Autophagy describes the breakdown of unnecessary and dysfunctional components inside of cells. It takes place in specialized vesicles, called Lysosomes. Autophagy is involved in various critical cellular functions such as cellular homeostasis, energy production, cell differentiation, aging, and also in programmed cell death. Thus understanding the molecular basis of autophagy is of huge interest. However, all available methodologies so far are very laborious and expensive.
Waleska K. Martins and colleagues, University of São Paulo, Brazil, established a technique that allows calculating a parameter that is sensitive and specific to cell deathy by autophagy. For this purpose they combined three different, commonly used in vitro assays evaluating the survival rates of cells. Two of them evaluate the cytotoxicity based on the growth rate of cells (MTT reduction assay and CVS assay), while the third assay (NRU assay) quantifies the alteration of the Lysosomes. Combining the measured values allows the quantitation of cell death occurring by autophagy.
A high throughput application of this technique permits the analysis of different potentially cytotoxic compounds helping to reveal new insights into the molecular basis of autophagy and autophagic cell death.
- Rapid screening of potential autophagic inductor agents using mammalian cell lines
Waleska K. Martins, Divinomar Severino, Cleidiane Souza, Beatriz S. Stolf, Maurício S. Baptista,
Biotechnol. J. 2013.