IJMS, Vol. 19, Pages 296: Preservation Method and Phosphate Buffered Saline Washing Affect the Acute Myeloid Leukemia Proteome

IJMS, Vol. 19, Pages 296: Preservation Method and Phosphate Buffered Saline Washing Affect the Acute Myeloid Leukemia Proteome

International Journal of Molecular Sciences doi: 10.3390/ijms19010296

Authors: Rebecca Wangen Elise Aasebø Andrea Trentani Stein-Ove Døskeland Øystein Bruserud Frode Selheim Maria Hernandez-Valladares

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.

from #AlexandrosSfakianakis via Alexandros G.Sfakianakis on Inoreader http://ift.tt/2FRAXJw
via IFTTT