{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T09:48:34Z","timestamp":1769593714929,"version":"3.49.0"},"reference-count":34,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2024,1,23]],"date-time":"2024-01-23T00:00:00Z","timestamp":1705968000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100008658","name":"German Cancer Research Center","doi-asserted-by":"publisher","award":["NCT05836896"],"award-info":[{"award-number":["NCT05836896"]}],"id":[{"id":"10.13039\/100008658","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The search for target antigens for CAR-T cell therapy against multiple myeloma defined the B-cell maturation antigen (BCMA) as an interesting candidate. Several studies with BCMA-directed CAR-T cell therapy showed promising results. Second-generation point-of-care BCMA.CAR-T cells were manufactured to be of a GMP (good manufacturing practice) standard using the CliniMACS Prodigy\u00ae device. Cytokine release in BCMA.CAR-T cells after stimulation with BCMA positive versus negative myeloma cell lines, U266\/HL60, was assessed via intracellular staining and flow cytometry. The short-term cytotoxic potency of CAR-T cells was evaluated by chromium-51 release, while the long-term potency used co-culture (3 days\/round) at effector\/target cell ratios of 1:1 and 1:4. To evaluate the activation and exhaustion of CAR-T cells, exhaustion markers were assessed via flow cytometry. Stability was tested through a comparison of these evaluations at different timepoints: d0 as well as d + 14, d + 90 and d + 365 of cryopreservation. As results, (1) Killing efficiency of U266 cells correlated with the dose of CAR-T cells in a classical 4 h chromium-release assay. There was no significant difference after cryopreservation on different timepoints. (2) In terms of endurance of BCMA.CAR-T cell function, BCMA.CAR-T cells kept their ability to kill all tumor cells over six rounds of co-culture. (3) BCMA.CAR-T cells released high amounts of cytokines upon stimulation with tumor cells. There was no significant difference in cytokine release after cryopreservation. According to the results, BCMA.CAR-T cells manufactured under GMP conditions exerted robust and specific killing of target tumor cells with a high release of cytokines. Even after 1 year of cryopreservation, cytotoxic functions were maintained at the same level. This gives clinicians sufficient time to adjust the timepoint of BCMA.CAR-T cell application to the patient\u2019s course of the underlying disease.<\/jats:p>","DOI":"10.3390\/ijms25031394","type":"journal-article","created":{"date-parts":[[2024,1,24]],"date-time":"2024-01-24T04:54:01Z","timestamp":1706072041000},"page":"1394","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["In Vitro Functionality and Endurance of GMP-Compliant Point-of-Care BCMA.CAR-T Cells at Different Timepoints of Cryopreservation"],"prefix":"10.3390","volume":"25","author":[{"given":"Genqiao","family":"Jiang","sequence":"first","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brigitte","family":"Neuber","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angela","family":"H\u00fcckelhoven-Krauss","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Uta E.","family":"H\u00f6pken","sequence":"additional","affiliation":[{"name":"Department of Translational Tumor Immunology, Max-Delbr\u00fcck Center for Molecular Medicine (MDC), 13125 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuntian","family":"Ding","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Sedloev","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lei","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Avinoam","family":"Reichman","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Franziska","family":"Eberhardt","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martin","family":"Wermke","sequence":"additional","affiliation":[{"name":"Early Clinical Trial Unit (ECTU), Medical Clinic and Poliklinik I, Carl Gustav Carus University, 01307 Dresden, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Armin","family":"Rehm","sequence":"additional","affiliation":[{"name":"Department of Translational Tumor Immunology, Max-Delbr\u00fcck Center for Molecular Medicine (MDC), 13125 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carsten","family":"M\u00fcller-Tidow","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anita","family":"Schmitt","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1579-1509","authenticated-orcid":false,"given":"Michael","family":"Schmitt","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine V, University Clinic Heidelberg, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Kleber, M., Ntanasis-Stathopoulos, I., and Terpos, E. 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