[Establishment of two-dimensional gel electrophoresis profiles of proteome from CD34(+) hematopoietic stem/progenitor cells].
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.
Wei
Yong-qiang
YQ
Department of Hematology, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China. wyq@fimmu.com
Feng
Ru
R
Yi
Zeng-shan
ZS
Liu
Qi-fa
QF
D018952:Antigens, CD34 / Q000032:analysis*; D002469:Cell Separation; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006412:Hematopoietic Stem Cells / Q000737:chemistry* / Q000166:cytology; D006801:Humans; D008297:Male; D020543:Proteome
D018952:Antigens, CD34; D020543:Proteome