{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T06:37:17Z","timestamp":1778740637108,"version":"3.51.4"},"reference-count":35,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,3,7]],"date-time":"2026-03-07T00:00:00Z","timestamp":1772841600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union from the European Regional Development Fund"},{"DOI":"10.13039\/501100011755","name":"National Center for Research and Development","doi-asserted-by":"publisher","award":["POIR.04.01.01-00-0006\/19-00"],"award-info":[{"award-number":["POIR.04.01.01-00-0006\/19-00"]}],"id":[{"id":"10.13039\/501100011755","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background: Advanced therapy medicinal products (ATMPs) require strict control of critical process parameters (CPPs) to ensure manufacturing efficiency. The relative impact of donor systemic factors, such as vitamin D status, versus technical process parameters on dental pulp-derived stem cell (DPSC) production remains unclear. Methods: In this prospective observational study, 250 adults undergoing extraction of impacted mandibular third molars were included. Dental pulp was processed under a standardized SOP using different preparation methods and enzyme conditions. Primary endpoints were serum 25(OH)D concentration and cell yield; secondary endpoints included number of passages and cryovials. Results: Mean 25(OH)D concentration was 30.1 \u00b1 14.5 ng\/mL and was higher in supplemented individuals (38.2 \u00b1 14.0 vs. 25.6 \u00b1 12.7 ng\/mL; p &lt; 0.0001) but was not associated with cell yield (\u03c1 = 0.14, p = 0.168) or passages (\u03c1 = 0.07, p = 0.406). In contrast, process parameters showed strong effects: scissor preparation resulted in a substantially higher yield than mechanical methods (median 5.00 vs. 1.00 million cells; p = 3.6 \u00d7 10\u221213), and type II collagenase was independently associated with a higher yield (+2.04 million cells; p = 0.026). The number of passages was the strongest predictor of yield (\u03b2 = 2.28 million per passage; p &lt; 10\u221226). Post-thaw viability remained high (mean 90.1% and range 81\u201398%). Conclusions: Manufacturing efficiency of DPSCs is primarily determined by critical process parameters, particularly preparation method, enzyme selection, and passage control, whereas donor vitamin D status did not significantly influence outcomes under the studied SOP. These findings highlight process standardization as the key driver of reproducible ATMP manufacturing.<\/jats:p>","DOI":"10.3390\/ph19030429","type":"journal-article","created":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T11:06:52Z","timestamp":1773054412000},"page":"429","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["From Extraction to Cryobanking: Which Critical Process Parameters Genuinely Improve DPSC Production?"],"prefix":"10.3390","volume":"19","author":[{"given":"Tomasz","family":"Gedrange","sequence":"first","affiliation":[{"name":"Department of Dental Surgery, Wroclaw Medical University, Krakowska 26, 50-425 Wroclaw, Poland"},{"name":"Institute of Pharmacology and Toxicology, Jena University Hospital, Friedrich Schiller University Jena, Drackendorfer Stra\u00dfe 1, D-07747 Jena, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1404-2274","authenticated-orcid":false,"given":"Benita","family":"Wiatrak","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Faculty of Medicine, Wroclaw Medical University, Mikulicza-Radeckiego 2, 50-345 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8742-0790","authenticated-orcid":false,"given":"Tomasz","family":"G\u0119barowski","sequence":"additional","affiliation":[{"name":"Department of Biostructure and Animal Physiology, Wroclaw University of Environmental and Life Sciences, Ko\u017cuchowska 1\/3, 51-631 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ewa","family":"Barg","sequence":"additional","affiliation":[{"name":"Department of Basic Medical Sciences and Immunology, Faculty of Pharmacy, Wroclaw Medical University, Borowska 211, 50-556 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7736-1427","authenticated-orcid":false,"given":"\u0141ucja","family":"Cwynar-Zaj\u0105c","sequence":"additional","affiliation":[{"name":"Department of Basic Medical Sciences and Immunology, Faculty of Pharmacy, Wroclaw Medical University, Borowska 211, 50-556 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2670-0569","authenticated-orcid":false,"given":"Katarzyna","family":"G\u0119bczak","sequence":"additional","affiliation":[{"name":"Department of Basic Medical Sciences and Immunology, Faculty of Pharmacy, Wroclaw Medical University, Borowska 211, 50-556 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8084-3686","authenticated-orcid":false,"given":"Helena","family":"Moreira","sequence":"additional","affiliation":[{"name":"Department of Basic Medical Sciences and Immunology, Faculty of Pharmacy, Wroclaw Medical University, Borowska 211, 50-556 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aneta","family":"Cie\u015bla-Niechwiadowicz","sequence":"additional","affiliation":[{"name":"Department of Biostructure and Animal Physiology, Wroclaw University of Environmental and Life Sciences, Ko\u017cuchowska 1\/3, 51-631 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2353-3198","authenticated-orcid":false,"given":"Jakub","family":"Hadzik","sequence":"additional","affiliation":[{"name":"Department of Dental Surgery, Wroclaw Medical University, Krakowska 26, 50-425 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0141-7310","authenticated-orcid":false,"given":"Amelie","family":"Lupp","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology and Toxicology, Jena University Hospital, Friedrich Schiller University Jena, Drackendorfer Stra\u00dfe 1, D-07747 Jena, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sophie","family":"Gedrange","sequence":"additional","affiliation":[{"name":"Student Research Group, Department of Dental Surgery, Faculty of Dentistry, Wroclaw Medical University, Krakowska 26, 50-425 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8943-0549","authenticated-orcid":false,"given":"Marzena","family":"Dominiak","sequence":"additional","affiliation":[{"name":"Department of Dental Surgery, Wroclaw Medical University, Krakowska 26, 50-425 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,7]]},"reference":[{"key":"#cr-split#-ref_1.1","unstructured":"European Union (2007). 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