{"doi":"10.3389/fbioe.2025.1632810","title":"Alternating 2D and 3D culture reduces cell size and extends the lifespan of placenta-derived mesenchymal stem cells","abstract":"Background: Mesenchymal stem cells (MSCs) hold great promise for treating a variety of human diseases; however, their clinical translation is hindered by challenges in large-scale expansion while preserving therapeutic potency and maintaining small cell size. Conventional 2D culture on rigid substrates induces MSC senescence and enlargement, compromising their function and biodistribution. Methods: We present an alternating 2D/3D culture strategy that combines adherent monolayer expansion with transient spheroid formation to mitigate these limitations. Placenta-derived MSCs were cultured under optimized spheroid conditions, with extracellular matrix supplementation and chemically defined media to enhance viability. To address scalability, we developed RGD-functionalized alginate hydrogel tubes (AlgTubes) that enable dynamic transitions between adherent and spheroid states for continuous culture. Results: Spheroid culture significantly reduced cell size and enhanced immunomodulatory function. The alternating 2D/3D protocol slowed MSC enlargement and senescence over multiple passages while preserving anti-inflammatory activity. Extracellular matrix supplementation and chemically defined media further improved cell viability. AlgTubes successfully supported the alternating culture strategy in a continuous and scalable format. Conclusions: The alternating 2D/3D culture system effectively overcomes limitations of conventional MSC expansion by mitigating enlargement, delaying senescence, and preserving both proliferative capacity and immunoregulatory potency. Combined with AlgTube technology, this work demonstrates a promising strategy for MSC manufacturing.","journal":"Frontiers in Bioengineering and Biotechnology","year":2025,"id":546291,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.96,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"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":998287,"name":"Li Han","orcid":"0000-0002-2780-0978","position":1,"is_corresponding":false},{"id":1438035,"name":"Yakun Yang","orcid":"0000-0002-4772-4263","position":2,"is_corresponding":false},{"id":60709,"name":"Xinran Wu","orcid":"0000-0002-4974-1913","position":3,"is_corresponding":false},{"id":259298,"name":"Aijun Wang","orcid":"0000-0002-2985-3627","position":4,"is_corresponding":false},{"id":555971,"name":"Liangqi Xie","orcid":"0000-0002-6546-9622","position":5,"is_corresponding":false},{"id":281636,"name":"Wuqiang Zhu","orcid":"0000-0001-5217-7789","position":6,"is_corresponding":false},{"id":1359628,"name":"Shue Wang","orcid":"0000-0001-7382-5378","position":7,"is_corresponding":false},{"id":998293,"name":"Yuguo Lei","orcid":"0000-0002-7682-6912","position":8,"is_corresponding":false},{"id":1438034,"name":"Ying Pan","orcid":"0000-0001-9935-6194","position":0,"is_corresponding":true}],"reference_count":102,"raw_metadata":null,"created_at":"2026-07-19T02:53:32.285899Z","pmid":"40918436","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":[]}