ArticleMolecular therapy. Methods & clinical development2024
Extracellular vesicle depletion and UGCG overexpression mitigate the cell density effect in HEK293 cell culture transfection.
Article in Molecular therapy. Methods & clinical development, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Next-generation CAR-NK cell therapy: engineering strategies, translational challenges, and future perspectives in cancer immunotherapy.Cancer cell international · 2026Review
- Spent Medium Inhibits rVSV Infection.Viruses · 2026Article
- Development of a scalable production bioprocess for HIV-1 virus-like particles coupling continuous VLP harvesting with end-to-end downstream processing.Scientific reports · 2026Article
- Impact of specific productivity and operation mode upon the biophysical properties of HIV-1 Gag-based virus-like particles.Applied microbiology and biotechnology · 2025Article
- Perfusion-Based Production of rAAV via an Intensified Transient Transfection Process.Biotechnology and bioengineering · 2025Article
- Intensification of rAAV Production Based on HEK293 Cell Transient Transfection.Biotechnology journal · 2025Article
- Gag HIV-1 Virus-like Particles and Extracellular Vesicles Functionalization with Spike Epitopes of SARS-CoV-2 Using a Copper-Free Click Chemistry Approach.Bioconjugate chemistry · 2025Article
- Targeted UDP-glucose ceramide glucosyltransferase stable overexpression induces a metabolic switch improving cell performance at high cell density.Frontiers in bioengineering and biotechnology · 2025Article
- Unlocking DOE potential by selecting the most appropriate design for rAAV optimization.Molecular therapy. Methods & clinical development · 2024Article
- Harnessing cell aggregates for enhanced adeno-associated virus manufacturing: Cultivation strategies and scale-up considerations.Biotechnology progressArticle
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Authors and funding
6 authors.
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Abstract
The hitherto unexplained reduction of cell-specific productivity in transient gene expression (TGE) at high cell density (HCD) is known as the cell density effect (CDE). It currently represents a major challenge in TGE-based bioprocess intensification. This phenomenon has been largely reported, but the molecular principles governing it are still unclear. The CDE is currently understood to be caused by the combination of an unknown inhibitory compound in the extracellular medium and an uncharacterized cellular change at HCD. This study investigates the role of extracellular vesicles (EVs) as extracellular inhibitors for transfection through the production of HIV-1 Gag virus-like particles (VLPs) via transient transfection in HEK293 cells. EV depletion from the extracellular medium restored transfection efficiency in conditions that suffer from the CDE, also enhancing VLP budding and improving production by 60%. Moreover, an alteration in endosomal formation was observed at HCD, sequestering polyplexes and preventing transfection. Overexpression of UDP-glucose ceramide glucosyltransferase (UGCG) enzyme removed intracellular polyplex sequestration, improving transfection efficiency. Combining EV depletion and UGCG overexpression improved transfection efficiency by ∼45% at 12 × 10
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