending in the T7 terminator. Any exogenous gene downstream of the T7 promoter can be potentially mutated. These tools were also designed in two parts to make the system blue-light-controllable. The first part was created by fusing CBE with n-Mag and the N-terminal-T7 RNA polymerase gene. The second part involved fusing p-Mag with the C-terminal-T7 RNA polymerase. Under blue light, these two parts were assembled to function as a mutation generator on DNAs between the T7 promoter and the T7 terminator. Without blue light, p-Mag is detached from n-Mag, releasing the CBE from editing the DNA. The aim of this protocol is to generate region-specific random mutations in Escherichia coli with blue light control and further validate the mutated plasmids. The system can be adapted to an in vivo evolution system where the function of the gene of interest can be selected, and the desired mutations can be collected for further study.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Correlative Optical Spectroscopy and Mass Spectrometry Imaging Methodology to Visualise Drug Distribution in a Soft Tissue Section. 相关光谱学和质谱成像方法在软组织切片中可视化药物分布。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-20 DOI: 10.3791/67383

Vasundhara Tyagi, Alex Dexter, Jean-Luc Vorng, Dimitrios Tsikritsis, Ian S Gilmore, Richard H Guy, Mila Boncheva Bettex, Natalie A Belsey

{"title":"Correlative Optical Spectroscopy and Mass Spectrometry Imaging Methodology to Visualise Drug Distribution in a Soft Tissue Section.","authors":"Vasundhara Tyagi, Alex Dexter, Jean-Luc Vorng, Dimitrios Tsikritsis, Ian S Gilmore, Richard H Guy, Mila Boncheva Bettex, Natalie A Belsey","doi":"10.3791/67383","DOIUrl":"https://doi.org/10.3791/67383","url":null,"abstract":"A new correlative imaging methodology combining sub-micron spatial resolution with sensitive drug detection by optical spectroscopy and mass spectrometry imaging has been developed to visualise chemical distribution upon drug application in soft tissue. In this example, the method has been tested on excised skin tissue after in vitro topical application of a commercial nonsteroidal anti-inflammatory drug product for 16 hours. Non-destructive optical spectroscopic methods, including stimulated Raman scattering, second harmonic generation, and two-photon fluorescence microscopies, were first employed to map the skin structure and morphology. Subsequently, the same skin tissue samples were analysed via time-of-flight secondary ion mass spectrometry (ToF-SIMS), which enabled enhanced sensitivity for the detection of diclofenac across the outermost skin layers - epidermis and dermis. Image registration methods were devised to integrate the optical and mass spectrometric data. This approach combines label-free, high-resolution visualisation of tissue structure with sensitive chemical detection, and represents a valuable tool for investigating drug distribution in skin and potentially, in other soft biological tissues.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585334","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Profiling of Permethylated Mucin O-glycans Using Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry. 基质辅助激光解吸/电离飞行时间质谱分析过甲基化粘蛋白o -聚糖

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-20 DOI: 10.3791/67751

Dimitrios Latousakis, Nathalie Juge

引用次数: 0

RNA-Seq Reveals Th17 Cell Differentiation Pathway as a Mechanism of Radiation-Induced Brain Injury. RNA-Seq揭示Th17细胞分化途径在辐射性脑损伤中的作用机制

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-20 DOI: 10.3791/68322

Manqiang Sun, Bichun Zhao, Chao Wang, Shunyao Ma, Zejin Shen, Jiafei Xi, Junnian Zhou, Yali Jia, Wen Yue

{"title":"RNA-Seq Reveals Th17 Cell Differentiation Pathway as a Mechanism of Radiation-Induced Brain Injury.","authors":"Manqiang Sun, Bichun Zhao, Chao Wang, Shunyao Ma, Zejin Shen, Jiafei Xi, Junnian Zhou, Yali Jia, Wen Yue","doi":"10.3791/68322","DOIUrl":"https://doi.org/10.3791/68322","url":null,"abstract":"Radiotherapy is a prevalent therapeutic modality for head and neck malignancies; however, it invariably results in radiation-induced damage to normal cerebral tissue, culminating in radiation-induced brain injury (RBI). Despite extensive research on radiation-induced neuroinflammation, the association between RBI and the Th17 cell differentiation pathway remains inadequately understood. C57BL/6 mice underwent a single administration of 30 Gy cranial irradiation to develop the RBI model. Cognitive function was evaluated through the Morris water maze (MWM), open field test, novel object recognition test, and rotarod test. Histopathological alterations in brain tissue were analyzed using hematoxylin and eosin (H&E) staining. Immunofluorescence staining was employed to assess the activation of microglia (IBA-1) and astrocytes (GFAP). RNA sequencing was conducted to identify differentially expressed genes, while Simple Western and qPCR were utilized to examine key signaling molecules involved in Th17 cell differentiation. RBI mice demonstrated marked cognitive deficits, particularly in spatial learning and memory retention. Histological examination indicated activation of microglia and astrocytes within the cortex and hippocampus of irradiated mice. RNA sequencing analysis identified a significant enrichment of the Th17 cell differentiation pathway in the cortex of the RBI group. Further validation through Simple Western and qPCR analyses confirmed the upregulation of TGF-β, IL-6, RORγt, IL-17, and P-STAT3 in the cortex of RBI mice. These findings suggested that the Th17 cell differentiation pathway played a pivotal role in the pathogenesis of radiation-induced brain injury. Neuroinflammation mediated by Th17 cells may be a critical mechanism underlying radiation-induced cognitive dysfunction.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Precision Ultrasound-guided Stem Cell Delivery for Vascular Repair in Aortic Diseases. 精确超声引导干细胞输送用于主动脉疾病的血管修复。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-20 DOI: 10.3791/68433

Qiuyue Gao, Qing Xu, Xuejie Cao, Xian Xu, Qingbo Xu, Chen Zhang, Baoqi Yu

{"title":"Precision Ultrasound-guided Stem Cell Delivery for Vascular Repair in Aortic Diseases.","authors":"Qiuyue Gao, Qing Xu, Xuejie Cao, Xian Xu, Qingbo Xu, Chen Zhang, Baoqi Yu","doi":"10.3791/68433","DOIUrl":"https://doi.org/10.3791/68433","url":null,"abstract":"Aortic aneurysms and dissections rupture are life-threatening cardiovascular emergencies. Early prevention and control of disease progression are essential for improving patient outcomes. Vascular stem cells play a crucial role in the repair of vascular injuries across various vascular diseases. However, intravascular stem cell injection therapies encounter challenges in ensuring efficient engraftment of exogenous stem cells within the vascular wall. To explore a more effective and minimally invasive approach for delivering stem cells in aortic diseases, we developed an ultrasound-guided precision-targeted stem cell delivery technology. Specifically, the technique utilized an ultra-high-resolution small animal color Doppler ultrasound imaging system to precisely locate the vascular wall of the ascending aorta in aortic disease mouse models. Subsequently, 1 × 106 mCherry-labeled vascular stem cells mixed within 25 µL of Matrigel were injected using a 29 G needle. The progression of the disease was monitored by small animal ultrasound. Finally, the aorta was harvested and analyzed using 3D imaging with a light sheet microscope to track injected vascular stem cells. The above results demonstrate that the injected vascular stem cells were delivered accurately to the vascular wall of the ascending aorta with minimal invasiveness, thereby effectively promoting vascular repair in aortic aneurysms and dissections. This ultrasound-guided stem cell delivery technology offers a promising, minimally invasive method for stem cell transplantation, potentially controlling the progression of aortic aneurysms and dissections and other related cardiovascular diseases in clinics in the future.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585386","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Application of Intragastric Single-Incision Laparoscopy Surgery in the Management of Gastroesophageal Junction Gastrointestinal Stromal Tumors. 胃内单切口腹腔镜手术在胃食管交界区胃肠道间质瘤治疗中的应用。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-17 DOI: 10.3791/67663

Jia Bo Zheng, Ningyuan Chen, Yijia Lin, Junjie Liu, Lei Lian, Huashe Wang

{"title":"Application of Intragastric Single-Incision Laparoscopy Surgery in the Management of Gastroesophageal Junction Gastrointestinal Stromal Tumors.","authors":"Jia Bo Zheng, Ningyuan Chen, Yijia Lin, Junjie Liu, Lei Lian, Huashe Wang","doi":"10.3791/67663","DOIUrl":"https://doi.org/10.3791/67663","url":null,"abstract":"Gastrointestinal stromal tumors (GISTs) at the gastroesophageal junction (GEJ) pose significant surgical challenges due to their proximity to critical structures. Traditional laparoscopic techniques can lead to excessive tissue removal and complications such as gastroesophageal reflux or pyloric stenosis, while transgastric approaches increase the risks of tumor seeding and intraperitoneal infections. Intragastric single-incision laparoscopic surgery (I-SILS) offers a novel solution by combining intragastric visualization of the tumor and laparoscopic precision with single-port access, allowing for direct tumor visualization and resection while preserving GEJ function. This article presents a 43-year-old male patient with an endophytic GEJ GIST (46.8 mm × 33.9 mm × 30.0 mm in size), successfully treated using I-SILS, as a representative example. The procedure was completed in 42 min with minimal blood loss. The mucosal and submucosal defect was repaired using continuous sutures along the long axis of the stomach to maximize function preservation, and the patient experienced an uneventful recovery with no postoperative complications. The patient was reviewed one year after surgery, and no stenosis or reflux was observed. This approach provides several advantages, including improved tumor visualization, reduced risk of tumor seeding, and preservation of normal gastric and esophageal function. However, this technique is not suitable for all GISTs, particularly those larger than 5 cm. Patient selection and preoperative assessment are crucial. Further research, including large-scale, randomized controlled trials -- is needed to confirm the indications, safety, and effectiveness of I-SILS for GEJ GISTs.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Delineating the Metabolic Phenotype of Biopsy-Derived Kidney Cells. 描述活检肾细胞的代谢表型。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-17 DOI: 10.3791/65457

Masataka Kawamura, Catherine Parmentier, Lisa A Robinson, Markus Selzner, Caitriona M McEvoy, Julia M Murphy, Maya G Bozzo-Rey, Sarah Q Crome, Ana Konvalinka, Sergi Clotet-Freixas

{"title":"Delineating the Metabolic Phenotype of Biopsy-Derived Kidney Cells.","authors":"Masataka Kawamura, Catherine Parmentier, Lisa A Robinson, Markus Selzner, Caitriona M McEvoy, Julia M Murphy, Maya G Bozzo-Rey, Sarah Q Crome, Ana Konvalinka, Sergi Clotet-Freixas","doi":"10.3791/65457","DOIUrl":"https://doi.org/10.3791/65457","url":null,"abstract":"Kidney transplantation is the optimal treatment for end-stage kidney disease; however, transplanted kidneys are often lost prematurely, with up to 50% graft loss at 10 years post-transplant. One of the major causes of premature graft loss is the injury sustained by the graft at the time of transplantation, known as ischemia-reperfusion injury (IRI). Delayed graft function (DGF), defined as the need for dialysis in the first week post-transplant, is a manifestation of severe IRI that shows functional and histologic features of acute kidney injury (AKI). While the mechanisms driving AKI remain unclear, accumulating evidence suggests that altered metabolic function in the allograft mediates AKI and may be the reason for DGF. Thus, deciphering and monitoring the metabolic underpinnings of IRI will improve our capacity to diagnose and prevent AKI. This article describes a unique method to assess mitochondrial respiration (by means of oxygen consumption rate), glycolysis (extracellular acidification rate), and intracellular ATP levels in needle biopsy-derived kidney cell suspensions. The methodology has been optimized in healthy adult male pigs and validated in a porcine model of auto-transplantation. The approach presented has the potential to enhance the real-time assessment of kidney allograft viability in the clinic. Profiling metabolism in patient-derived biopsies may also uncover new biology in other metabolism-based kidney diseases.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585335","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Assessing Microglial Phagocytosis of Myelin Debris in vitro Under Repeated Magnetic Stimulation. 反复磁刺激下体外观察髓磷脂碎片小胶质细胞吞噬作用。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-17 DOI: 10.3791/67642

Chenyuan Zhai, Jili Cai, Mei Du, Yuchen Fei, Qi Wu

{"title":"Assessing Microglial Phagocytosis of Myelin Debris in vitro Under Repeated Magnetic Stimulation.","authors":"Chenyuan Zhai, Jili Cai, Mei Du, Yuchen Fei, Qi Wu","doi":"10.3791/67642","DOIUrl":"https://doi.org/10.3791/67642","url":null,"abstract":"Microglia, the resident phagocytes of the central nervous system (CNS), play a pivotal role in maintaining CNS integrity and homeostasis by removing damaged cells, cellular debris, and myelin remnants. The accumulation of myelin debris is implicated in a range of CNS disorders, including multiple sclerosis, Alzheimer's disease, traumatic brain injury, and spinal cord injury. The presence of myelin debris not only exacerbates neuroinflammation but also hampers the regenerative potential of myelin. Therefore, enhancing the ability of microglia to clear myelin debris through phagocytosis represents a promising therapeutic strategy. Magnetic stimulation has emerged as an innovative treatment modality for CNS diseases, with growing evidence suggesting its potential to promote microglial phagocytosis and support CNS recovery. To further elucidate the effects of magnetic stimulation on microglial clearance of myelin debris, we designed an in vitro experiment involving the co-culture of microglia and myelin debris. The co-culture was subjected to repetitive magnetic stimulation to assess its impact on microglial phagocytic activity in the context of CNS pathology.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Isolation and Culture of Primary Retinal Müller Cells from Sprague-Dawley (SD) Rats. SD （Sprague-Dawley）大鼠原代视网膜网膜<s:1>细胞的分离与培养。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-06-17 DOI: 10.3791/68129

Yunhua Tang, Yue Sun, Yongqi Mao, Wenyan Peng, Wenfeng Zhang, Fuwen Zhang

{"title":"Isolation and Culture of Primary Retinal Müller Cells from Sprague-Dawley (SD) Rats.","authors":"Yunhua Tang, Yue Sun, Yongqi Mao, Wenyan Peng, Wenfeng Zhang, Fuwen Zhang","doi":"10.3791/68129","DOIUrl":"https://doi.org/10.3791/68129","url":null,"abstract":"Retinal Müller cells (RMCs) play a crucial role in providing structural support and regulating various functions within the retina. As a core component of the retinal microenvironment, RMCs perform several vital functions. Through their abundant ion channels, ligands, receptors, transmembrane transporters, and enzyme systems, these cells contribute to neurotransmitter and trophic factor secretion, regulate retinal metabolism, and maintain water-ion homeostasis. Notably, RMCs have recently been identified as a significant source of endogenous retinal regenerative stem cells, offering novel therapeutic targets for the treatment of retinal degenerative diseases. Therefore, studying RMCs is essential for understanding the pathological mechanisms underlying retinal disorders. This study systematically establishes a standardized experimental protocol that includes trypsin digestion and purification of primary RMCs, morphological observation using inverted optical microscopy and hematoxylin and eosin (HE) staining, specific protein identification through immunofluorescence staining, and cell purity analysis via flow cytometry. This protocol serves as a valuable reference for both basic research and clinical applications related to RMCs, supporting the exploration of their mechanisms in retinal diseases and advancing the development of therapeutic strategies.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0