{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T07:03:00Z","timestamp":1771052580742,"version":"3.50.1"},"reference-count":43,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:00:00Z","timestamp":1764288000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:00:00Z","timestamp":1764288000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2026,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Studies on cell cryopreservation have been limited by the complexity of the freezing process and challenges on controlling ice formation, managing cooling rates, and optimizing cryoprotectant concentrations. The objective of this study is to evaluate the impact of bottom\u2010up and conventional radial freezing on the viability of mammalian cells, using mouse hybridoma cells and human umbilical cord blood (hUCB) derived mononuclear cells (MNCs) as models. UCB\u2010derived MNCs were selected for this study because these contain hematopoietic stem and progenitor cells (HSPCs), which hold significant clinical relevance. The study combines experimental assays, including cell viability assays and flow cytometry characterization, with Computational Fluid Dynamic (CFD) simulations. A bottom\u2010up freezing geometry sustained high cell viability, even at dimethyl sulfoxide (DMSO) concentrations below 5% (v\/v), whereas conventional radial freezing led to lower cell viability when DMSO is used below such concentrations threshold. This observation is particularly relevant for cell\u2010based therapies. CFD simulations for conventional radial freezing elucidated that for such method the ice formed at the top of the vial is of high porosity for media with 10% (v\/v) DMSO, but of low porosity for lower DMSO concentrations. The simulations show that the latter conditions can result in an increase in shear stress on cells, by up to an order of magnitude. Overall, this study provides a rational for 10% (v\/v) DMSO being the optimal reported concentration for conventional freezing methods, as a result of poor control of ice growth direction and higher mechanical stresses at lower DMSO concentrations. Experimental results show that bottom\u2010up freezing method, using only 2.5% (v\/v) DMSO, allow to reach cell viabilities as high as the ones obtained with conventional radial freezing protocols at 10% (v\/v) DMSO. In addition, bottom\u2010up freezing method with 2.5% DMSO preserves the clonogenic potential of HSPCs within hUCB\u2010derived MNCs comparably to conventional radial freezing protocol with 10% DMSO. Importantly, the results support the recommendation to use cell cryopreservation strategies, such as bottom\u2010up freezing, that enable the use of lower DMSO concentrations by controlling the direction of heat transfer.<\/jats:p>","DOI":"10.1002\/bit.70116","type":"journal-article","created":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T13:25:01Z","timestamp":1764336301000},"page":"582-597","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Bottom\u2010Up Ice Growth Geometry Attenuates Shear Stress and Improves the Cryopreservation of Hematopoietic Stem\/Progenitor Cells Under Low DMSO Concentrations"],"prefix":"10.1002","volume":"123","author":[{"given":"Rafaela Ouro","family":"Neves","sequence":"first","affiliation":[{"name":"SmartFreez, Ed. 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