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Int. J. Mol. Sci. 2018, 19(1), 296; doi:10.3390/ijms19010296

Preservation Method and Phosphate Buffered Saline Washing Affect the Acute Myeloid Leukemia Proteome

1
The Proteomics Unit at the University of Bergen, Department of Biomedicine, University of Bergen, Jonas Lies vei 91, 5020 Bergen, Norway
2
Department of Molecular Biology, University of Bergen, Thormøhlensgate 55, 5008 Bergen, Norway
3
Department of Clinical Science, University of Bergen, Jonas Lies vei 87, 5021 Bergen, Norway
4
Department of Biomedicine, University of Bergen, Jonas Lies vei 91, 5020 Bergen, Norway
*
Author to whom correspondence should be addressed.
Received: 12 December 2017 / Revised: 10 January 2018 / Accepted: 12 January 2018 / Published: 19 January 2018
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Abstract

Acute myeloid leukemia (AML) primary cells can be isolated from peripheral blood, suspended with media containing bovine serum and cryoprotectant, and stored in liquid nitrogen before being processed for proteomic analysis by mass spectrometry (MS). The presence of bovine serum and human blood proteins in AML samples can hamper the identifications of proteins, and thereby reduce the proteome coverage of the study. Herein, we have established the effect of phosphate buffered saline (PBS) washing on AML patient samples stored in media. Although PBS washes effectively removed serum and blood contaminants, the saline wash resulted in cell burst and remarkable protein material loss. We also compared different methods to preserve the AML proteome from THP-1 and Molm-13 cell lines before MS analysis: (1) stored in media containing bovine serum and dimethyl sulfoxide (DMSO); (2) stored as dried cell pellets; and (3) stored as cell lysates in 4% sodium dodecyl sulfate (SDS). MS analysis of differently preserved AML cell samples shows that preservation with DMSO produce a high number of fragile cells that will burst during freezing and thawing. Our studies encourage the use of alternative preservation methods for future MS analysis of the AML proteome. View Full-Text
Keywords: proteomics; acute myeloid leukemia; preservation; phosphate buffered saline; dimethyl sulfoxide; mass spectrometry; sample preparation proteomics; acute myeloid leukemia; preservation; phosphate buffered saline; dimethyl sulfoxide; mass spectrometry; sample preparation
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MDPI and ACS Style

Wangen, R.; Aasebø, E.; Trentani, A.; Døskeland, S.-O.; Bruserud, Ø.; Selheim, F.; Hernandez-Valladares, M. Preservation Method and Phosphate Buffered Saline Washing Affect the Acute Myeloid Leukemia Proteome. Int. J. Mol. Sci. 2018, 19, 296.

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