[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-0-en-105":3,"doc-seo-444102-105":59,"doc-detail-444102-en":130},{"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":123,"head_meta":125,"extra_data":127,"updated_unix":129},105,"en","size-effect-of-stem-cell-spheroids-in-their-cryopreservation-using-low-molecular-weight-pegs-as-a-cryoprotectant","Size Effect of Stem Cell Spheroids in Their Cryopreservation Using Low-Molecular-Weight PEGs as a Cryoprotectant","","Cryopreservation is essential for long-term storage of stem cell spheroids (SCSs), but spheroid size can limit cryoprotectant action because only internalized agents function effectively. SCSs of defined size ranges were preincubated with PEGs (200, 400, or 600 Da) and frozen at −196 °C, then assessed after recovery. Recovery depended on both SCS size and PEG molecular weight, with small SCSs plus PEG200 showing superior survival, higher PEG population density, and stronger post-thaw stem-like activities. The small SCS approach also achieved >60% recovery after one month.",{"@graph":69,"@context":122},[70,84,105],{"@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/size-effect-of-stem-cell-spheroids-in-their-cryopreservation-using-low-molecular-weight-pegs-as-a-cryoprotectant/444102/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/size-effect-of-stem-cell-spheroids-in-their-cryopreservation-using-low-molecular-weight-pegs-as-a-cryoprotectant/444102.png","ImageObject",300,407,{"name":92,"@type":93},"Melati","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-03","2026-09-29",true,{"@type":102,"interactionType":103,"userInteractionCount":8},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"Why does spheroid size affect cryopreservation outcomes for stem cell spheroids?","Question",{"text":112,"@type":113},"Only cryoprotectants that penetrate into the spheroid can act effectively, so diffusion limitations in larger spheroids lead to insufficient inhibition of ice formation and osmotic stress, reducing survival and recovery of biofunctions.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"What role does PEG molecular weight play in SCS cryopreservation recovery?",{"text":117,"@type":113},"Recovery rate was significantly influenced by both SCS size and PEG molecular weight. Excellent recovery was reported for small SCSs preincubated with PEG200 solutions.",{"name":119,"@type":110,"acceptedAnswer":120},"How does the PEG200 preincubation protocol perform over longer cryopreservation durations?",{"text":121,"@type":113},"Small SCSs using the PEG200 preincubation protocol showed a cell recovery rate greater than 60% after one month of cryopreservation.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},444102,1791002809,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":39,"category_name":40,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":8,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145},962085570644,"https://ap-avatar.wpscdn.com/davatar_994ba38a5ba835b3df7d355c54d3ed8d","RESEARCH ARTICLE  \nSize Effect of Stem Cell Spheroids in Their Cryopreservation Using Low-Molecular-Weight PEGs as a Cryoprotectant  \nMadhumita Patel1,2, Solji Sung1, Brent Vernon3*, and Byeongmoon Jeong1*  \n1Department of Chemistry and Nanoscience, Graduate Program in Innovative Biomaterials Convergence, Ewha Womans University, Seoul 03760, Republic of Korea. 2Nebraska Translational Research Center (NTRC), Department of Growth and Development, College of Dentistry, University of Nebraska Medical Center, Omaha, NE 68105, USA. 3School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287-9709, USA.  \n*Address correspondence to: [brent.Vernon@asu.edu](brent.Vernon@asu.edu) (B.V.); [bjeong@ewha.ac.kr](bjeong@ewha.ac.kr) (B.J.)  \nCryopreservation is a crucial procedure to maintain quality of stem cell spheroids (SCSs) for a long period. It is affected by spheroid size because only internalized cryoprotectants can effectively play their roles within SCSs. Here, small, medium, and large SCSs with diameters of 30 to 80, 80 to 150, and 100 to 200 μm, respectively, were prepared using tonsil-derived stem cells. SCSs were preincubated in the presence of poly(ethylene glycol)s (PEGs) (10.0 wt % in a medium) with molecular weights of 200, 400, or 600 Da at 37 °C for 2 h, and then cryopreserved at −196 °C for 7 d. SCS recovery rate from cryopreservation was significantly affected by their size as well as molecular weight of PEGs; excellent recovery was observed for the small SCSs that were preincubated in PEG200 solutions. Population density of PEGs in the SCSs was 2.0 to 4.5 times higher in small SCSs than in large SCSs, which contributed to the survival of the SCSs during cryopreservation. The small SCSs recovered from cryopreservation showed innate activities of stem cells including fusion, proliferation, and differentiation much better than medium or large SCSs. The small SCSs employing the PEG200 preincubation protocol exhibited a cell recovery rate of >60% for 1 month ofcryopreservation. These findings provide valuable insights into size-dependent cryopreservation strategies for high-level complex cellular systems.  \nCitation: Patel M, Sung S, Vernon B, Jeong B. Size Effect of Stem Cell Spheroids in Their Cryopreservation Using Low-Molecular-Weight PEGs as a Cryoprotectant. Biomater. Res. 2026;30:Article 0302. [https://doi](https://doi). org/10.34133/bmr.0302  \nSubmitted 7 August 2025  \nRevised 9 November 2025  \nAccepted 6 December 2025  \nPublished 6 January 2026  \nCopyright © 2026 Madhumita Patel et al. Exclusive licensee Korean Society for Biomaterials, Republic of Korea. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License (CC BY 4 .0) .  \nIntroduction  \nCryopreservation has significantly advanced biotechnologies by enabling the long-term storage of cells, tissues, and organs while preserving their biofunctions [1] . Dimethyl sulfoxide (DMSO) is the most frequently utilized cryoprotectant. Usually, culture medium containing 10% DMSO is employed to preserve cells. DMSO replenishes intracellular water and dries the cells, and lowers osmotic stress [2,3]. However, DMSO is cytotoxic and causes harmful consequences like histone acetylation, global DNA methylation, membrane disintegration, and protein denaturation [4,5] . Additionally, stem cell differentiation may be affected by DMSO [6] . Recently, promising cryoprotectants have been extensively searched based on the cryoprotection mechanisms of ice recrystallization inhibition (IRI) and reduction of osmotic stress of cells during freezing and thawing. Antifreezing protein-mimicked polypeptides, oligonucleotides, oligopeptides, dendritic polyglycerol, graphene oxides, and synthetic polymers such as poly(vinyl alcohol), amphipathic polymers, and poly(ethylene glycol) (PEG)-polypeptides are typical examples ofcryoprotectants developed so far [3,4,7–14] . However, most of them were partially successful in a","cbCaioP9O34hQbtH","https://ap.wps.com/l/cbCaioP9O34hQbtH","pdf",14126595,13,"English","# Introduction\n## Rationale for cryopreservation and cryoprotectants\n## Limitations of DMSO and need for alternatives\n## Importance of 3D spheroids and size-dependent challenges","[{\"question\":\"Why does spheroid size affect cryopreservation outcomes for stem cell spheroids?\",\"answer\":\"Only cryoprotectants that penetrate into the spheroid can act effectively, so diffusion limitations in larger spheroids lead to insufficient inhibition of ice formation and osmotic stress, reducing survival and recovery of biofunctions.\"},{\"question\":\"What role does PEG molecular weight play in SCS cryopreservation recovery?\",\"answer\":\"Recovery rate was significantly influenced by both SCS size and PEG molecular weight. Excellent recovery was reported for small SCSs preincubated with PEG200 solutions.\"},{\"question\":\"How does the PEG200 preincubation protocol perform over longer cryopreservation durations?\",\"answer\":\"Small SCSs using the PEG200 preincubation protocol showed a cell recovery rate greater than 60% after one month of cryopreservation.\"}]","Size Effect of Stem Cell Spheroids in Their Cryopreservation Using Low-Molecular-Weight PEGs as a Cryoprotectant | PDF",1790706875,33]