[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-0-en-105":3,"doc-seo-431307-105":59,"doc-detail-431307-en":129},{"code":4,"msg":5,"data":6},0,"success",[7,13,18,23,28,33,38,43,48,51,55],{"id":8,"doc_module":4,"doc_module_name":9,"category_name":10,"show_sort_weight":11,"slug":12},1,"Document","Story & Novel",90,"story-novel",{"id":14,"doc_module":4,"doc_module_name":9,"category_name":15,"show_sort_weight":16,"slug":17},2,"Literature",80,"literature",{"id":19,"doc_module":4,"doc_module_name":9,"category_name":20,"show_sort_weight":21,"slug":22},4,"Exam",70,"exam",{"id":24,"doc_module":4,"doc_module_name":9,"category_name":25,"show_sort_weight":26,"slug":27},5,"Comic",60,"comic",{"id":29,"doc_module":4,"doc_module_name":9,"category_name":30,"show_sort_weight":31,"slug":32},6,"Technology",50,"technology",{"id":34,"doc_module":4,"doc_module_name":9,"category_name":35,"show_sort_weight":36,"slug":37},7,"Healthcare",40,"healthcare",{"id":39,"doc_module":4,"doc_module_name":9,"category_name":40,"show_sort_weight":41,"slug":42},8,"Research & Report",30,"research-report",{"id":44,"doc_module":4,"doc_module_name":9,"category_name":45,"show_sort_weight":46,"slug":47},9,"Religion & Spirituality",20,"religion-spirituality",{"id":46,"doc_module":4,"doc_module_name":9,"category_name":49,"show_sort_weight":46,"slug":50},"World Cup","world-cup",{"id":52,"doc_module":4,"doc_module_name":9,"category_name":53,"show_sort_weight":52,"slug":54},10,"Lifestyle","lifestyle",{"id":56,"doc_module":4,"doc_module_name":9,"category_name":57,"show_sort_weight":24,"slug":58},19,"General","general",{"code":4,"msg":60,"data":61},"ok",{"site_id":62,"language":63,"slug":64,"title":65,"keywords":66,"description":67,"schema_data":68,"social_meta":122,"head_meta":124,"extra_data":126,"updated_unix":128},105,"en","decellularized-lymph-node-sections-with-preserved-extracellular-matrix-for-stromal-cell-culture","Decellularized lymph node sections with preserved extracellular matrix for stromal cell culture","","Decellularized lymph node extracellular matrix scaffolds are used to study how the native lymph node microenvironment regulates stromal and immune cell behavior, overcoming limits of existing whole-organ decellularization models and animal-dependent approaches. The protocol generates thin 200-μm lymph node slices using vibratome sectioning combined with detergent decellularization (0.1% SDS and 1% Triton-X). Preserved collagen and glycosaminoglycans, immunofluorescence-detected ECM proteins, 21-day fibroblastic reticular cell culture, co-culture with T cells, and high-resolution imaging/flow cytometry reveal altered gp38 and PDGFRα expression versus 2D culture.",{"@graph":69,"@context":121},[70,84,104],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":40,"@type":76,"position":81},"https://docshare.wps.com/document/research-report/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/decellularized-lymph-node-sections-with-preserved-extracellular-matrix-for-stromal-cell-culture/431307/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":98,"encodingFormat":97,"isAccessibleForFree":99,"interactionStatistic":100},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/decellularized-lymph-node-sections-with-preserved-extracellular-matrix-for-stromal-cell-culture/431307.png","ImageObject",300,407,{"name":92,"@type":93},"Fahsai","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-09-29",true,{"@type":101,"interactionType":102,"userInteractionCount":4},"InteractionCounter",{"@type":103},"ViewAction",{"@type":105,"mainEntity":106},"FAQPage",[107,113,117],{"name":108,"@type":109,"acceptedAnswer":110},"Why decellularize lymph node tissue for stromal cell culture studies?","Question",{"text":111,"@type":112},"Decellularized lymph node scaffolds preserve the native ECM structure, composition, and biochemical cues while removing cells that confound mechanistic experiments. This enables in vitro analysis of stromal–ECM interactions and their impact on cellular behavior.","Answer",{"name":114,"@type":109,"acceptedAnswer":115},"How does the presented protocol generate decellularized lymph node slices?",{"text":116,"@type":112},"It combines vibratome sectioning into 200-μm slices with detergent decellularization using 0.1% SDS and 1% Triton-X to produce thin lymph node sections suitable for culture and analysis.",{"name":118,"@type":109,"acceptedAnswer":119},"What functional outcomes were observed after decellularization?",{"text":120,"@type":112},"Decellularized sections sustained 21-day FRC culture, supported FRC–T cell co-culture, and enabled high-resolution imaging and flow cytometric analyses, including altered gp38 and PDGFRα expression compared with 2D culture.","https://schema.org",{"og:url":83,"og:type":123,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":125,"canonical":83},"index,follow",{"doc_id":127,"site_id":62},431307,1790655111,{"code":4,"msg":5,"data":130},{"doc_id":127,"user_id":131,"nickname":92,"user_avatar":132,"doc_module":4,"category_id":39,"category_name":40,"doc_title":65,"doc_description":67,"doc_content":133,"file_id":134,"file_url":135,"file_type":136,"file_size":137,"view_count":4,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":138,"language":139,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":140,"faqs":141,"seo_title":142,"seo_description":67,"update_tm":128,"read_time":36},549768702563,"https://ap-avatar.wpscdn.com/avatar/8000c4aa63b76e948b?x-image-process=image/resize,m_fixed,w_180,h_180&k=1786536092046926083","[www. nature.com/scientificreports](www. nature.com/scientificreports)  \nOPEN  \nDecellularized lymph node sections with preserved extracellular matrix for stromal cell culture  \nEstefania Esparza1,2,4, Leonor N. Teles1,2,4􀀍, Alisa Fedotova2, Noa Dehaseth1,  \nMira Sayegh2,3, Ana V. Hernandez2, Lucy Y. Ho2, Noel M. Ziebarth1 & Alice A. Tomei1,2,3􀀍  \nThe lymph node (LN) extracellular matrix (ECM) is produced by stromal cells like fibroblastic reticular cells (FRCs) and supports adaptive immunity by guiding immune cell interactions. Disruption of this ECM in cancer and chronic inflammation has been shown to promote disease progression. While interactions between cells and the LN ECM are critical for immunity, they remain difficult to study due to limitations in current models and reliance on animal studies. To address this, LNs could bedecellularized to generate cell-free scaffolds that are subsequently reseeded with cells to study how the native LN microenvironment influences cellular behavior. Existing whole-organ decellularization methods preserve ECM features but yield dense scaffolds that restrict uniform cell seeding, limit nutrient diffusion, and hinder imaging analyses. Here, we present a protocol that combines vibratome sectioning (200-μm slices) with detergent decellularization (0.1% SDS and 1% Triton-X) to generate thin LN slices from mouse and human tissues. Decellularized LNs had comparable collagen and GAG concentrations to native tissue, and immunofluorescence staining showed the presence of other ECM proteins. Decellularized sections sustained 21-day FRC culture, enabled FRC-T cell co-culture, and supported high-resolution imaging and flow cytometric analyses, revealing altered gp38 and PDGFRα expression in FRCs relative to 2D culture.  \nKeywords Decellularization, Extracellular matrix, Lymphoid organs, 3D cell culture, Fibroblastic reticular cells  \nLymph nodes (LNs) are essential organs of the immune system where adaptive immune responses are initiated and regulated1. Within the LN paracortex, fibroblastic reticular cells (FRCs), a specialized stromal cell population, produce a network of extracellular matrix (ECM) fibers that guide immune cell migration, activation and interactions2–5. This highly organized ECM microenvironment is composed of collagens, glycosaminoglycans (GAGs), proteoglycans, and glycoproteins6,7. Collagen provides structural support and tensile strength8, while glycoproteins like laminin and fibronectin form basement membranes that facilitate cell adhesion and signaling through integrin-binding domains5,9. GAGs such as hyaluronic acid (HA) support cell trafficking and motility7. Notably, the molecular weight of HA influences immune cell activation: high molecular weight HA suppresses immune responses, while fragmented, low molecular weight HA promotes inflammation10, 11. Proteoglycans including perlecan and decorin contribute to the ECM’s mechanical properties, regulate growth factor availability and influence cell proliferation and differentiation7, 10, 12, 13. Together, these ECM components create a highly organized microenvironment that is critical for LN architecture and immunity.  \nGiven the ECM’s role in immune homeostasis, its components are increasingly recognized as therapeutic targets across diseases14, 15. In cancer, changes in ECM architecture drive tumor progression and indicate disease state16. Similarly, in autoimmune diseases like type 1 diabetes, collagen and laminin remodeling has been linked to immune cell infiltration and tissue destruction11. Multiple groups have shown that pancreatic LNs from individuals with recent-onset type 1 diabetes display altered ECM composition, including reduced germinal centers and fragmented HA, which promote immune activation and accelerate disease progression17, 18. Our lab has also previously demonstrated that FRC networks in the pancreatic and skin-draining LNs of non-obese diabetic mice (NOD) display larger reticular pores than non-obese ","cbCailaAFnZj8DwH","https://ap.wps.com/l/cbCailaAFnZj8DwH","pdf",8204406,16,"English","# Background\n## Role of lymph node ECM in immunity\n## Disease links and need for improved models\n# Methods and Protocol\n## Vibratome sectioning and detergent decellularization\n# Results\n## ECM preservation and staining\n## Cell culture outcomes: FRC monoculture and FRC–T co-culture\n## Imaging and flow cytometric readouts","[{\"question\":\"Why decellularize lymph node tissue for stromal cell culture studies?\",\"answer\":\"Decellularized lymph node scaffolds preserve the native ECM structure, composition, and biochemical cues while removing cells that confound mechanistic experiments. This enables in vitro analysis of stromal–ECM interactions and their impact on cellular behavior.\"},{\"question\":\"How does the presented protocol generate decellularized lymph node slices?\",\"answer\":\"It combines vibratome sectioning into 200-μm slices with detergent decellularization using 0.1% SDS and 1% Triton-X to produce thin lymph node sections suitable for culture and analysis.\"},{\"question\":\"What functional outcomes were observed after decellularization?\",\"answer\":\"Decellularized sections sustained 21-day FRC culture, supported FRC–T cell co-culture, and enabled high-resolution imaging and flow cytometric analyses, including altered gp38 and PDGFRα expression compared with 2D culture.\"}]","Decellularized lymph node sections with preserved extracellular matrix for stromal cell culture | PDF"]