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

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 prote...

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Bibliographic Details
Published in:International journal of molecular sciences 2018-01, Vol.19 (1), p.296
Main Authors: Wangen, Rebecca, Aasebø, Elise, Trentani, Andrea, Døskeland, Stein-Ove, Bruserud, Øystein, Selheim, Frode, Hernandez-Valladares, Maria
Format: Article
Language:English
Subjects:
Acute myeloid leukemia
Animals
Blood
Blood Proteins - chemistry
Buffers
Cattle
Communication
Contaminants
Cryoprotectors
Dimethyl sulfoxide
Freezing
Humans
Leukemia
Leukemia, Myeloid, Acute - genetics
Leukemia, Myeloid, Acute - pathology
Liquid nitrogen
Lysates
Mass Spectrometry
Mass spectroscopy
Myeloid leukemia
Peripheral blood
phosphate buffered saline
Phosphates - chemistry
Preservation
Proteins
Proteome - drug effects
Proteome - genetics
Proteomes
proteomics
Proteomics - methods
sample preparation
Sodium Chloride - pharmacology
Sodium dodecyl sulfate
Thawing
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Summary: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.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms19010296