{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T05:17:16Z","timestamp":1788585436683,"version":"build-2803163510"},"reference-count":49,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Science Foundation of China","doi-asserted-by":"publisher","award":["32330083"],"award-info":[{"award-number":["32330083"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Science Foundation of China","doi-asserted-by":"publisher","award":["32500668"],"award-info":[{"award-number":["32500668"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010857","name":"Jiangxi Province National Science and Technology Prizes","doi-asserted-by":"publisher","award":["20212AEI91001"],"award-info":[{"award-number":["20212AEI91001"]}],"id":[{"id":"10.13039\/501100010857","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100022957","name":"Double Thousand Plan of Jiangxi Province","doi-asserted-by":"crossref","award":["jxsq2023102239"],"award-info":[{"award-number":["jxsq2023102239"]}],"id":[{"id":"10.13039\/100022957","id-type":"DOI","asserted-by":"crossref"}]},{"award":["jxsq2023102239"],"award-info":[{"award-number":["jxsq2023102239"]}],"id":[{"id":"https:\/\/ror.org\/03q269m48","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004479","name":"Natural Science Foundation of Jiangxi Province","doi-asserted-by":"publisher","award":["20242BAB25409"],"award-info":[{"award-number":["20242BAB25409"]}],"id":[{"id":"10.13039\/501100004479","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Background: Cultured primary mouse hepatocytes undergo drastic phenotypic and metabolic reprogramming, while the temporal rules and regulatory machinery of membrane phospholipid remodeling remain elusive. Methods: Relying on a 0\u201372 h time-series in vitro culture system, this study integrated multi-omics technologies to dissect the temporal dynamics of phospholipid remodeling in hepatocytes. Through phosphatidylethanolamine N-methyltransferase (PEMT) knockout, exogenous PEMT expression, and methionine deprivation, we examined the association of PEMT status and methionine availability with phospholipid remodeling. Results: In vitro cultivation reduces intracellular total phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE) through three coordinated events: suppressed transcription of phospholipid synthetic genes hinders de novo synthesis, elevated lipid hydrolysis consumes cellular phospholipids, and extracellular phospholipids accumulate in the culture medium from 12 to 48 h. These jointly trigger ordered remodeling of PC\/PE balance, acyl chain length and fatty acid unsaturation. PEMT knockout was associated with PE retention without worsening hepatocyte dedifferentiation, PEMT exogenous expression raises PC content and PC\/PE ratio yet cannot rescue culture-dominated lipid structural shifts. Methionine depletion depleted cellular methionine, S-adenosylmethionine (SAM) and S-adenosyl-L-homocysteine (SAH), producing selected lipid changes that partially overlapped with lipid phenotypes of PEMT knockout. Conclusion: In short, culture duration was the dominant factor associated with the fundamental phospholipid remodeling trajectory, and PEMT status and methionine availability, were associated with selective differences in lipid composition.<\/jats:p>","DOI":"10.3390\/cells15171608","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T16:21:21Z","timestamp":1788452481000},"page":"1608","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Time-Dependent Phospholipid Remodeling in Cultured Primary Mouse Hepatocytes: Associations with PEMT Status and Methionine Availability"],"prefix":"10.3390","volume":"15","author":[{"given":"Tianxin","family":"Ma","sequence":"first","affiliation":[{"name":"College of Chemistry and Materials, Jiangxi Normal University, Nanchang 330022, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kunpeng","family":"Zhou","sequence":"additional","affiliation":[{"name":"Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology, Life Sciences Institute, Zhejiang University, Hangzhou 310058, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Yibing","family":"Zhu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Reproductive Genetics (Ministry of Education) and Department of Reproductive Endocrinology, Women\u2019s Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"given":"Jiale","family":"Zhang","sequence":"additional","affiliation":[{"name":"Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology, Life Sciences Institute, Zhejiang University, Hangzhou 310058, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Xing","family":"Xie","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Natl R&D Branch Ctr Convent Freshwater Fish Proc, Jiangxi Normal University, Nanchang 330022, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"given":"Lu","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Natl R&D Branch Ctr Convent Freshwater Fish Proc, Jiangxi Normal University, Nanchang 330022, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3117-8680","authenticated-orcid":false,"given":"Cunqi","family":"Ye","sequence":"additional","affiliation":[{"name":"Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology, Life Sciences Institute, Zhejiang University, Hangzhou 310058, China"},{"name":"College of Life Sciences, Natl R&D Branch Ctr Convent Freshwater Fish Proc, Jiangxi Normal University, Nanchang 330022, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"given":"Zong-Cai","family":"Tu","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Natl R&D Branch Ctr Convent Freshwater Fish Proc, Jiangxi Normal University, Nanchang 330022, China"},{"name":"State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang 330047, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"114008","DOI":"10.1016\/j.yexcr.2024.114008","article-title":"A comparison between different human hepatocyte models reveals profound differences in net glucose production, lipid composition and metabolism in vitro","volume":"437","author":"Bonanini","year":"2024","journal-title":"Exp. 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