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An optimizing lentiviral titer determination assay based on Raji cells
Quan Chen1,2, Luoquan Ao1, Qing Zhao1
1State Key Laboratory of Trauma and Chemical Poisoning, Department of Stem Cell and Regenerative Medicine, Daping Hospital, Army Medical University Chongqing 400042, China.
American Journal of Translational Research
|June 19, 2025
Summary
Raji cells provide a more reliable method for lentiviral titration than traditional HEK293T cells, improving gene therapy vector production. This suspension cell approach enhances consistency and scalability for lentiviral vector quantification.
Area of Science:
- Gene Therapy
- Virology
- Cell Biology
Background:
- Accurate lentiviral titration is essential for optimizing gene therapy transduction efficiency.
- Conventional methods using human embryonic kidney 293T (HEK293T) cells have limitations in fidelity and reproducibility.
Purpose of the Study:
- To evaluate suspension cell lines, specifically Raji cells, as a superior platform for lentiviral titration.
- To compare the reliability and efficiency of Raji cells against HEK293T cells for lentiviral quantification.
Main Methods:
- Compared transduction efficiencies of HEK293T and Raji cells under various viral doses and infection conditions.
- Quantified lentiviral titers using quantitative polymerase chain reaction (qPCR) targeting specific human reference and viral genes (ALB, PCBP2, WPRE, gag).
Main Results:
- Raji cells exhibited significantly higher transduction efficiency than HEK293T cells, especially at low viral inputs and high cell densities.
- qPCR titration indicated substantially higher functional titers in Raji cells compared to HEK293T cells.
- Primary T cell infections using Raji-derived titers demonstrated enhanced sensitivity and saturation at lower viral loads.
Conclusions:
- Raji cells present a more reliable and efficient alternative to HEK293T cells for lentiviral titration.
- This suspension cell-based method can improve the scalability and consistency of lentiviral vector production for gene therapy applications.
