Baud, A;
Little, D;
Wen, TQ;
Heywood, WE;
Gissen, P;
Mills, K;
(2019)
An Optimized Method for the Proteomic Analysis of Low Volumes of Cell Culture Media and the Secretome: The Application and the Demonstration of Altered Protein Expression in iPSC-Derived Neuronal Cell Lines from Parkinson’s Disease Patients.
Journal of Proteome Research
, 18
(3)
pp. 1198-1207.
10.1021/acs.jproteome.8b00831.
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Abstract
Traditionally, cell culture medium in iPSC-derived cell work is not the main focus of research and often considered as just "food for cells". We demonstrate that by manipulation of the media and optimised methodology, it is possible to use this solution to study the proteins that the cell secretes ("secretome"). This is particularly useful in the study of iPSC-derived neurons, which require long culture time. We demonstrate that media can be considered as the cellular equivalent of cerebrospinal fluid and can be used to model diseases with optimised incubation and sampling times. The ability not to sacrifice cells allows significant cost and research benefits. In this manuscript we describe an optimised method for the analysis of the cell media from iPSC-derived neuronal lines from control and Parkinson's disease patients. We have evaluated the use of standard and supplement B27-free cell media as well as five different sample preparation techniques for proteomic analysis of the cell secretome. Mass spectral analysis of culture media allowed for the identification of > 500 proteins, in 500 µL of media which is 20-40 less volume than reported previously. Using shorter incubation times and our optimized methodology, we describe the use of this technique to study and describe potential disease mechanisms in Parkinson's disease.
| Type: | Article |
|---|---|
| Title: | An Optimized Method for the Proteomic Analysis of Low Volumes of Cell Culture Media and the Secretome: The Application and the Demonstration of Altered Protein Expression in iPSC-Derived Neuronal Cell Lines from Parkinson’s Disease Patients |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1021/acs.jproteome.8b00831 |
| Publisher version: | https://doi.org/10.1021/acs.jproteome.8b00831 |
| Language: | English |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
| Keywords: | induced pluripotent stem cells; mass spectrometry; Parkinson’s disease; proteomics; secretome |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Genetics and Genomic Medicine Dept |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10068434 |
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