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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 \u03bcm, 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 \u00b0C for 2 h, and then cryopreserved at \u2212196 \u00b0C 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 &gt;60% for 1 month of cryopreservation. These findings provide valuable insights into size-dependent cryopreservation strategies for high-level complex cellular systems.<\/jats:p>","DOI":"10.34133\/bmr.0302","type":"journal-article","created":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T21:37:29Z","timestamp":1765229849000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark_01","source":"Crossref","is-referenced-by-count":4,"title":["Size Effect of Stem Cell Spheroids in Their Cryopreservation Using Low-Molecular-Weight PEGs as a Cryoprotectant"],"prefix":"10.34133","volume":"30","author":[{"given":"Madhumita","family":"Patel","sequence":"first","affiliation":[{"name":"Department of Chemistry and Nanoscience, Graduate Program in Innovative Biomaterials Convergence, \rEwha Womans University, Seoul 03760, Republic of Korea."},{"name":"Nebraska Translational Research Center (NTRC), Department of Growth and Development, College of Dentistry, \rUniversity of Nebraska Medical Center, Omaha, NE 68105, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Solji","family":"Sung","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Nanoscience, Graduate Program in Innovative Biomaterials Convergence, \rEwha Womans University, Seoul 03760, Republic of Korea."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brent","family":"Vernon","sequence":"additional","affiliation":[{"name":"School of Biological and Health Systems Engineering, \rArizona State University, Tempe, AZ 85287-9709, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Byeongmoon","family":"Jeong","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Nanoscience, Graduate Program in Innovative Biomaterials Convergence, \rEwha Womans University, Seoul 03760, Republic of Korea."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","published-online":{"date-parts":[[2026,1,6]]},"reference":[{"issue":"40","key":"e_1_3_3_2_2","doi-asserted-by":"crossref","first-page":"2303373","DOI":"10.1002\/adfm.202303373","article-title":"Cryopreservation in the era of cell therapy: Revisiting fundamental concepts to enable future technologies","volume":"33","author":"Bai B","year":"2023","unstructured":"Bai B, Xue C, Wen Y, Lim J, Le Z, Shou Y, Shin S, Tay A. 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