[The diversity of human hematopoietic stem/progenitor cells: IX. Cell-cycles kinetics related macromolecules DNA, RNA and protein contents in CD34+ hematopoietic cells of human bone marrow].
DNA, RNA and PRO comprise the bulk of macromolecules in cells, which have been proven to play an important role in regulating cell cycle transverse capacity, cell division, growth, and size. Simultaneous analysis of these moieties could provide more comprehensive and accurate information on cell cycle kinetics. In this study, DNA, RNA and PRO contents related to cell cycle kinetics in CD34+ hematopoietic cells of human bone marrow were measured to understanding the cell cycle kinetic features in CD34+ hematopoietic cells. For this reason, CIMS-100 immunomagnetic isolator, a novel isolation system, was used to enrich efficiently CD34+ hematopoietic cells from human bone marrow. The purity of enriched CD34+ hematopoietic cells determined by both FACS and APAAP staining is ranging from 90%-95%. Cellular DNA, RNA and PRO were stained with fluorochromes propidium iodide, pyronin Y and fluorescein isothiocyante respectively The fluorescence intensities reflecting the DNA, RNA and PRO content of individual cell were analyzed in FACS can by different excitation wavelengthes. DNA, RNA and PRO contents in CD34+ hematopoietic cells were far lower than these of bone marrow mononuclear cells, only being 34 +/- 3% (DNA), 48 +/- 21% (RNA) and 62 +/- 14% (PRO) of BMMNCs respectively. Collectively, these data combined with previous results from both ours and others indicated that CD34+ hematopoietic cells are indeed an unique cell population, not only in reconstitute of hematopoietic and immunological functions, but also in cell cycle kinetics. This is, to our knowledge, the first detailed report on the analysis of DNA, RNA and PRO contents related to cell cycle kinetics in CD34+ hematopoietic cells. And the results provide more direct evidence that the majority of CD34+ hematopoietic cells are in resting state.
Xi
Y Z
YZ
Department of Transplantation Immunology, Affiliated 307 Hospital for Academy of Medical Sciences, Beijing 100039.
Zhang
Z X
ZX
Zhang
S X
SX
Kong
F H
FH
Li
X S
XS
Tang
P H
PH
Wei
W
W
Jin
L
L
Chen
X G
XG
D018952:Antigens, CD34 / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology*; D002453:Cell Cycle; D004247:DNA / Q000378:metabolism*; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D012313:RNA / Q000378:metabolism*
D018952:Antigens, CD34; D001798:Blood Proteins; D012313:RNA; D004247:DNA