2026-10-06T16:04:20Z https://api.figshare.com/v2/oai

oai:figshare.com:article/34042302 2026-10-01T09:58:41Z category_44 portal_316 item_type_2 month_year_10_2026

Video 3_Xeno-free production of lentiviral particles: implementing hPL as FBS replacement in cell culture.mp4 Mariana Soares Guedes (8232747) Marnie Cole (21441278) Daniela Yildiz (11851994) Toxicology 3Rs (reduce, replace, refine) lentivirus serum-free virus production serum-free adaptation <p>Commonly used for delivering fluorescent or bioluminescent reporters to target genes, lentiviral vectors are an “easy-to-clone” vehicle to study up- and downregulation of gene expression induced by drug inhibitors, cytokine stimulants, chemicals, and environmental toxicants. To produce viral particles safely, several plasmid constructs are delivered to human embryonic kidney cells (HEK293-T), which use the cellular machinery to package and assemble non-replicative lentiviruses with high efficiency. However, the cell culture of HEK293-T cells still largely relies on the use of fetal bovine serum for cell survival and proliferation. At large scale, this cell culture practice consumes a great amount of animal-derived serum, driving the need for continuous farming of animals whose fetuses would be used to obtain serum, posing grave ethical and practical concerns for human-relevant research. In this study, we present a feasible and scalable method to adapt the culture medium formulation for HEK293-T cells and demonstrate how virus production responds to this cell culture adaptation. Using a commercially available platelet-based supplement derived from human blood byproducts, we maintain cell viability and proliferation rates, with up to 82% confluence, and can produce lentiviruses (approximately 10⁶ infectious units/mL with up to 20% infectivity), while sparing the killing of bovine fetuses for serum collection, in a cost-efficient and humanized manner. Envisioning adoption in the pharma industry and large-scale virus production facilities, this simple yet robust cell medium formulation presents itself as an ethical and practical avenue to advance scientific innovation and bring one more layer of human relevance to biomedical research.</p> 2026-10-01T09:58:41Z Dataset Media 10.3389/ftox.2026.1903400.s002 https://figshare.com/articles/media/Video_3_Xeno-free_production_of_lentiviral_particles_implementing_hPL_as_FBS_replacement_in_cell_culture_mp4/34042302 CC BY 4.0