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More than 30 adipogenic condition combinations were screened with commercial cells, then assessed by microscopy, Oil Red O staining, UCP1 expression via RT-qPCR, and Western blot. Rosiglitazone outperformed indomethacin; extending induction from 4 to 8 days and sustaining dexamethasone improved efficiency. Results inhibited differentiation above 5% serum, while optimal conditions were 5 µM rosiglitazone and 20 µg/mL insulin, yielding donor-dependent efficiency.",{"@graph":14,"@context":77},[15,34,56],{"@type":16,"itemListElement":17},"BreadcrumbList",[18,23,27,31],{"item":19,"name":20,"@type":21,"position":22},"https://docshare.wps.com","Home","ListItem",1,{"item":24,"name":25,"@type":21,"position":26},"https://docshare.wps.com/document/","Document",2,{"item":28,"name":29,"@type":21,"position":30},"https://docshare.wps.com/document/healthcare/","Healthcare",3,{"item":32,"name":10,"@type":21,"position":33},"https://docshare.wps.com/document/toward-efficient-beige-adipogenesis-protocol-optimization-using-adipose-derived-stem-cells/465206/",4,{"url":32,"name":10,"@type":35,"image":36,"author":41,"headline":10,"publisher":44,"fileFormat":47,"inLanguage":8,"description":12,"dateModified":48,"datePublished":49,"encodingFormat":47,"isAccessibleForFree":50,"interactionStatistic":51},"DigitalDocument",{"url":37,"@type":38,"width":39,"height":40},"https://docshare.wps.com/thumbnails/toward-efficient-beige-adipogenesis-protocol-optimization-using-adipose-derived-stem-cells/465206.png","ImageObject",300,407,{"name":42,"@type":43},"Genevieve","Person",{"url":19,"name":45,"@type":46},"DocShare","Organization","application/pdf","2026-10-31","2026-09-30",true,{"@type":52,"interactionType":53,"userInteractionCount":55},"InteractionCounter",{"@type":54},"ViewAction",7,{"@type":57,"mainEntity":58},"FAQPage",[59,65,69,73],{"name":60,"@type":61,"acceptedAnswer":62},"What is the main goal of this study on beige adipogenesis?","Question",{"text":63,"@type":64},"To optimize and validate an in vitro protocol that differentiates adipose-derived stem cells (ADSCs) into beige adipocytes, supporting studies of beige fat biology and BAT activation mechanisms.","Answer",{"name":66,"@type":61,"acceptedAnswer":67},"How was adipogenic differentiation measured in the optimized protocol?",{"text":68,"@type":64},"Differentiation was assessed by microscopy, Oil Red O staining, and evaluation of uncoupling protein 1 (UCP1) expression using RT-qPCR and Western blot.",{"name":70,"@type":61,"acceptedAnswer":71},"Which conditions improved differentiation efficiency the most?",{"text":72,"@type":64},"Rosiglitazone was more effective than indomethacin; extending the induction phase from 4 to 8 days and maintaining dexamethasone throughout culture markedly increased differentiation efficiency.",{"name":74,"@type":61,"acceptedAnswer":75},"What donor-related limitation was observed after optimization?",{"text":76,"@type":64},"The optimized protocol successfully induced beige adipogenesis in ADSCs from eight donors, but differentiation efficiency varied considerably, attributed to individual donor variability.","https://schema.org",{"og:url":32,"og:type":79,"og:title":10,"og:site_name":45,"og:description":12},"article",{"robots":81,"canonical":32},"index,follow",{"doc_id":83,"site_id":7},465206,1790813867,{"code":4,"msg":86,"data":87},"success",[88,92,96,100,105,110,113,118,123,126,130],{"id":22,"doc_module":4,"doc_module_name":25,"category_name":89,"show_sort_weight":90,"slug":91},"Story & Novel",90,"story-novel",{"id":26,"doc_module":4,"doc_module_name":25,"category_name":93,"show_sort_weight":94,"slug":95},"Literature",80,"literature",{"id":33,"doc_module":4,"doc_module_name":25,"category_name":97,"show_sort_weight":98,"slug":99},"Exam",70,"exam",{"id":101,"doc_module":4,"doc_module_name":25,"category_name":102,"show_sort_weight":103,"slug":104},5,"Comic",60,"comic",{"id":106,"doc_module":4,"doc_module_name":25,"category_name":107,"show_sort_weight":108,"slug":109},6,"Technology",50,"technology",{"id":55,"doc_module":4,"doc_module_name":25,"category_name":29,"show_sort_weight":111,"slug":112},40,"healthcare",{"id":114,"doc_module":4,"doc_module_name":25,"category_name":115,"show_sort_weight":116,"slug":117},8,"Research & Report",30,"research-report",{"id":119,"doc_module":4,"doc_module_name":25,"category_name":120,"show_sort_weight":121,"slug":122},9,"Religion & Spirituality",20,"religion-spirituality",{"id":121,"doc_module":4,"doc_module_name":25,"category_name":124,"show_sort_weight":121,"slug":125},"World Cup","world-cup",{"id":127,"doc_module":4,"doc_module_name":25,"category_name":128,"show_sort_weight":127,"slug":129},10,"Lifestyle","lifestyle",{"id":131,"doc_module":4,"doc_module_name":25,"category_name":132,"show_sort_weight":101,"slug":133},19,"General","general",{"code":4,"msg":86,"data":135},{"doc_id":83,"user_id":136,"nickname":42,"user_avatar":137,"doc_module":4,"category_id":55,"category_name":29,"doc_title":10,"doc_description":12,"doc_content":138,"file_id":139,"file_url":140,"file_type":141,"file_size":142,"view_count":55,"is_deleted":4,"is_public":22,"is_downloadable":22,"audit_status":22,"page_count":143,"language":144,"language_code":8,"site_id":7,"html_lang":8,"table_of_contents":145,"faqs":146,"seo_title":147,"seo_description":12,"update_tm":148,"read_time":103,"preview_page_count":4},1374391974585,"https://ap-avatar.wpscdn.com/davatar_276721f389ce27ea32af1340a28f341c","Article  \nToward Efficient Beige Adipogenesis: Protocol Optimization Using Adipose-Derived Stem Cells  \nKlaudia Simka-Lampa 1, *, Agnieszka Kosowska 1, Wojciech Garczorz 1, Małgorzata Kimsa-Furdzik 1, Grzegorz Wystrychowski 2, Celina Kruszniewska-Rajs 3, Małgorzata Muc-Wierzgo ´n 4 and Tomasz Francuz 1  \nAcademic Editor: Alexander E. Kalyuzhny, Silvia Canaiderand Federica Facchin  \nReceived: 6 October 2025  \nRevised: 3 December 2025  \nAccepted: 20 December 2025  \nPublished: 28 December 2025  \nCopyright: © 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license.  \n1 Department of Biochemistry, Faculty of Medical Sciences in Katowice, Medical University of Silesia, Medyków 18, 40-752 Katowice, Poland  \n2 Department of Nephrology, 4th Provincial Hospital, 41-902 Bytom, Poland  \n3 Department of Molecular Biology, Faculty of Pharmaceutical Sciences in Sosnowiec, Medical University of Silesia, Jedno´sci 8, 41-200 Sosnowiec, Poland  \n4 Department of Internal Diseases Propaedeutics and Emergency Medicine, Faculty of Public Health in Bytom, Medical University of Silesia, Piekarska 18, 44-902 Bytom, Poland  \n* Correspondence: [ksimka@sum.edu.pl](ksimka@sum.edu.pl); Tel.: +48-(32)-25-25-088  \nAbstract  \nBrown adipose tissue (BAT) has emerged as a promising therapeutic target for metabolic disorders such as type 2 diabetes and obesity. To advance research on BAT activation and elucidate the mechanisms underlying adipogenesis, it is crucial to develop a reliable in vitro model. This study aimed to optimize the differentiation of adipose-derived stem cells (ADSCs) into beige adipocytes and to validate the protocol using primary human ADSCs obtained from eight donors. Protocol optimization was first performed with commercial ADSCs, testing more than 30 combinations of adipogenic conditions. Differentiation was assessed by microscopy, Oil Red O staining, and uncoupling protein 1 (UCP1) expression via reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot. Among the key adipogenic factors, rosiglitazone proved more effective than indomethacin. Extending the induction phase from 4 to 8 days and maintaining dexamethasone throughout the culture markedly enhanced differentiation efficiency. Serum concentration above 5% was inhibitory, while optimal conditions were identified as 5 µM rosiglitazone and 20 µg/mL insulin. The optimized protocol successfully induced beige adipogenesis in ADSCs from eight independent donors, though efficiency varied considerably which could be attributed to individual donor variability. These findings provide a robust in vitro model for studying beige fat biology and highlight the relevance of personalized approaches in metabolic research.  \nKeywords: adipogenesis; beige adipocytes; brown adipose tissue; adipose-derived stem cells; differentiation; uncoupling protein 1 (UCP1); thermogenin  \n1. Introduction  \nBrown adipose tissue (BAT) plays a pivotal role in thermoregulation in newborns and young children by mediating non-shivering adaptive thermogenesis through the expression of uncoupling protein 1 (UCP1, thermogenin), which is responsible for uncoupling oxidative phosphorylation in mitochondria. BAT decreases with age and gradually transforms into white adipose tissue (WAT) [1] . Due to advances in modern research methods, BAT has been confirmed to persist in various adult tissues [2,3] and shown to play a key role in metabolic regulation [3,4], overturning the earlier belief that it regressed completely  \nwith age. Studies in animal models have shown that BAT transplantation restores euglycemia, lowers levels of pro-inflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α), and reverses clinical symptoms of diabetes [5] . Moreover, it reduces body weight by increasing energy expenditure without altering food intake [6]","cbCaiqx84UMtFQ0b","https://ap.wps.com/l/cbCaiqx84UMtFQ0b","pdf",9271138,24,"English","# Abstract\n# Introduction","[{\"question\":\"What is the main goal of this study on beige adipogenesis?\",\"answer\":\"To optimize and validate an in vitro protocol that differentiates adipose-derived stem cells (ADSCs) into beige adipocytes, supporting studies of beige fat biology and BAT activation mechanisms.\"},{\"question\":\"How was adipogenic differentiation measured in the optimized protocol?\",\"answer\":\"Differentiation was assessed by microscopy, Oil Red O staining, and evaluation of uncoupling protein 1 (UCP1) expression using RT-qPCR and Western blot.\"},{\"question\":\"Which conditions improved differentiation efficiency the most?\",\"answer\":\"Rosiglitazone was more effective than indomethacin; extending the induction phase from 4 to 8 days and maintaining dexamethasone throughout culture markedly increased differentiation efficiency.\"},{\"question\":\"What donor-related limitation was observed after optimization?\",\"answer\":\"The optimized protocol successfully induced beige adipogenesis in ADSCs from eight donors, but differentiation efficiency varied considerably, attributed to individual donor variability.\"}]","Toward Efficient Beige Adipogenesis - Protocol Optimization Using Adipose-Derived Stem Cells | PDF",1790769312]