{"doi":"10.3791/64271","title":"Co-Culture and Transduction of Murine Thymocytes on Delta-Like 4-Expressing Stromal Cells to Study Oncogenes in T-Cell Leukemia","abstract":"Transduced mouse immature thymocytes can be differentiated into T cells in vitro using the delta-like 4-expressing bone marrow stromal cell line co-culture system (OP9-DL4). As retroviral transduction requires dividing cells for transgene integration, OP9-DL4 provides a suitable in vitro environment for cultivating hematopoietic progenitor cells. This is particularly advantageous when studying the effects of the expression of a specific gene during normal T cell development and leukemogenesis, as it allows researchers to circumvent the time-consuming process of generating transgenic mice. To achieve successful outcomes, a series of coordinated steps involving the simultaneous manipulation of different types of cells must be carefully performed. Although these are very well-established procedures, the lack of a common source in the literature often means a series of optimizations are required, which can be time-consuming. This protocol has been shown to be efficient in transducing primary thymocytes followed by differentiation on OP9-DL4 cells. Detailed here is a protocol that can serve as a quick and optimized guide for the co-culture of retrovirally transduced thymocytes on OP9-DL4 stromal cells.","journal":"Journal of Visualized Experiments","year":2023,"id":408401,"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":0.9488,"is_data_producer":false,"deposit_databanks":null,"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":257719,"name":"Wenqing Li","orcid":"0000-0002-8336-5552","position":1,"is_corresponding":false},{"id":771776,"name":"Sarah D. Cramer","orcid":"0000-0003-4687-1971","position":2,"is_corresponding":false},{"id":771777,"name":"Hila Winer","orcid":"0000-0002-1064-0154","position":3,"is_corresponding":false},{"id":975041,"name":"Tu Chun Hsu","orcid":"0000-0001-7591-530X","position":4,"is_corresponding":false},{"id":1121234,"name":"Timothy Gower","orcid":null,"position":5,"is_corresponding":false},{"id":771778,"name":"Julie A. Hixon","orcid":"0000-0002-6505-2392","position":6,"is_corresponding":false},{"id":272369,"name":"Scott K. Durum","orcid":"0000-0003-1307-9582","position":7,"is_corresponding":false},{"id":509940,"name":"Gisele Rodrigues","orcid":"0000-0002-8526-2258","position":0,"is_corresponding":true}],"reference_count":26,"raw_metadata":null,"created_at":"2026-07-19T01:21:18.414003Z","pmid":"37358271","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":[]}