Methods in molecular biology最新文献_第3页

From Single Stem Cells to an In Vitro Model of the Post-implantation Human Embryo: A Step-by-Step Guide. 从单个干细胞到植入后人类胚胎的体外模型：一步一步的指南。

Methods in molecular biology Pub Date : 2025-05-22 DOI: 10.1007/7651_2025_647

Seher Ipek Gassaloglu, Monique Pedroza, Berna Sozen

引用次数: 0

Linking Single-Cell Dynamics to Cell Fate in Differentiating hPSCs. 分化造血干细胞中单细胞动力学与细胞命运的联系。

Methods in molecular biology Pub Date : 2025-05-22 DOI: 10.1007/7651_2025_638

Seth Teague, Zhiyuan Yu, Idse Heemskerk

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Preparation and Characterization of Nanoparticles for the Controlled Release of the Endocannabinoid 2-Arachidonoylglycerol (2-AG). 内源性大麻素2-花生四烯醇甘油（2-AG）控释纳米颗粒的制备与表征

Methods in molecular biology Pub Date : 2025-05-22 DOI: 10.1007/7651_2025_636

Sevil Köse, Cem Varan, Tuba Reçber, Emirhan Nemutlu, Erem Bilensoy, Petek Korkusuz

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Isolation of Mouse Bone Marrow Niche Cells for Single-Cell Sequencing. 小鼠骨髓小生境细胞的分离及单细胞测序。

Methods in molecular biology Pub Date : 2025-05-22 DOI: 10.1007/7651_2025_630

Yaphet Bustos, Adrienne M Dorrance, Chinmayee Goda

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Engineering Synthetic PEG Hydrogels to Model Peri-implantation Epiblast Morphogenesis with Tunable Biophysical Properties. 工程合成聚乙二醇水凝胶以可调生物物理性质模拟植入周外胚层形态发生。

Methods in molecular biology Pub Date : 2025-05-22 DOI: 10.1007/7651_2025_640

Michael Patrick Seitz, Zhen Ma, Era Jain

{"title":"Engineering Synthetic PEG Hydrogels to Model Peri-implantation Epiblast Morphogenesis with Tunable Biophysical Properties.","authors":"Michael Patrick Seitz, Zhen Ma, Era Jain","doi":"10.1007/7651_2025_640","DOIUrl":"https://doi.org/10.1007/7651_2025_640","url":null,"abstract":"Implantation triggers critical morphological transformations in the embryo, where the epiblast transitions from a cluster of unpolarized cells into a highly organized, polarized epithelium characterized by a central lumen. Human pluripotent stem cells (hPSCs) are valuable models for studying this process, but conventional matrices like Matrigel have significant limitations, including variability and poor control over mechanical properties. To overcome these challenges, we developed a synthetic polyethylene glycol (PEG) hydrogel system with tunable mechanical stiffness to model peri-implantation epiblast morphogenesis.Our platform enables hPSCs to form unpolarized 3D aggregates that undergo stiffness-dependent transformation into lumen-forming, apicobasal-polarized structures resembling epiblast morphogenesis during peri-implantation. Unlike natural ECMs, PEG hydrogels maintain hPSC pluripotency for extended periods and support trilineage differentiation upon induction. The modular hydrogel design facilitates targeted mechanistic studies on the biophysical and biochemical regulation of cell morphogenesis. We present a comprehensive protocol for fabricating PEG hydrogels, encapsulating hPSCs, and assessing cell polarity, lumen formation, and pluripotency using immunostaining and RT-PCR. This platform provides a robust, cost-effective, and versatile tool for advancing developmental biology and regenerative medicine.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144111214","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

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

A Robust Human Liver Organoid Model of Hepatitis B Virus Infection. 乙型肝炎病毒感染的人类肝类器官模型。

Methods in molecular biology Pub Date : 2025-04-15 DOI: 10.1007/7651_2025_626

Bang M Tran, Linda Earnest, Dustin J Flanagan, Jean M Moselen, Hoanh Tran, Joseph Torresi, Elizabeth Vincan

引用次数: 0

Three-Dimensional Analysis of Skeletal Muscle Stem Cell Niche Following Tissue Clearing. 组织清除后骨骼肌干细胞生态位的三维分析

Methods in molecular biology Pub Date : 2025-04-05 DOI: 10.1007/7651_2025_618

Yoko Asakura, Smrithi Karthikeyan, Atsushi Asakura

{"title":"Three-Dimensional Analysis of Skeletal Muscle Stem Cell Niche Following Tissue Clearing.","authors":"Yoko Asakura, Smrithi Karthikeyan, Atsushi Asakura","doi":"10.1007/7651_2025_618","DOIUrl":"10.1007/7651_2025_618","url":null,"abstract":"Skeletal muscle is an intricately structured tissue made up of a complex framework of various cell types. The dynamic spatial and temporal relationships among these cells during both homeostasis and periods of injury contribute to the regenerative abilities of skeletal muscle. Currently, there is a deficiency in quantitative assessment, biological role, and the molecular mechanisms that could elucidate a possible juxtavascular niche for muscle satellite cells, a stem cell population for skeletal muscle regeneration. To fully comprehend the regeneration process by muscle satellite cells, a three-dimensional (3-D) imaging approach is essential. Confocal microscopy serves as an exceptional method for examining the spatial arrangement of cells within a specific tissue. In this protocol, we provide a detailed procedure for preparing optically transparent extensor digitorum longus (EDL) skeletal muscle specimens that are appropriate for confocal microscopy and computational 3-D assessment. We outline the steps for sample preparation, which include perfusion fixation and the tissue clearing process for rodent muscle specimens, as well as guidelines for image capture and computational evaluation featuring sample segmentation and 3-D visualization. This methodology can be utilized to characterize diverse cell types, such as muscle satellite cells and capillary endothelial cells found in rodent skeletal muscle.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143780471","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

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