{"doi":"10.1098/rsif.2023.0468","title":"A tumour-spheroid manufacturing and cryopreservation process that yields a highly reproducible product ready for direct use in drug screening assays","abstract":"If it were possible to purchase tumour-spheroids as a standardised product, ready for direct use in assays, this may contribute to greater research reproducibility, potentially reducing costs and accelerating outcomes. Herein, we describe a workflow where uniformly sized cancer tumour-spheroids are mass-produced using microwell culture, cryopreserved with high viability, and then cultured in neutral buoyancy media for drug testing. C4-2B prostate cancer or MCF-7 breast cancer cells amalgamated into uniform tumour-spheroids after 48 h of culture. Tumour-spheroids formed from 100 cells each tolerated the cryopreservation process marginally better than tumour-spheroids formed from 200 or 400 cells. Post-thaw, tumour-spheroid metabolic activity was significantly reduced, suggesting mitochondrial damage. Metabolic function was rescued by thawing the tumour-spheroids into medium supplemented with 10 µM N -Acetyl- l -cysteine (NAC). Following thaw, the neutral buoyancy media, Happy Cell ASM, was used to maintain tumour-spheroids as discrete tissues during drug testing. Fresh and cryopreserved C4-2B or MCF-7 tumour-spheroids responded similarly to titrations of Docetaxel. This protocol will contribute to a future where tumour-spheroids may be available for purchase as reliable and reproducible products, allowing laboratories to efficiently replicate and build on published research, in many cases, making tumour-spheroids simply another cell culture reagent.","journal":"Journal of The Royal Society Interface","year":2023,"id":375728,"datarank":0.2845561064841889,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"self_citation_contribution":0.24141568686511508,"citation_network_contribution":0.04314041961907382,"self_endowment_contribution":0.24141568686511508,"citer_contribution":0.04314041961907382,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":3,"citers_with_citation_signal":2,"citers_with_endowment":2,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.951,"is_data_producer":true,"deposit_databanks":{"figshare":["10.6084/m9.figshare. c.6845644"]},"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":574568,"name":"Kathryn Futrega","orcid":"0000-0003-3411-6768","position":1,"is_corresponding":false},{"id":1138634,"name":"Anthony M. Davies","orcid":null,"position":2,"is_corresponding":false},{"id":869837,"name":"Rose Ann Franco","orcid":"0000-0001-9772-714X","position":3,"is_corresponding":false},{"id":869838,"name":"Eamonn McKenna","orcid":"0000-0002-2324-592X","position":4,"is_corresponding":false},{"id":1097729,"name":"Bianca Guillesser","orcid":"0000-0002-4562-0401","position":5,"is_corresponding":false},{"id":399140,"name":"Travis J. Klein","orcid":"0000-0002-6669-7766","position":6,"is_corresponding":false},{"id":574570,"name":"Ross Crawford","orcid":"0000-0001-6079-1316","position":7,"is_corresponding":false},{"id":5753,"name":"Michael R. Doran","orcid":"0000-0001-5876-4757","position":8,"is_corresponding":false},{"id":869839,"name":"Md. Shafiullah Shajib","orcid":"0000-0002-1071-6868","position":0,"is_corresponding":true}],"reference_count":50,"raw_metadata":null,"created_at":"2026-07-19T01:16:23.381203Z","pmid":"37817581","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":[]}