Current protocols最新文献

{"title":"Surface Plasmon Resonance (SPR) Workflow for Comparative Analysis of Nanobody Variants Binding to Lysozyme as a Model Ligand.","authors":"Escarlet Díaz-Galicia, Nicoleta Gutu, Yuli Peng, Almira Valitova, Dominik Renn, Magnus Rueping","doi":"10.1002/cpz1.70360","DOIUrl":"10.1002/cpz1.70360","url":null,"abstract":"<p><p>Developing protein interaction-based technologies such as biosensors requires a clear understanding of receptor-target kinetics. Nanobodies, which are camelid-derived single-domain antibodies, are ideal biosensor receptors due to their high specificity, stability, and ease of production. During biosensor development, multiple nanobody variants are often tested against the same target to identify the best binders. Surface plasmon resonance (SPR) is a robust, label-free method for measuring these interactions, but its many experimental variables can complicate the establishment of a streamlined protocol specially for non-high-throughput instruments. Here, we present an SPR workflow that enables the comparative analysis of nanobody variants binding to lysozyme as a model target on a Biacore T100 instrument. These protocols cover the steps from protein expression and purification to final affinity ranking. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Expression and purification of lysozyme-specific nanobodies Support Protocol 1: Protein identification Basic Protocol 2: SPR workflow for the characterization of nanobody variants Support Protocol 2: SPR planning and assay development.</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 5","pages":"e70360"},"PeriodicalIF":2.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13127243/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147792789","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immortalization of Human Adult Alveolar Epithelial Cells to Study Environmental Exposures of the Distal Lung.","authors":"Evelyn Tran, Laurence St Pierre, Diego Velarde, Bianca Dal Bó, Juan Ramon Alvarez, Kweku Ofosu-Asante, Nazarius S Lamango, Yong Huang, Crystal N Marconett, Zea Borok, Beiyun Zhou, Ite A Offringa","doi":"10.1002/cpz1.70372","DOIUrl":"10.1002/cpz1.70372","url":null,"abstract":"<p><p>Alveolar epithelial cell type 1 (AT1) and type 2 (AT2) cells make up the saccular gas exchange units of the lung, called alveoli. Formation of alveoli during lung development accounts for the expansive surface area of the lung, allowing for proper respiration and delivery of oxygen to the body. Due to their delicate structure, alveoli are susceptible to injury caused by environmental exposures, such as inhaled cigarette smoke and heavy metals. Chronic exposure to these toxicants can exacerbate preexisting conditions such as asthma or contribute to the progression of lung diseases, such as chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), and cancer. AT2 cells play a key role in injury repair and regeneration of the distal lung and there is widespread interest in their use as cellular models for lung disease in vitro. However, while immortalized cell lines derived from airway epithelial cells were successfully generated decades ago, human adult alveolar epithelial cell lines have proven to be more difficult to establish due to their limited proliferative capacity. Here we describe an extensive end-to-end method for deriving immortalized alveolar epithelial cells (AECs), termed \"AEC-tLgT cells,\" from normal human lung tissue. We first outline a detailed procedure to isolate AT2 cells. We then outline our optimized method for immortalizing AT2 cells to generate polyclonal cell lines. We next describe a three-dimensional co-culture system to induce lung organoid formation from immortalized AT2 cell lines. Finally, we describe a procedure for studying cigarette smoke and nickel exposure using immortalized AT2 cell lines to investigate environmental toxicity. These protocols will enable users to generate AEC models from donors with defined genetic, demographic, and clinical backgrounds, facilitating the study of differential susceptibility to environmental exposures and risk for distal lung diseases. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Isolation and purification of human AT2 cells Basic Protocol 2: Immortalization of AT2 cells and maintenance of resulting AEC lines Basic Protocol 3: Determination of organoid formation ability Basic Protocol 4: Treatment of immortalized AEC lines to study environmental exposures.</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 5","pages":"e70372"},"PeriodicalIF":2.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13130368/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147792748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient Adaptation of Structure-Aware Protein Language Models for Diverse Protein Applications.","authors":"Duolin Wang, Yichuan Zhang, Yongfang Qin, Qing Shao, Dong Xu","doi":"10.1002/cpz1.70377","DOIUrl":"https://doi.org/10.1002/cpz1.70377","url":null,"abstract":"<p><p>Protein language models (PLMs) trained solely on sequence data have significantly advanced our understanding of protein biology and achieved remarkable performance in protein prediction tasks. However, their lack of three-dimensional (3D) structural features limits their predictive power in applications that rely heavily on 3D conformation. To address this limitation, we developed two structure-aware PLMs, S-PLM1 and S-PLM2, that employ multi-view contrastive learning to align protein sequences with their 3D structures in a unified latent space. S-PLM1 represents structural information using contact maps encoded by a pretrained Swin-Transformer, while S-PLM2 directly encodes 3D backbone coordinates through a Geometric Vector Perceptron (GVP)-based model. The paired sequence-structure data were obtained from AlphaFoldDB. For both models, we designed efficient tuning strategies that enable optimal performance with minimal computational cost. Here, we present detailed protocols for adapting S-PLM1 and S-PLM2 for diverse protein applications. The protocols provide step-by-step guidance on generating structure-aware representations from S-PLMs, fine-tuning them for various protein prediction tasks, and using S-PLM2 to produce structure embeddings for structure-based downstream analyses. We also provide source code and Google Colab implementations for easy customization and deployment. © 2026 Wiley Periodicals LLC. Basic Protocol 1: Generating structure-aware representations of protein sequences Basic Protocol 2: Efficient tuning of structure-aware protein language models for diverse protein applications Basic Protocol 3: Using S-PLM2 to generate protein structure representations and conduct structure-based clustering Support Protocol: Google Colab quick start notebooks.</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 5","pages":"e70377"},"PeriodicalIF":2.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147864813","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}

{"title":"Affordable CRISPR RNP-Based Genome Editing in Euglena gracilis.","authors":"Anzu Minami, Minami Shimizu, Shun Tamaki, Vicki Nishinarizki, Yosua, Keiichi Mochida","doi":"10.1002/cpz1.70357","DOIUrl":"https://doi.org/10.1002/cpz1.70357","url":null,"abstract":"<p><p>Genome editing can enhance basic research and enable industrial applications of green algae. Here, we present an affordable, broadly applicable workflow for genome editing in the unicellular green alga Euglena gracilis using Cas9 nucleases. This method retains high editing efficiency while significantly lowering technical barriers. Unlike previous approaches that required specialized equipment, this protocol can be performed using a general-purpose laboratory electroporator and a simplified clonal isolation procedure without the need for specialized micromanipulation devices. This protocol is compatible with a range of editing outcomes, such as targeted deletions and precise base substitutions, enabling more widespread genome editing in Euglena. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Culture of Euglena gracilis Basic Protocol 2: sgRNA synthesis Basic protocol 3: Transformation Basic protocol 4: Genotyping.</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 5","pages":"e70357"},"PeriodicalIF":2.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13142649/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147847779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Absolute Quantification of microRNA Copies in Exosomes Using Real-Time PCR","authors":"Pragati Raghuwanshi,&nbsp;Prasenjit Srivastava,&nbsp;Nishant Singh,&nbsp;Prabhat Kumar,&nbsp;Ashutosh Kumar Mishra,&nbsp;Archana Verma,&nbsp;Saroj Kumar,&nbsp;Avindra Nath,&nbsp;Sanjay Yadav","doi":"10.1002/cpz1.70352","DOIUrl":"https://doi.org/10.1002/cpz1.70352","url":null,"abstract":"<p>Exosomes are small extracellular vesicles (50 to 150 nm in size) that carry different kind of biomolecules in their lumen and facilitate intercellular communication. Serum contains a large number of exosomes (∼10¹²/ml of serum) that contain relatively more small RNAs than large RNAs. Quantifying the absolute number of specific miRNA molecules in exosomes is critical for understanding their functional relevance and developing them for diagnostic and therapeutic applications. This article describes an exosome copy number assay that combines nanoparticle tracking analysis (NTA) to quantify vesicle concentration with absolute quantification of miRNA using a standard curve-based qPCR approach. The workflow includes optimized procedures for vesicle isolation, particle measurement, RNA extraction, and absolute quantification, enabling calculation of miRNA copy number per exosome. The method can be used to generate reproducible and accurate results across diverse biological samples. By offering precise molecular quantification, these protocols provide a practical framework to advance exosome biology and support the development of exosome-based biomarkers and therapies. © 2026 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Isolation of exosomes from human serum by ultracentrifugation</p><p><b>Support Protocol</b>: Exogenous spike-in controls and troubleshooting</p><p><b>Basic Protocol 2</b>: NTA for exosome enumeration</p><p><b>Basic Protocol 3</b>: Extraction of RNA from purified exosomes from serum</p><p><b>Basic Protocol 4</b>: Reverse transcription (cDNA synthesis and preamplification) and real-time PCR setup</p><p><b>Basic Protocol 5</b>: Copy number assay (standard curve preparation)</p><p><b>Basic Protocol 6</b>: Calculation of miRNA copy number per vesicle (absolute quantification by qPCR and NTA normalization)</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147668529","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}

{"title":"Cellular Characterization Using Microwave-Assisted Immunocytochemistry, A Time-Efficient Alternative to Traditional Immunocytochemistry","authors":"Aidyn M. Medina-López,&nbsp;Irma I. Torres-Vázquez,&nbsp;Noraida Martínez-Rivera,&nbsp;Eduardo Rosa-Molinar","doi":"10.1002/cpz1.70363","DOIUrl":"https://doi.org/10.1002/cpz1.70363","url":null,"abstract":"<p>Immunocytochemistry (ICC) and immunohistochemistry (IHC) are related but separate techniques used for cell and tissue characterization. ICC involves immunolabeling of cells, and IHC immunolabeling of tissues; thus, the main difference is sample thickness. However, the distinction between ICC and IHC is not always clear and can be confusing. We show that a clear distinction should be made between the two. Importantly, we show that assisted ICC is an excellent method for characterizing cell lines. It is quick, reduces background noise without disrupting the expression and organization of the protein of interest, and helps ensure the rigor and reproducibility of results. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Culture of blood-spinal cord barrier and neurovascular unit cellular components</p><p><b>Basic Protocol 2</b>: Immunocytochemistry assisted by PELCO BioWave® Pro (Fig. 3)</p><p><b>Alternate Protocol</b>: Conventional immunocytochemistry</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://currentprotocols.onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70363","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147668526","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acid-Catalyzed Cycloaddition of Guanines with TMOP: A Unified Strategy for Synthesizing Tricyclic Acyclovir Analogues and M1G-Containing Oligonucleotides","authors":"Zhuo-Yue Jia,&nbsp;Bin Wu,&nbsp;Wen-Wu Sun","doi":"10.1002/cpz1.70362","DOIUrl":"10.1002/cpz1.70362","url":null,"abstract":"<p>This article describes an efficient acid-catalyzed cyclization strategy for constructing the tricyclic pyrimido[1,2-a]purin-10(3H)-one scaffold via the reaction of guanine derivatives with 1,1,3,3-tetramethoxypropane (TMOP) catalyzed by 2,4,5-trifluorobenzoic acid. The method features mild conditions, eliminates the need for hydroxyl protection, and exhibits excellent functional-group tolerance, making it applicable to the late-stage structural diversification of various nucleosides, nucleotides, and oligonucleotides. Using the antiviral drug acyclovir and an azide-containing guanosine derivative as model substrates, we successfully synthesized a tricyclic acyclovir analogue in 81% yield and an M₁G-containing oligonucleotide in 40% yield. This protocol provides a general, convenient, and efficient experimental approach for synthesizing this class of potentially bioactive tricyclic nucleoside analogues. © 2026 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Synthesis of tricyclic acyclovir analogue</p><p><b>Basic Protocol 2</b>: Synthesis of M₁G-containing oligonucleotide</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147635443","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}

{"title":"Co-Culture Systems to Study Epithelial-Immune Interactions During SARS-CoV-2 Infection","authors":"Scott H. Randell,&nbsp;Katherine C. Barnett","doi":"10.1002/cpz1.70365","DOIUrl":"10.1002/cpz1.70365","url":null,"abstract":"<p>Robust inflammatory responses to viral infection are mediated by immune cell populations, including monocytes and dendritic cells. However, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) does not replicate efficiently in these cell types and instead preferentially infects epithelial cell subsets in the airway. Because of this, stimulation of inflammatory cytokine responses from immune cell populations during SARS-CoV-2 infection depends not only on interactions with viral particles but also interactions with infected epithelial cells. In this article, we describe two co-culture systems to study inflammatory cytokine responses generated by epithelial-immune interaction during SARS-CoV-2 infection in vitro. Basic Protocol 1 describes how to set up a partially primary co-culture system consisting of SARS-CoV-2-infected Vero-E6 cells and primary human peripheral blood mononuclear cells (PBMCs) to observe release of the inflammasome-regulated cytokine interleukin-1β (IL-1β). Basic Protocol 2 details a primary, human co-culture system that consists of SARS-CoV-2-infected primary human airway epithelia (HAE) grown at an air–liquid interface (ALI) and primary human PBMCs, and how to observe IL-1β and IL-6 crosstalk between these cell populations during infection. In these Basic Protocols, we include a description of the use of inhibitors in these systems to perturb cytokine responses. We also provide Support Protocols for the culture of HAE and Vero-E6 and for the isolation, storage, and preparation of PBMCs prior to use in these systems. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: SARS-CoV-2 infection in a Vero-E6+PBMC co-culture system</p><p><b>Support Protocol 1</b>: Vero-E6 culture and passaging</p><p><b>Support Protocol 2</b>: Isolation and cryopreservation of PBMCs for use in co-culture</p><p><b>Basic Protocol 2</b>: SARS-CoV-2 infection in a primary HAE+PBMC co-culture system</p><p><b>Support Protocol 3</b>: Establishment and maturation of HAE grown at an ALI</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13060367/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147635449","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis of 2′,4′- and 3′,4′-Bridged Nucleosides via 4′-Carbon Radicals Generated by H-Abstraction","authors":"Yuta Ito,&nbsp;Yoshiyuki Hari","doi":"10.1002/cpz1.70353","DOIUrl":"10.1002/cpz1.70353","url":null,"abstract":"<p>This unit describes a method for the synthesis of 2′,4′- and 3′,4′-bridged nucleosides from nucleosides bearing oxime imidate moieties at the 5′ position. Hydrogen abstraction at the 4′-position of nucleosides by iminyl radicals derived from oxime imidates facilitates the facile generation of 4′-carbon radicals, which undergo cyclization and subsequent hydrolysis of the imidate residues to afford bridged nucleosides. Using this method, the eight-step syntheses of <i>S</i>-constrained ethyl-bridged 5-methyluridine (<i>S</i>-cEt-T) and 6′<i>S</i>-methyl-2′-<i>O</i>,4′-<i>C</i>-ethylene-bridged 5-methyluridine (6′<i>S</i>-Me-ENA-T), which are 2′,4′-bridged nucleosides with five- and six-membered bridges, respectively, were achieved. Moreover, 3′-<i>O</i>,4′-<i>C</i>-bridged 2′-deoxynucleosides bearing various bases were prepared in five steps. Overall, this H-abstraction/cyclization strategy provides an efficient route for constructing 2′,4′- and 3′,4′-bridges in fewer steps than with existing methods. © 2026 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Synthesis of <i>S</i>-constrained ethyl-bridged 5-methyluridine (<i>S</i>-cEt-T)</p><p><b>Basic Protocol 2</b>: Synthesis of 6′<i>S</i>-methyl-2′-<i>O</i>,4′-<i>C</i>-ethylene-bridged 5-methyluridine (6′<i>S</i>-Me-ENA-T)</p><p><b>Basic Protocol 3</b>: Synthesis of 3′-<i>O</i>,4′-<i>C</i>-bridged nucleosides</p><p><b>Support Protocol</b>: Preparation of (<i>Z</i>)-<i>N</i>-phenoxy-4-(trifluoromethyl)benzimidoyl chloride</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147629526","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}

{"title":"A Tutorial for Isolating, Characterizing, and Inducing Presenescence in Human Periodontal Ligament and Dental Pulp Stem Cells","authors":"Kamila Sauer Veiga Leme,&nbsp;Márjorie de Assis Golim,&nbsp;Aline Márcia Marques Braz,&nbsp;Elenice Deffune,&nbsp;Daisy Maria Fávero Salvadori","doi":"10.1002/cpz1.70370","DOIUrl":"10.1002/cpz1.70370","url":null,"abstract":"<p>For several years, scientists have focused on studying mesenchymal stem cells because of their ability for self-regeneration, their potential for differentiation into multiple lineages (e.g., osteogenic, chondrogenic, and adipogenic cells), and their low immunogenicity and remarkable capacity to modulate the immune system. The importance of these cells ranges from preserving tissue health and repairing injured tissues in their nearby areas, to their use in scientific investigation for the treatment of neurodegenerative, autoimmune, and cardiovascular diseases. Despite the availability of various tissue sources, such as bone marrow and adipose tissue, their collection and handling may not always be easily achievable. However, dental tissues, such as the pulp and periodontal ligament, are relatively accessible supplies that do not require complex or stressful interventions for the patient. Here, we provide a detailed description of each step involved in the isolation and characterization of mesenchymal stem cells from the pulp and periodontal ligament using monoclonal antibodies ensuring a high level of effectiveness. In addition, we also describe a technique to generate the cellular presenescence state. © 2026 The Author(s). <i>Current Protocols</i> published by Wiley Periodicals LLC.</p><p><b>Basic Protocol</b>: Isolation of human periodontal ligament and dental pulp stem cells</p><p><b>Support Protocol 1</b>: Characterization of human periodontal ligament and dental pulp stem cells</p><p><b>Support Protocol 2</b>: Induction of presenescence of human periodontal ligament and dental pulp stem cells</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"6 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13054634/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147629595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}