{"doi":"10.1002/cph4.70082","title":"Geltrex‐Enhanced Two‐Dimensional Culture as a Viable Alternative to Primary Rat Hepatocyte Sandwich Models","abstract":"Primary hepatocytes rapidly lose viability and function in conventional two-dimensional (2D) cultures due to the absence of a physiologically relevant extracellular matrix (ECM). The collagen sandwich method improves polarization and function but creates a diffusion barrier that limits nutrient and signal exchange. This study investigates whether daily supplementation of a diluted, non-gelling Geltrex layer can sustain hepatocyte function and viability in 2D culture, offering a practical alternative to the sandwich method. Primary rat hepatocytes were cultured for 15 days under four conditions: monolayer (ML), monolayer with Geltrex (ML + GT), sandwich (SW), and sandwich with Geltrex (SW + GT). Cell morphology, confluency, viability (CCK-8, live/dead staining), and functionality (urea synthesis, albumin production, CYP3A4 activity) were assessed. The ML group showed significant declines in confluency, viability, and functional markers over time. Geltrex supplementation preserved confluency (~97% at day 15), improved viability, and maintained higher albumin production and CYP3A4 activity compared to ML. Functional outputs in ML + GT were comparable to SW and SW + GT groups, without the diffusion limitations of the sandwich top gel. Daily supplementation with low-dose Geltrex creates a biochemically enriched, diffusion-permissive microenvironment that supports long-term viability and function of primary rat hepatocytes in 2D culture. This method represents a simple and effective alternative to traditional sandwich cultures for liver cell studies and drug testing applications.","journal":"Comprehensive physiology","year":2025,"id":584371,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9606,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1491388,"name":"Matilda Holtz","orcid":null,"position":1,"is_corresponding":false},{"id":1497207,"name":"Adi̇l Deni̇zli̇","orcid":"0000-0001-7548-5741","position":2,"is_corresponding":false},{"id":465503,"name":"O. Berk Usta","orcid":"0000-0001-9328-568X","position":3,"is_corresponding":false},{"id":1497206,"name":"Eda Beldek","orcid":"0009-0005-3653-6493","position":0,"is_corresponding":true}],"reference_count":69,"raw_metadata":null,"created_at":"2026-07-19T02:59:11.978098Z","pmid":"41351217","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}