Objective: To investigate the differential expression of proteins in bone marrow (BM) and mobilized peripheral blood (MPB) CD34+ cells.
Methods: Immunomagnetic beads were used to separate and purify CD34+ cells from the BM and the MPB mononuclear cells (MNCs) mobilized by granulocyte colony-stimulating factor (G-CSF) of normal subjects. The whole cell proteins in CD34+ cells were extracted by freeze-thaw lysis, and applied for two-dimensional electrophoresis (2-DE), the results analyzed with image analysis software.
Results: The average purity of CD34+ cells was 92.33%+/-2.65% in output, with an average cell number of (1.12+/-0.42) x 10(6). 2-DE techniques were optimized, which yielded satisfactory 2-DE profiles of the proteome, showing the different expressions of the proteins between different CD34+ cells.
Conclusions: Immunomagnetic beads in combination with 2-DE is applicable for research of hematopoietic stem/progenitor cells proteome. There are differences in the protein expressions between BM- and MPB-derived CD34+ cells.