Comparative proteomic analysis of human CD34⁺ stem/progenitor cells and mature CD15⁺ myeloid cells

Human CD34⁺ cells, highly enriched for hematopoietic stem and progenitors, and CD15⁺ cells, more terminally differentiated myeloid cells in blood, represent distinct maturation/differentiation stages. A proteomic approach was used to identify proteins differentially present in these two populations from human cord blood. Cytosolic proteins were extracted and subjected to two-dimensional gel electrophoresis followed by mass spectrometry. On average, 460 protein spots on each gel were detected; 112 and 15 proteins, respectively, were found to be differentially expressed or post-translationally modified in CD34⁺ and CD15⁺ cells. This suggests that CD34⁺ cells have a relatively larger proteome than mature CD15 ⁺ myeloid cells and production of many stem/progenitor cell-associated proteins ceases or is dramatically down-regulated as the CD34⁺ cells undergo differentiation. Of approximately 140 protein spots, 47 different proteins were positively identified by mass spectrometry and database search; these proteins belong to several functional categories, including cell signaling, transcription factors, cytoskeletal proteins, metabolism, protein folding, and vesicle trafficking. Multiple heat shock proteins and chaperones, as well as proteins important for intracellular trafficking, were predominantly present in CD34⁺ cells. Most of the identified proteins in CD34⁺ cells are expressed in germ cell tumors, as well as in embryonal carcinoma and neuroblastoma. Approximately eight novel proteins, whose functions are unknown, were identified. This study presents, for the first time, global cellular protein expression patterns in human CD34 ⁺ and CD15⁺ cells, which should help to better understand intracellular processes involved in myeloid differentiation and add insight into the functional capabilities of these distinct cell types.