61P - Optimized protocol for the accelerated production of dendritic cell-derived exosomes (DEXs): Achieving speed without compromising efficacy

Background

Dendritic cell-derived exosomes (DEXs) represent an advanced cancer immunotherapy strategy, particularly in patients with advanced stages. However, the standard 14-day production time poses logistical challenges that limit access to this therapy. This study aimed to reduce the production time to 10 days, without compromising the immunological efficacy or structural integrity of the exosomes, to make the therapy more accessible and cost-effective.

Methods

To optimize DEX production, adjustments were made to the maturation times and redundant steps in cell culture were eliminated. Two production protocols were compared: an optimized 10-day protocol and the standard 14-day protocol. Three key parameters were evaluated:

1. Cell viability: This was measured by flow cytometry and cell viability assays.

2. Immunological potency: Assessed through the release of key cytokines associated with TH1 (IL-2, TNF-α, IFN-γ), TH2 (IL-4, IL-10), and TH17 (IL-6, IL-17A) profiles, using ELISA assays and flow cytometry.

3. Structural integrity: Assessed by western blot for expression of key exosomal markers (CD80, CD83, HLA-DR) and assessment of exosome size and morphology using Nanosight.

Results

Cell viability: The optimized 10-day protocol maintained an average cell viability of 91.2%, compared to 94.1% for the 14-day protocol. The 3% difference was not significant. Immunological potency: Differences in cytokine levels between the two protocols were less than 5%. Structural integrity: Differences in the expression of exosomal markers CD80, CD83, and HLA-DR were less than 5%. The average size of exosomes was 110 nm for the 10-day protocol and 112 nm for the 14-day protocol, with a percentage difference of 1.79%.

Conclusions

Reducing DEX production time from 14 to 10 days improves efficiency and reduces costs by 25.3%, without compromising quality. This protocol ensures cell viability, immunological potency, and exosomal integrity, making it ideal for centers with limited resources. Optimization allows scaling immunotherapy, increasing global access, essential pillars for its implementation.

Legal entity responsible for the study

Orlando Global R&D LLC.

Funding

Biotech Foundation.

Disclosure

All authors have declared no conflicts of interest.