Metabolite Extraction from RPE Cells and Retinas Related to Retinitis Pigmentosa

Methods Mol Biol. 2023:2560:257-265. doi: 10.1007/978-1-0716-2651-1_24.

Abstract

The application of metabolomics in ophthalmology helps to identify new biomarkers and elucidate disease mechanisms in different eye diseases, as well as aiding in the development of potential treatment options. Extracting metabolites successfully is essential for potential further analysis using mass spectrometry. In this chapter, we describe how to extract metabolites from a variety of sources including (1) cells on a dish, (2) cell culture medium, and (3) tissues in vivo with and without stable isotope tracers. Samples prepared using this protocol are suitable for a range of downstream mass spectrometry analyses and are stable in solvent for weeks at -80 °C.

Keywords: Gas or liquid chromatography with mass spectrometric detection (GC-MS and LC-MS); Metabolites extraction; Metabolomics; RPE cells; Retina dissection.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Humans
  • Retinitis Pigmentosa*