Methods in molecular biology最新文献_第4页

Direct Reprogramming of Human Fibroblasts into Fully Functional Trophoblast Stem Cells. 人类成纤维细胞直接重编程成功能齐全的滋养细胞干细胞。

Methods in molecular biology Pub Date : 2025-05-20 DOI: 10.1007/7651_2025_648

Meir Azagury, Yosef Buganim

引用次数: 0

Microfluidic On-Chip Assay for Quantifying Blastoid Adhesion to Endometrial Epithelium. 微流控芯片法定量测定囊胚与子宫内膜上皮的粘附。

Methods in molecular biology Pub Date : 2025-05-17 DOI: 10.1007/7651_2025_645

Aslı Ak, Dorian Luijkx, Daniel Carvalho, Stefan Giselbrecht, Ron van Golde, Erik Vrij

{"title":"Microfluidic On-Chip Assay for Quantifying Blastoid Adhesion to Endometrial Epithelium.","authors":"Aslı Ak, Dorian Luijkx, Daniel Carvalho, Stefan Giselbrecht, Ron van Golde, Erik Vrij","doi":"10.1007/7651_2025_645","DOIUrl":"https://doi.org/10.1007/7651_2025_645","url":null,"abstract":"The ability of the endometrium to accept and support embryo implantation is crucial, but factors influencing this process remain elusive. This method aims to obtain precise quantitative information on factors causally affecting the initial stages of embryo implantation. We developed a personalized implantation-on-chip platform using in vitro models of the endometrium (organoids) and the embryo (blastoids) to quantify functional embryo attachment. Here, we describe a microfluidic platform for precisely assessing functional receptivity of endometrial epithelium through blastoid adhesion. Endometrial organoids were expanded and transformed into epithelial monolayers within custom-made microfluidic chips. These chips were then infused with large numbers of blastoids (>100) per chip. Followed after 48 h of co-culture, blastoids were exposed to a controlled stepwise increasing flow rate (50, 100 and 400 μL/min), while the rate of adhered blastoids was precisely measured from image-based readouts. Our method offers a robust platform for studying endometrial epithelial receptivity and testing therapeutic interventions with potential impact for infertile patients.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144078791","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

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

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

Spheroid Invasion Assay of Melanoma Cells by Hanging Drop Technique. 挂滴法检测黑素瘤细胞球形浸润。

Methods in molecular biology Pub Date : 2025-03-20 DOI: 10.1007/7651_2025_615

Asiye Busra Boz Er, Ceren Sumer

引用次数: 0

Establishment of 3D Tooth Organoid Culture from Early-Postnatal Mouse Molar and Incisor. 生后早期小鼠磨牙和门牙三维类器官培养的建立。

Methods in molecular biology Pub Date : 2025-03-20 DOI: 10.1007/7651_2025_623

Florian Hermans, Hugo Vankelecom, Annelies Bronckaers, Ivo Lambrichts

引用次数: 0

Methodology for Assessing Drug Efficacy: Protocol for Single and Combination Drug Screening Using HeLa Cell Cultures. 评估药物疗效的方法学：使用HeLa细胞培养进行单一和联合药物筛选的方案。

Methods in molecular biology Pub Date : 2025-03-20 DOI: 10.1007/7651_2025_605

Phumuzile Dube, Bernice Monchusi, Mutsa M Takundwa, Vanelle L Kenmogne, Austin Malise, Deepak B Thimiri Govinda Raj

{"title":"Methodology for Assessing Drug Efficacy: Protocol for Single and Combination Drug Screening Using HeLa Cell Cultures.","authors":"Phumuzile Dube, Bernice Monchusi, Mutsa M Takundwa, Vanelle L Kenmogne, Austin Malise, Deepak B Thimiri Govinda Raj","doi":"10.1007/7651_2025_605","DOIUrl":"https://doi.org/10.1007/7651_2025_605","url":null,"abstract":"This protocol outlines a detailed method for performing drug sensitivity testing (DST) on HeLa cells, focusing on both single-drug and combination-drug screening to assess cell viability. DST is integral to cancer research and functional precision medicine, providing insight into individual drug responses and facilitating the optimization of drug combinations. The protocol includes preparing and maintaining HeLa cell cultures, seeding in 96-well plates, and performing single and combination drug treatments using a low-throughput screening approach. These drug treatments aim to evaluate therapeutic effectiveness, enhance understanding of synergistic interactions, and identify optimal combinations that could minimize toxicity while overcoming resistance. Data analysis uses open-source tools, including the BREEZE pipeline and Synergy-Finder, allowing for precise analysis of cell viability and drug interactions. This protocol provides a robust, reproducible framework for DST in cancer research, applicable to other cell lines, patient samples, and various drug types/classes. The critical role of DST in improving clinical treatment strategies through precise, scalable drug response analysis is demonstrated.","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":"143663736","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

Deep Learning-Driven Computational Approaches for Studying Intrinsically Disordered Regions in S100-A9. 基于深度学习驱动的S100-A9内部无序区域研究方法

Methods in molecular biology Pub Date : 2025-03-20 DOI: 10.1007/7651_2025_617

Gionathan L Distefano, Fabio D'Amico

引用次数: 0