Nature Protocols最新文献_第7页

Identification, assembly and characterization of tumor immunoglobulin transcripts from RNA sequencing data using IgSeqR. 利用IgSeqR从RNA测序数据中鉴定、组装和表征肿瘤免疫球蛋白转录物。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-24 DOI: 10.1038/s41596-025-01172-6

Dean Bryant, Benjamin Sale, Giorgia Chiodin, Dylan Tatterton, Benjamin Stevens, Alyssa Adlaon, Erin Snook, James Batchelor, Alberto Orfao, Francesco Forconi

{"title":"Identification, assembly and characterization of tumor immunoglobulin transcripts from RNA sequencing data using IgSeqR.","authors":"Dean Bryant, Benjamin Sale, Giorgia Chiodin, Dylan Tatterton, Benjamin Stevens, Alyssa Adlaon, Erin Snook, James Batchelor, Alberto Orfao, Francesco Forconi","doi":"10.1038/s41596-025-01172-6","DOIUrl":"https://doi.org/10.1038/s41596-025-01172-6","url":null,"abstract":"Immunoglobulin gene analysis provides fundamental insight into B cell receptor structure and function. In B cell tumors, it can provide information on the cell of origin and predict clinical outcomes. Its clinical value has been established in the two main types of chronic lymphocytic leukemia, which are distinguished by the expression of unmutated or mutated immunoglobulin heavy chain variable region (IGHV) genes, and is emerging in other B cell tumors. The traditional PCR and Sanger sequencing-based techniques for immunoglobulin gene analysis are labor-intensive and rely on attaining either a dominant sequence or a small number of subclonal sequences. Extraction of the expressed tumor immunoglobulin transcripts by using high-throughput RNA-sequencing (RNA-seq) can be faster, allow the collection of the tumor immunoglobulin sequence and match this with the rest of the RNA-seq data. Analytical tools are regularly sought to increase the accuracy, depth and speed of acquisition of the immunoglobulin transcript sequences and combine the immunoglobulin characteristics with other tumor features. We provide here a user-friendly protocol for the rapid (~1 h) de novo assembly, identification and accurate characterization of the full (leader to constant region) tumor immunoglobulin templated and non-templated transcript sequence from RNA-seq data ( https://github.com/ForconiLab/IgSeqR ). The derived amino acid sequences can be interrogated for their physicochemical characteristics and, in certain lymphomas, be used to predict tumor glycan types occupying acquired N-glycosylation sites. These features will then be available for association studies with the tumor transcriptome. The resulting information can also help refine diagnosis, prognosis and potential therapeutic targeting in the most common lymphomas.","PeriodicalId":18901,"journal":{"name":"Nature Protocols","volume":" ","pages":""},"PeriodicalIF":13.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144025240","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Empowering materials science with VASPKIT: a toolkit for enhanced simulation and analysis. 赋予材料科学与VASPKIT：一个工具包增强模拟和分析。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-23 DOI: 10.1038/s41596-025-01160-w

Wen-Tong Geng, Ya-Chao Liu, Nan Xu, Gang Tang, Yoshiyuki Kawazoe, Vei Wang

{"title":"Empowering materials science with VASPKIT: a toolkit for enhanced simulation and analysis.","authors":"Wen-Tong Geng, Ya-Chao Liu, Nan Xu, Gang Tang, Yoshiyuki Kawazoe, Vei Wang","doi":"10.1038/s41596-025-01160-w","DOIUrl":"https://doi.org/10.1038/s41596-025-01160-w","url":null,"abstract":"Driven by rapid advances in high-performance supercomputing, computational materials science has emerged as a powerful approach for exploring, designing, and predicting material properties at the atomic and molecular scales. Among the various computational tools developed in this field, the Vienna Ab initio Simulation Package (VASP) stands out as a widely adopted and highly versatile platform for performing first-principles density functional theory (DFT) calculations. VASP is widely used to explore electronic structures, phonon behavior, magnetic properties, thermodynamics and catalytic mechanisms across a diverse range of materials systems. Despite its robust capabilities, utilizing VASP requires expertise in setting up simulations and analyzing results, which can be time consuming and technically challenging. To address these barriers, VASPKIT was developed as a comprehensive toolkit to simplify the workflow for VASP users. VASPKIT streamlines both preprocessing and postprocessing tasks, enabling users to generate essential input files based on customizable parameters and automate computational workflows. The postprocessing features of VASPKIT allow for efficient analysis of electronic, mechanical, optical and catalytic properties, thereby substantially reducing the need for advanced programming expertise. This protocol provides a detailed guide to using VASPKIT, including practical examples to demonstrate its versatility and utility in conducting and analyzing DFT calculations. For instance, the computation of elastic constants, electronic band structures and density of states for a graphene system can typically be completed within half an hour, depending on the computational resources available. By offering step-by-step guidance, this protocol aims to further expand the accessibility and impact of VASPKIT in the field of computational materials science.","PeriodicalId":18901,"journal":{"name":"Nature Protocols","volume":" ","pages":""},"PeriodicalIF":13.1,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143972541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Direct analysis of biotransformations with mass spectrometry-DiBT-MS. 质谱- dibt - ms直接分析生物转化。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-21 DOI: 10.1038/s41596-025-01161-9

Ruth Knox, Rachel Smith, Emily E Kempa, Reynard Spiess, Christian Schnepel, Nicholas J Turner, Sabine L Flitsch, Perdita E Barran

{"title":"Direct analysis of biotransformations with mass spectrometry-DiBT-MS.","authors":"Ruth Knox, Rachel Smith, Emily E Kempa, Reynard Spiess, Christian Schnepel, Nicholas J Turner, Sabine L Flitsch, Perdita E Barran","doi":"10.1038/s41596-025-01161-9","DOIUrl":"https://doi.org/10.1038/s41596-025-01161-9","url":null,"abstract":"The development and analysis of engineered enzymes is greatly assisted by the use of high-throughput screening to quickly determine the efficacy of biotransformations under various conditions. Ambient ionization, particularly desorption electrospray ionization (DESI), coupled to high-resolution mass spectrometry has the advantages of minimal requirements for sample preparation before analysis, which renders it suitable for high-throughput screening, in which the accurate mass and potentially the tandem mass spectrometry (MS) fingerprint for any given product can be used for identification. We present a protocol that permits the application of this method in routine biotechnology and chemical biology laboratories that are using engineered enzymes (such as imine reductases and carboxylic acid reductases, mentioned herein) to produce target compounds from substrates (quinoline moieties and phenyl(piperazinyl) moieties, respectively). Through the use of DESI's MS imaging capabilities, reaction monitoring can be easily visualized via imaging of selected substrate or product ions in a convenient, user-friendly workflow. We describe here how DESI-MS can be used to directly analyze the activity of biotransformations from crude cell lysate, which we term 'DiBT-MS'. The DiBT-MS method presented here is 10-1,000 times as fast as liquid chromatography-MS, with the full procedure for 96 samples taking ~2 h and consuming far less solvent and sample. Also demonstrated in this protocol is the impact of solvent spray composition on ionization efficiency of the target analyte, the benefits of a nylon membrane slide and the reusability of sample slides in multiple experiments.","PeriodicalId":18901,"journal":{"name":"Nature Protocols","volume":" ","pages":""},"PeriodicalIF":13.1,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144025217","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Byproducts in the synthesis of [⁶⁸Ga]Ga-PSMA-11. [68Ga]Ga-PSMA-11合成副产物。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-18 DOI: 10.1038/s41596-025-01164-6

Ata Makarem, Jianlin Han, Karel D Klika

引用次数: 0

Reply to: Byproducts in the synthesis of [⁶⁸Ga]Ga-PSMA-11. [68Ga]Ga-PSMA-11合成副产物。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-18 DOI: 10.1038/s41596-025-01163-7

Andrew Katsifis, Daniela Stark, Michael J Fulham, Katherine Gagnon, Stefan Eberl, Peter J H Scott

引用次数: 0

A tripartite cell-free translation system to study mammalian translation. 研究哺乳动物翻译的三方无细胞翻译系统。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-16 DOI: 10.1038/s41596-025-01155-7

Frederic S W Arendrup, Kasper L Andersen, Anders H Lund

{"title":"A tripartite cell-free translation system to study mammalian translation.","authors":"Frederic S W Arendrup, Kasper L Andersen, Anders H Lund","doi":"10.1038/s41596-025-01155-7","DOIUrl":"https://doi.org/10.1038/s41596-025-01155-7","url":null,"abstract":"Genetic manipulation of cellular systems often leads to the adaptation of gene expression programs, rendering detailed mechanistic insights challenging to isolate and elucidate. The proteome constitutes the ultimate manifestation of gene expression programs with multiple layers of regulation to ensure faithful execution. While current high-throughput techniques to investigate regulation at the level of translation, such as Ribo-Seq and nascent proteomics, can capture nuanced changes in the translational landscape, they suffer from potential confounding factors imposed by adaptation of the cellular states. Cell-free translation systems have been used to elucidate molecular mechanisms for decades, but experimental setups have rigid composition and often rely on non-human model systems and artificially designed mRNA constructs. Here we detail a tripartite cell-free translation system based on the separation of mRNAs, ribosomes and ribosome-depleted cytoplasmic lysate from human cells, allowing for flexible reconstitution of translation reactions, which can be performed in 1-4 days. In this setup, cellular parts such as the cytoplasmic lysate can be kept constant, while ribosome complexes or mRNA can be varied or subjected to treatments or vice versa. We detail how complete mRNA populations can be used as input with subsequent detection of nascent peptides using autoradiography or mass spectrometry. We utilize this protocol to resolve which aspects of the translational machinery are selectively affected by environmental and cellular stress conditions that trigger ribosome stalling and collisions, which have been unresolvable until now.","PeriodicalId":18901,"journal":{"name":"Nature Protocols","volume":" ","pages":""},"PeriodicalIF":13.1,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143993947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

NIR-II aggregation-induced emission nanoparticles camouflaged with preactivated macrophage membranes for phototheranostics of pulmonary tuberculosis. 用预激活的巨噬细胞膜伪装NIR-II聚集诱导的发射纳米颗粒用于肺结核的光疗。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-10 DOI: 10.1038/s41596-025-01146-8

Dingyuan Yan, Xue Li, Huanhuan Wang, Bin Li, Wei Wang, Yuhui Liao, Ben Zhong Tang, Dong Wang

{"title":"NIR-II aggregation-induced emission nanoparticles camouflaged with preactivated macrophage membranes for phototheranostics of pulmonary tuberculosis.","authors":"Dingyuan Yan, Xue Li, Huanhuan Wang, Bin Li, Wei Wang, Yuhui Liao, Ben Zhong Tang, Dong Wang","doi":"10.1038/s41596-025-01146-8","DOIUrl":"https://doi.org/10.1038/s41596-025-01146-8","url":null,"abstract":"Phototheranostics, which allows simultaneous diagnosis and therapy, offers notable advantages in terms of noninvasiveness, controllability and negligible drug resistance, presenting a promising approach for disease treatment. By integrating second near-infrared (NIR-II, 1,000-1,700 nm) phototheranostic agents characterized by aggregation-induced emission (AIE) and cell membranes with specific targeting capacity, we have developed a versatile type of biomimetic nanoparticle (NP) for precise phototheranostics of pulmonary tuberculosis (TB). Coating the phototheranostic agents with preactivated macrophage membranes results in the formation of biomimetic NPs, which exhibit specific binding to TB through a lesion-pathogen dual-targeting strategy, allowing the accurate detection of Mycobacterium tuberculosis via NIR-II fluorescence imaging and precise photothermal therapy using the irradiation of a 1,064 nm laser. In comparison with traditional treatments, small individual granulomas (0.2 mm in diameter) in TB-infected mice are visualized, and improved antibacterial effects are achieved upon NP administration. Here we present a standardized workflow for the synthesis of the NIR-II AIE agents, their use for the fabrication of the biomimetic NPs and their in vitro and in vivo applications as phototheranostics against M. tuberculosis. The preparation and characterization of the NIR-II AIE agents requires ~8 d, the synthesis and characterization of the phototheranostic NPs requires ~8 d, the validation of in vitro targeting capacity and photothermal eradication requires ~26 d, and the in vivo NIR-II fluorescence imaging and imaging-guided photothermal therapy requires ~74 d. All procedures are straightforward and suitable for clinicians or researchers with prior training in organic synthesis and biomedical engineering.","PeriodicalId":18901,"journal":{"name":"Nature Protocols","volume":" ","pages":""},"PeriodicalIF":13.1,"publicationDate":"2025-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144019785","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Transcriptomic profiling of individual bacteria by MATQ-seq. 利用MATQ-seq分析单个细菌的转录组学特征。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-09 DOI: 10.1038/s41596-025-01157-5

Christina Homberger, Fabian Imdahl, Regan J Hayward, Lars Barquist, Antoine-Emmanuel Saliba, Jörg Vogel

{"title":"Transcriptomic profiling of individual bacteria by MATQ-seq.","authors":"Christina Homberger, Fabian Imdahl, Regan J Hayward, Lars Barquist, Antoine-Emmanuel Saliba, Jörg Vogel","doi":"10.1038/s41596-025-01157-5","DOIUrl":"10.1038/s41596-025-01157-5","url":null,"abstract":"Bacterial single-cell transcriptomics is revolutionizing our understanding of cell-to-cell variation within bacterial populations and enables gene expression profiling in complex microbial communities. Using the eukaryotic multiple annealing and dC-tailing-based quantitative single-cell RNA-sequencing (scRNA-seq) (MATQ-seq) approach, we have developed a robust bacterial scRNA-seq protocol, which integrates index sorting, random priming and rRNA depletion. This method stands out for its high rate of cell retention and its suitability for experiments with limited input material, offering a reliable method even for small sample sizes. Here we provide a step-by-step protocol covering the entire process of generating single-bacteria transcriptomes, including experimental and computational analysis. It involves (i) single-cell isolation via fluorescence-activated cell sorting (FACS) and cell lysis, (ii) reverse transcription and cDNA amplification using robotic liquid handling, (iii) rRNA depletion, (iv) indexing and sequencing, and (v) data processing steps to start comprehensive data analysis. Using model organisms such as Salmonella enterica, we show that the method achieves a retention rate of 95%, defined as the rate of initially sorted cells converted into effective sequencing libraries. This substantially surpasses other available protocols. The method robustly detects 300-600 genes per cell, highlighting its effectiveness in capturing a broad transcriptomic profile. The entire procedure from FACS-based single-cell isolation to raw data generation spans ~5 d. As MATQ-seq has already been proven robust in several bacterial species, it holds promise for the establishment of a streamlined microbial scRNA-seq platform.","PeriodicalId":18901,"journal":{"name":"Nature Protocols","volume":" ","pages":""},"PeriodicalIF":13.1,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144044912","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Sequential orthogonal assays for longitudinal and endpoint characterization of three-dimensional spheroids. 三维球体纵向和终点特征的顺序正交试验。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-08 DOI: 10.1038/s41596-025-01150-y

Eva Blondeel, Sam Ernst, Felix De Vuyst, Ákos Diósdi, Cláudio Pinheiro, Diogo Estêvão, Pekka Rappu, Robin Boiy, Sándor Dedeyne, Ligia Craciun, Vera Goossens, Jonas Dehairs, Tânia Cruz, Dominique Audenaert, Wim Ceelen, Michael Linnebacher, Tom Boterberg, Jo Vandesompele, Pieter Mestdagh, Johan Swinnen, Jyrki Heino, Peter Horvath, Maria José Oliveira, An Hendrix, Pieter Demetter, Olivier De Wever

{"title":"Sequential orthogonal assays for longitudinal and endpoint characterization of three-dimensional spheroids.","authors":"Eva Blondeel, Sam Ernst, Felix De Vuyst, Ákos Diósdi, Cláudio Pinheiro, Diogo Estêvão, Pekka Rappu, Robin Boiy, Sándor Dedeyne, Ligia Craciun, Vera Goossens, Jonas Dehairs, Tânia Cruz, Dominique Audenaert, Wim Ceelen, Michael Linnebacher, Tom Boterberg, Jo Vandesompele, Pieter Mestdagh, Johan Swinnen, Jyrki Heino, Peter Horvath, Maria José Oliveira, An Hendrix, Pieter Demetter, Olivier De Wever","doi":"10.1038/s41596-025-01150-y","DOIUrl":"10.1038/s41596-025-01150-y","url":null,"abstract":"Spheroids are reaggregated multicellular three-dimensional structures generated from cells or cell cultures of healthy as well as pathological tissue. Basic and translational spheroid application across academia and industry have led to the development of multiple setups and analysis methods, which mostly lack the modularity to maximally phenotype spheroids. Here we present the self-assembly of single-cell suspensions into spheroids by the liquid overlay method, followed by a modular framework for a multifaceted phenotyping of spheroids. Cell seeding, supernatant handling and compound administration are elaborated by both manual and automated procedures. The phenotyping modules contain a suite of orthogonal assays to analyze spheroids longitudinally and/or at an endpoint. Longitudinal analyses include morphometry with or without spheroid or cell state specific information and supernatant evaluation (nutrient consumption and metabolite/cytokine production). Spheroids can also be used as a starting point to monitor single and collective cell migration and invasion. At an endpoint, spheroids are lysed, fixed or dissociated into single cells. Endpoint analyses allow the investigation of molecular content, single-cell composition and state and architecture with spatial cell and subcellular specific information. Each module addresses time requirements and quality control indicators to support reproducibility. The presented complementary techniques can be readily adopted by researchers experienced in cell culture and basic molecular biology. We anticipate that this modular protocol will advance the application of three-dimensional biology by providing scalable and complementary methods.","PeriodicalId":18901,"journal":{"name":"Nature Protocols","volume":" ","pages":""},"PeriodicalIF":13.1,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143811809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Human induced pluripotent stem cell-derived cardiomyocytes and their use in a cardiac organ-on-a-chip to assay electrophysiology, calcium and contractility. 人诱导多能干细胞衍生的心肌细胞及其在心脏器官芯片上测定电生理、钙和收缩性的应用。

IF 13.1 1区生物学

Nature Protocols Pub Date : 2025-04-07 DOI: 10.1038/s41596-025-01166-4

M Iveth Garcia, Keri Dame, Verena Charwat, Brian A Siemons, Henrik Finsberg, Bhavya Bhardwaj, Ryosuke Yokosawa, Ishan Goswami, Dylan Bruckner, Samuel T Wall, Kevin A Ford, Kevin E Healy, Alexandre J S Ribeiro

{"title":"Human induced pluripotent stem cell-derived cardiomyocytes and their use in a cardiac organ-on-a-chip to assay electrophysiology, calcium and contractility.","authors":"M Iveth Garcia, Keri Dame, Verena Charwat, Brian A Siemons, Henrik Finsberg, Bhavya Bhardwaj, Ryosuke Yokosawa, Ishan Goswami, Dylan Bruckner, Samuel T Wall, Kevin A Ford, Kevin E Healy, Alexandre J S Ribeiro","doi":"10.1038/s41596-025-01166-4","DOIUrl":"https://doi.org/10.1038/s41596-025-01166-4","url":null,"abstract":"Cardiac organs-on-a-chip (OoCs) or microphysiological systems have the potential to predict cardiac effects of new drug candidates, including unanticipated cardiac outcomes, which are among the main causes for drug attrition. This protocol describes how to prepare and use a cardiac OoC containing cardiomyocytes differentiated from human induced pluripotent stem cells (hiPS cells). The use of cells derived from hiPS cells as reliable sources of human cells from diverse genetic backgrounds also holds great potential, especially when cultured in OoCs that are physiologically relevant culture platforms. To promote the broad adoption of hiPS cell-derived cardiac OoCs in the drug development field, there is a need to first ensure reproducibility in their preparation and use. This protocol aims to provide key information on how to reduce sources of variability during hiPS cell maintenance, differentiation, loading and maturation in OoCs. Variability in these procedures can lead to inconsistent purity after differentiation and variable function between batches of microtissues formed in OoCs. This protocol also focuses on describing the handling and functional assessment of cardiac microtissues using live-cell microscopy approaches to quantify parameters of cellular electrophysiology, calcium transients and contractility. The protocol consists of five stages: (1) thaw and maintain hiPS cells, (2) differentiate hiPS cell cardiomyocytes, (3) load differentiated cells into OoCs, (4) maintain and characterize loaded cells, and (5) evaluate and utilize cardiac OoCs. Execution of the entire protocol takes ~40 days. The required skills to carry out the protocol are experience with sterile techniques, mammalian cell culture and maintaining hiPS cells in a pluripotent state.","PeriodicalId":18901,"journal":{"name":"Nature Protocols","volume":" ","pages":""},"PeriodicalIF":13.1,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0