Methods in molecular biology最新文献_第4页

Human Skeletal Muscle Niche Formation and Analysis with Spatial RNA Sequencing. 人类骨骼肌生态位的形成和空间RNA测序分析。

Methods in molecular biology Pub Date : 2025-04-05 DOI: 10.1007/7651_2025_619

Ben Clock, Michael Hicks

引用次数: 0

Complex Spheroids as an Alternative: In Vivo-Like 3D Model for Investigating the Impact of Stromal Cells on the Radiation Response of Tumors. 复杂球体作为一种选择：研究基质细胞对肿瘤辐射反应影响的体内样3D模型。

Methods in molecular biology Pub Date : 2025-04-03 DOI: 10.1007/7651_2025_614

Hanna Sentek, Diana Klein

引用次数: 0

Evaluation of Adult Mouse Brain Neurogenic Niche Behavior Culturing Adult Mice Brain Slice In Vitro. 体外培养成年小鼠脑切片对成年小鼠脑神经源性生态位行为的评价。

Methods in molecular biology Pub Date : 2025-04-03 DOI: 10.1007/7651_2025_611

Maria Isabel Alonso, Sonia Martínez-Páramo, Francisco Lamus, Ángel Gato

{"title":"Evaluation of Adult Mouse Brain Neurogenic Niche Behavior Culturing Adult Mice Brain Slice In Vitro.","authors":"Maria Isabel Alonso, Sonia Martínez-Páramo, Francisco Lamus, Ángel Gato","doi":"10.1007/7651_2025_611","DOIUrl":"https://doi.org/10.1007/7651_2025_611","url":null,"abstract":"Adult brain neural precursors carry out their biological activity in specific areas in which they are able to self-renew and differentiate into neurons. This is due to a complex microenvironment of cellular interrelations in which soluble factors from the neighboring cells, vascular structures, and the content of the brain ventricle cavity (cerebrospinal fluid) play a key role. This cellular functional entity, known as the \"neurogenic niche,\" is able to generate new mature neurons, which are functionally integrated into the neuronal circuits of the adult mammal brain. The complexity of neurogenic niche signaling, which include biologically active molecules such as growth factors and morphogens, requires an experimental approach in order to create specific modifications of the biological activity of some of these molecules by means of a model of the active neurogenic niche, allowing an evaluation of neural precursor behavior.Here we describe the adaptation of an in vitro culture technique of adult brain slices with selected coronal sections, involving the two main brain neurogenic niches, the sub-ventricular zone (SVZ), and the hippocampus dentate gyrus, together with their associated sub-ependymal zone (SEZ). We explain certain examples of the experimental approach to modify neurogenic niche soluble signaling, implanting latex microbeads as a carrier for soluble signals. Additionally, we introduce an immune-cytochemical approach involving bromodeoxyuridine detection as a neural precursor cellular lineage tracer in combination with different molecular expressions, as a means of testing progressive states of neural precursor differentiation and neuronal maturation.This system represents a suitable strategy for evaluating the biological role of soluble components of the adult brain neurogenic niche.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143764542","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

Detection of Cellular Senescence on Murine Muscle Tissue Sections by Senescence-Associated β-Galactosidase Staining. 衰老相关β-半乳糖苷酶染色法检测小鼠肌肉组织切片细胞衰老。

Methods in molecular biology Pub Date : 2025-04-03 DOI: 10.1007/7651_2025_612

Wenxin Zhang, Tom H Cheung

引用次数: 0

Oligopaint FISH in Drosophila Testes. 果蝇睾丸中的低聚色素FISH。

Methods in molecular biology Pub Date : 2025-04-03 DOI: 10.1007/7651_2025_613

Romir Raj, Vedansh Patel, Mayu Inaba

引用次数: 0

Testicular Organoid Formation in Microwell Culture. 微孔培养中睾丸类器官的形成。

Methods in molecular biology Pub Date : 2025-03-28 DOI: 10.1007/7651_2025_624

Nathalia de Lima E Martins Lara, Anja Elsenhans, Ina Dobrinski

引用次数: 0

Organoid-Immune Cell Co-culture for Stable Live Imaging. 类器官-免疫细胞共培养稳定的活体成像。

Methods in molecular biology Pub Date : 2025-03-28 DOI: 10.1007/7651_2025_627

Nathalia Ferreira, Frauke Alves, Andrea Markus

引用次数: 0

Efficient Isolation and Ex Vivo Differentiation of Murine Satellite Cells from Healthy and Dystrophic Muscle. 从健康和萎缩性肌肉中高效分离和体外分化小鼠卫星细胞

Methods in molecular biology Pub Date : 2025-03-22 DOI: 10.1007/7651_2025_608

Alessio A Cusmano, Cordell A VanGenderen, Tim O Lorenz, Yafen Yang, Natasha C Chang

{"title":"Efficient Isolation and Ex Vivo Differentiation of Murine Satellite Cells from Healthy and Dystrophic Muscle.","authors":"Alessio A Cusmano, Cordell A VanGenderen, Tim O Lorenz, Yafen Yang, Natasha C Chang","doi":"10.1007/7651_2025_608","DOIUrl":"https://doi.org/10.1007/7651_2025_608","url":null,"abstract":"Satellite cells are the stem cells of adult skeletal muscles and confer skeletal muscle with remarkable regenerative ability. Under homeostatic conditions, satellite cells reside in a quiescent state in their niche along the basal lamina of muscle fibers. Upon receiving stimuli, satellite cells activate and engage in regenerative myogenesis to repair damaged fibers. Due to the impact of satellite cell differentiation on muscle physiology, studying their differentiation is relevant both within the context of healthy and diseased muscle. Due to the abundance of cell populations within skeletal muscle, the study of satellite cells is predicated on isolating highly pure populations. Fluorescence activated cell sorting (FACS) represents the gold standard for deriving highly pure satellite cell isolates but is costly and can reduce cell viability. In addition, proliferating satellite cells in vitro invariably transition to a homogeneous myoblast population that bestows a selective advantage on fast-dividing cells, reducing satellite cell heterogeneity. In this chapter, we describe our protocol for magnetic-activated cell sorting (MACS) of satellite cells. MACS preserves cell viability to a greater degree than FACS, and our approach allows for highly pure sorted populations of satellite cells. In addition, sorted cells can enter and progress through the myogenic program immediately upon plating, avoiding the need for lengthy expansion periods.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143674220","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

Matrix-Free Normal Human Epithelial-Fibroblast 3D Spheroid Cultures for In Vitro Lung Modeling. 无基质正常人上皮成纤维细胞三维球形体外肺模型培养。

Methods in molecular biology Pub Date : 2025-03-20 DOI: 10.1007/7651_2025_622

Lisa Marie Stasch, Maja Buchholzki, Zehra Sevindik, Bettina Budeus, Diana Klein

{"title":"Matrix-Free Normal Human Epithelial-Fibroblast 3D Spheroid Cultures for In Vitro Lung Modeling.","authors":"Lisa Marie Stasch, Maja Buchholzki, Zehra Sevindik, Bettina Budeus, Diana Klein","doi":"10.1007/7651_2025_622","DOIUrl":"https://doi.org/10.1007/7651_2025_622","url":null,"abstract":"The cellular responses of classical 2D flat monolayer cell culture systems provide only very limited reliable predictions about possible outcomes of corresponding animal experiments and clinical studies, which is due, among other things, to the lack of (bi)directional signaling transmission between different cell types and the lack of a structural microenvironment. To study the interactive communication between different cell types in vitro, two main co-culture methods have emerged as central techniques. In the indirect co-culture method, different cells are cultured physically separately (e.g., using transwell inserts) but can communicate with each other via secreted factors (paracrine mechanism). In the direct co-culture method, the different cells have direct physical contact, which enables direct interactions. Regarding the latter method, the cultivation of cells as spherical cell aggregates, so-called spheroids, embedded in a semi-solid extracellular matrix has been established as an in vivo-related, more complex cell culture model with different functional cell states according to cell-cell and cell-ECM interactions as well as oxygen and nutrient gradients. Here, we present a matrix-free method for direct spheroidal co-cultivation of human bronchial epithelial cells and fibroblasts, which can be considered as an in vivo-approximated cultivation method, especially with regard to the cellular composition of the respective spheroids.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143663697","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

Free-Floating Human Lung Organoids Derived from Induced Pluripotent Stem Cells. 诱导多能干细胞衍生的自由漂浮的人肺类器官。

Methods in molecular biology Pub Date : 2025-03-20 DOI: 10.1007/7651_2025_621

Bettina Budeus, Chiara Kroepel, Lisa Marie Stasch, Diana Klein

引用次数: 0