Effects of Injectable Solutions on the Quality of Monocyte-Derived Dendritic Cells for Immunotherapy.
Laís Teodoro da SilvaBruna Tiaki TiyoSilvia de Jesus MotaMarina Mazzilli OrtegaGabriela J H SchmitzNoemi Nosomi TaniwakiGislene Mitsue NamiyamaAlberto Jose da Silva DuarteTelma Miyuki OshiroPublished in: Journal of immunology research (2024)
Therapeutic vaccines based on monocyte-derived dendritic cells have been shown to be promising strategies and may act as complementary treatments for viral infections, cancers, and, more recently, autoimmune diseases. Alpha-type-1-polarized dendritic cells (aDC1s) have been shown to induce type-1 immunity with a high capacity to produce interleukin-12p70 (IL-12p70). In the clinical use of cell-based therapeutics, injectable solutions can affect the morphology, immunophenotypic profile, and viability of cells before delivery and their survival after injection. In this sense, preparing a cell suspension that maintains the quality of aDC1s is essential to ensure effective immunotherapy. In the present study, monocytes were differentiated into aDC1s in the presence of IL-4 and GM-CSF. On day 5, the cells were matured by the addition of a cytokine cocktail consisting of IFN- α , IFN- γ , IL-1 β , TNF- α , and Poly I:C. After 48 hr, mature aDC1s were harvested and suspended in two different solutions: normal saline and Ringer's lactate. The maintenance of cells in suspension was evaluated after 4, 6, and 8 hr of storage. Cell viability, immunophenotyping, and apoptosis analyses were performed by flow cytometry. Cellular morphology was observed by electron microscopy, and the production of IL-12p70 by aDC1s was evaluated by ELISA. Compared with normal saline, Ringer's lactate solution was more effective at maintaining DC viability for up to 8 hr of incubation at 4 or 22°C.
Keyphrases
- dendritic cells
- cell cycle arrest
- induced apoptosis
- immune response
- regulatory t cells
- flow cytometry
- diffusion weighted
- diffusion weighted imaging
- cell death
- endoplasmic reticulum stress
- rheumatoid arthritis
- single cell
- sars cov
- stem cells
- signaling pathway
- pi k akt
- peripheral blood
- magnetic resonance
- electron microscopy
- contrast enhanced
- cell proliferation
- bone marrow
- ultrasound guided
- tissue engineering