Discovery proteomics by mass spectrometry comparing secreted proteins from young and old mouse lung mesenchymal stromal cells

Author ORCID

Diptiman Chanda 0000-0002-4835-4460

Publication Date

8-25-2021

Abstract

Lung mesenchymal stromal cells (L-MSCs) were isolated from the young ( 3 months) and old (22-24 months) mice (N = 4 each) following collagenase digestion of the lungs and anchorage-dependent growth. Culture media was collected from young and old L-MSCs (10^6 cells) grown in culture for 24 hrs in serum-free condition. Culture media was then centrifuged to remove any cellular debris. Five (5) ml of supernatants were concentrated using 3kDa Molecular weight cut-off filters, and 10 micrograms of proteins were run as multiple MW 1D PAGE fractions using a standard GeLC approach with a nano-HPLC in-line with a Velos Pro Orbitrap MS. Individual data files were searched using Sequest, the results were then combined for each sample, followed by grouping, filtering, and quantifying by normalized spectral counts.

Keywords

Secreted proteins, mesenchymal stromal cells, lung, mass spectrometry, proteomics

Repository

Dryad

Distribution License

CC0 1.0 Universal - No copyright; this work is in the public domain

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