Lysosomal Acidification in Cultured Astrocytes Using Nanoparticles.

Methods Mol Biol

Department of Public Health & Caring Sciences/Molecular Geriatrics, Rudbeck Laboratory, Uppsala University, 751 85, Uppsala, Sweden.

Published: February 2018

AI Article Synopsis

  • Nanoparticles, when taken up by cells, localize in lysosomes, which can be manipulated to alter their environment.
  • Acidic nanoparticles are used to reduce the pH levels in lysosomes of cultured astrocytes, enhancing their ability to degrade materials.
  • To conduct these experiments, neural stem cells from embryonic mouse cortex are isolated and differentiated into astrocytes, ensuring the culture lacks professional phagocytes, and the results are analyzed using specific antibodies and LysoTracker dye.

Article Abstract

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.

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Source
http://dx.doi.org/10.1007/978-1-4939-6934-0_10DOI Listing

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