Following cellular engulfment, nanoparticles end up in the lysosomes, making them an ideal tool for modifying the lysosomal environment. Here, we describe how acidic nanoparticles can be used to lower the pH of lysosomes in cultured, primary astrocytes and thereby increase their degradation capacity. To guarantee that the cell culture is completely devoid of professional phagocytes, we isolate, expand, and differentiate neural stem cells from embryonic mouse cortex to achieve astrocytes for these experiments. Immunostainings with LAMP2-specific antibodies can be performed to verify the lysosomal localization of the nanoparticles, and the effect on lysosomal acidification can easily be followed with LysoTracker dye.
Keywords: Acidification; Astrocyte; Degradation; LAMP2; LysoTracker; Nanoparticles; Primary cell culture; pHrodo.