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Herein, we report a tardigrade-inspired, glycerol-free strategy for RBC cryopreservation based on a conserved CAHS-derived peptide motif. Systematic sequence analysis identified a minimal CAHS-motif that possesses an intrinsic helical propensity and adopts a stabilized amphipathic \u03b1-helical conformation under dehydration-mimicking conditions. Under the 4 \u00b0C loading condition, CAHS-motif and trehalose co-incubation was associated with increased membrane fluidity and increased intracellular trehalose accumulation while maintaining low pre-freeze hemolysis. In parallel, the CAHS-motif acts synergistically with trehalose to suppress ice-associated damage during freezing and thawing. Under the combined effect, RBCs cryopreserved with the CAHS-motif and trehalose formulation achieved a post-thaw recovery of 89.0 \u00b1 0.6% and excellent blood compatibility (99.0 \u00b1 0.7%), outperforming the conventional glycerol-based method. Post-thaw RBCs retained normal morphology, volume, and key functional activities in vitro and effectively corrected anemia in an APH-induced hemolytic anemia mouse model without detectable pro-inflammatory responses. These results establish a clinically relevant, bioinspired platform for high-recovery RBC cryopreservation and highlight the translational potential of extremophile-derived motifs in transfusion medicine.<\/jats:p>","DOI":"10.1021\/acsami.6c12445","type":"journal-article","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T01:46:54Z","timestamp":1790214414000},"page":"48828-48843","source":"Crossref","is-referenced-by-count":0,"title":["A Tardigrade-Derived CAHS-Motif Enables Glycerol-Free Red Blood Cell Cryopreservation with Enhanced Low-Temperature Trehalose Loading and Synergistic Suppression of Ice Damage"],"prefix":"10.1021","volume":"18","author":[{"given":"Tianwen","family":"Xi","sequence":"first","affiliation":[{"name":"Northwestern Polytechnical University , , , ,","place":["Xi\u2019an, Shaanxi, China, 710072"]},{"name":"Dongguan Sanhang Innovation Institute , , ,","place":["Dongguan, Guangdong, China, 523808"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiyi","family":"Ding","sequence":"additional","affiliation":[{"name":"Northwestern Polytechnical University , , , ,","place":["Xi\u2019an, Shaanxi, China, 710072"]},{"name":"Dongguan Sanhang Innovation Institute , , ,","place":["Dongguan, Guangdong, China, 523808"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruichen","family":"Wang","sequence":"additional","affiliation":[{"name":"Northwestern Polytechnical University , , , ,","place":["Xi\u2019an, Shaanxi, China, 710072"]},{"name":"Tsinghua University , , ,","place":["Beijing, China, 100084"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujuan","family":"Ren","sequence":"additional","affiliation":[{"name":"Northwestern Polytechnical University , , , ,","place":["Xi\u2019an, Shaanxi, China, 710072"]},{"name":"Dongguan Sanhang Innovation Institute , , ,","place":["Dongguan, Guangdong, China, 523808"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuanlin","family":"Qu","sequence":"additional","affiliation":[{"name":"Northwestern Polytechnical University , , , ,","place":["Xi\u2019an, Shaanxi, China, 710072"]},{"name":"Dongguan Sanhang Innovation Institute , , ,","place":["Dongguan, Guangdong, China, 523808"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meilian","family":"Chen","sequence":"additional","affiliation":[{"name":"Northwestern Polytechnical University , , , ,","place":["Xi\u2019an, Shaanxi, China, 710072"]},{"name":"Dongguan Sanhang Innovation Institute , , ,","place":["Dongguan, Guangdong, China, 523808"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingli","family":"Chu","sequence":"additional","affiliation":[{"name":"Northwestern Polytechnical University , , , ,","place":["Xi\u2019an, Shaanxi, China, 710072"]},{"name":"Dongguan Sanhang Innovation Institute , , ,","place":["Dongguan, Guangdong, China, 523808"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yijia","family":"Chen","sequence":"additional","affiliation":[{"name":"Sichuan Agricultural University , , , ,","place":["Chengdu, Sichuan, China, 611130"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7567-3048","authenticated-orcid":true,"given":"Hui","family":"Yang","sequence":"additional","affiliation":[{"name":"Northwestern Polytechnical University , , , ,","place":["Xi\u2019an, Shaanxi, China, 710072"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3188-9557","authenticated-orcid":true,"given":"Leming","family":"Sun","sequence":"additional","affiliation":[{"name":"Northwestern Polytechnical University , , , ,","place":["Xi\u2019an, Shaanxi, China, 710072"]},{"name":"Dongguan Sanhang Innovation Institute , , ,","place":["Dongguan, Guangdong, China, 523808"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,4]]},"reference":[{"key":"2026092321464402300_cit1","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1007\/978-1-4939-8994-2_22","article-title":"Cryopreservation of Red Blood Cells","volume-title":"Pre-Clinical Models: Techniques and Protocols","author":"Al-Otaibi","year":"2019"},{"issue":"4","key":"2026092321464402300_cit2","doi-asserted-by":"publisher","first-page":"719","DOI":"10.1002\/bit.22435","article-title":"Do physical forces contribute to cryodamage?","volume":"104","author":"Saragusty","year":"2009","journal-title":"Biotechnol. 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