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Efficacy Evaluation of “Enhanced” Natural Killers with CISH and B2M Knockouts on Viability and Metabolic Status of 3D Glioblastoma Spheroid Cells in Patients

2025/02/28 by Diana V. Yuzhakova, D A Sachkova, Marina V. Shirmanova +5 · 1 voice
Biochemistry, Genetics and Molecular Biology · Immunology and Microbiology · Medicine · #Cancer Research and Treatments #Immune Cell Function and Interaction #Cancer Cells and Metastasis

paper · pdf · doi:10.17691/stm2025.17.1.10

openalex publication_date 2025/02/28 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/03

Abstract

was to evaluate the impact efficacy of "enhanced" NK cells on early metabolic rearrangements and the viability of glioblastoma cells in a patient using a tumor spheroid model. Materials and Methods: ) knockouts. Tumor spheroids were produced in round-bottomed, low-adhesive plates. 100 thousand immune cells were added to each spheroid, and spheroids viability was evaluated at several time points applying fluorescence staining using a live/dead cell viability assay kit; autofluorescence of metabolic coenzyme nicotinamide adenine dinucleotide (phosphate), or NAD(P)H, was visualized in spheroids using an LSM 880 laser scanning microscope (Carl Zeiss, Germany) with a FLIM module (Becker & Hickl GmbH, Germany). Results: , indicating occurrence of an early metabolic shift in tumor cells towards a less aggressive oxidative phenotype, and this is consistent with dead cells fraction increase and living cells fraction decrease in spheroid composition. Conclusion: The data obtained on enhanced cytotoxic activity of new modified NK cell lines against human glioblastoma spheroids are important to understand interaction mechanisms between tumor and immune cells and the development of glioblastoma adoptive cell therapy.

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