An Optical and Chemiluminescence Assay for Assessing the Cytotoxicity of Balamuthia mandrillaris against Human Neurospheroids.
Worakamol PengsartKasem KulkeawPublished in: Bioengineering (Basel, Switzerland) (2022)
A spheroid is a cell aggregate in a three-dimensional context; thereby, it recapitulates the cellular architecture in human tissue. However, the utility of spheroids as an assay for host-parasite interactions remains unexplored. This study demonstrates the potential use of neurospheroids for assessing the cytotoxicity of the life-threatening pathogenic amoeba Balamuthia mandrillaris . The neuroblastoma SH-SY5Y cells formed a spheroid in a hanging drop of culture medium. Cellular damage caused by B. mandrillaris trophozoites on human neuronal spheroids was observed using microscopic imaging and ATP detection. B. mandrillaris trophozoites rapidly caused a decrease in ATP production in the spheroid, leading to loss of neurospheroid integrity. Moreover, 3D confocal microscopy imaging revealed interactions between the trophozoites and SH-SY5Y neuronal cells in the outer layer of the neurospheroid. In conclusion, the neurospheroid allows the assessment of host cell damage in a simple and quantitative manner.
Keyphrases
- endothelial cells
- high resolution
- induced apoptosis
- single cell
- induced pluripotent stem cells
- oxidative stress
- cell cycle arrest
- pluripotent stem cells
- high throughput
- cell therapy
- stem cells
- signaling pathway
- endoplasmic reticulum stress
- climate change
- sensitive detection
- photodynamic therapy
- blood brain barrier
- plasmodium falciparum