{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T22:23:42Z","timestamp":1769725422140,"version":"3.49.0"},"reference-count":49,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T00:00:00Z","timestamp":1768521600000},"content-version":"vor","delay-in-days":15,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J of Cellular Biochemistry"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Lysosomal function can be affected by components in cell culture. This in turn may influence cellular metabolism and, consequently, research and diagnostics outcomes. One such component is the commonly used pH buffer 4\u2010(2\u2010hydroxyethyl)\u20101\u2010piperazineethanesulfonic acid (HEPES). HEPES specifically impacts the trafficking of the lysosomal enzyme glucocerebrosidase, which is deficient in Gaucher disease (GD). Understanding how HEPES affects cellular models of GD is essential, since glucocerebrosidase is central to diagnostic testing and the investigation of GD pathophysiology. Therefore, we examined the broader effects of HEPES on cultured fibroblasts from individuals with GD and healthy controls. We cultured dermal fibroblasts of eight adults with GD and seven healthy age\u2010 and sex\u2010matched controls. The cells were cultured in two culture media, Ham's F10 and DMEM, both with and without HEPES. We assessed glucocerebrosidase enzyme activity and sphingolipid concentrations using a quantitative UPLC\u2010MS\/MS method. Additionally, we conducted multi\u2010omics analyses, consisting of lipidomics, metabolomics and proteomics, to explore the broader impact of HEPES in cell culture on fibroblasts. Glucocerebrosidase activity in cell lysates increased after HEPES exposure in both GD and control fibroblasts, to an extent that may influence diagnostic outcomes. In GD fibroblasts, substrate accumulation was absent and not altered by HEPES exposure. GD fibroblasts exhibited a multi\u2010omics profile largely overlapping with healthy controls and lacking the typical pathological features associated with GD in other cell types, such as mitochondrial dysfunction, dysregulated autophagy, disruption of intracellular calcium homeostasis, ER stress and chronic oxidative stress. In addition, the multi\u2010omics profile was altered by HEPES, however in a non\u2010specific manner. In conclusion, HEPES influences fibroblasts in culture, both from healthy controls and from patients with GD. Furthermore, GD fibroblasts lack a specific disease\u2010related profile. This renders cultured fibroblasts unsuitable for studying pathophysiological processes in GD. Culturing GD fibroblasts with HEPES may compromise the reliability of diagnostics.<\/jats:p>","DOI":"10.1002\/jcb.70080","type":"journal-article","created":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T09:00:29Z","timestamp":1768554029000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["HEPES in Cell Culture Alters the Multi\u2010Omics Profile Exhibited by Gaucher Disease Fibroblasts"],"prefix":"10.1002","volume":"127","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2787-2549","authenticated-orcid":false,"given":"Eleonore M.","family":"Corazolla","sequence":"first","affiliation":[{"name":"Department of Endocrinology and Metabolism Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"},{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"},{"name":"Amsterdam Gastroenterology Endocrinology and Metabolism Institute Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0242-9113","authenticated-orcid":false,"given":"Bauke V.","family":"Schomakers","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"},{"name":"Core Facility Metabolomics Amsterdam UMC Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria M.","family":"Tr\u0119towicz","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"},{"name":"Amsterdam Gastroenterology Endocrinology and Metabolism Institute Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jill","family":"Hermans","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"},{"name":"Core Facility Metabolomics Amsterdam UMC Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4916-2866","authenticated-orcid":false,"given":"Michel","family":"van Weeghel","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"},{"name":"Core Facility Metabolomics Amsterdam UMC Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fr\u00e9d\u00e9ric M.","family":"Vaz","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"},{"name":"Core Facility Metabolomics Amsterdam UMC Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mia L.","family":"Pras\u2010Raves","sequence":"additional","affiliation":[{"name":"Core Facility Metabolomics Amsterdam UMC Amsterdam North\u2010Holland Netherlands"},{"name":"University Medical Center Utrecht Utrecht Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karen Ghauharali\u2010van der","family":"Vlugt","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Femke S.","family":"Beers\u2010Stet","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susanna M. I.","family":"Goorden","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"},{"name":"Erasmus University Medical Center Rotterdam South\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Judith","family":"Jansen\u2010Meijer","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Georges E.","family":"Janssens","sequence":"additional","affiliation":[{"name":"Laboratory Genetic Metabolic Diseases Amsterdam UMC Location University of Amsterdam Amsterdam North\u2010Holland Netherlands"},{"name":"Amsterdam Gastroenterology Endocrinology and Metabolism Institute Amsterdam North\u2010Holland Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0464-1078","authenticated-orcid":false,"given":"Carla E. 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