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Early proliferation of umbilical cord blood cells from premature neonates
Author(s) -
Luzo A. C. M.,
Duarte A. S. S.,
Salles T. S. I.,
Queiroz M. L. S.,
LorandMetze I.,
Costa F. F.,
Saad S. T. O.
Publication year - 2007
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1111/j.1423-0410.2007.00936.x
Subject(s) - andrology , progenitor cell , clonogenic assay , umbilical cord , haematopoiesis , flow cytometry , immunology , cord blood , biology , colony forming unit , cell cycle , stem cell , megakaryocyte , cell culture , microbiology and biotechnology , medicine , cell , biochemistry , genetics , bacteria
Background and Objective Human umbilical cord blood (UCB) is an important source of haematopoietic stem cells; however, the behaviour of progenitor cells obtained from premature and full‐term neonates is still a controversy subject. Thus, the aim of this study was to evaluate cell cycle parameters and the proliferative capacity of UCB progenitor cells from premature and full‐term neonates. Material and Methods Clonogenic assays were performed with methylcellulose, medium supplemented with recombinant stimulating growth factors and the colonies were scored on the seventh day and the 14th day of culture. A cell cycle study was carried out by DNA analysis using flow cytometry and 30 000 events were acquired; p107 and p130 expressions were analysed by Western blotting. Results Cultures obtained from UCB of premature neonates showed an early growth of colony‐forming unit (CFU)‐burst forming unit erythroid/CFU‐granulocyte, erythrocyte, macrophage and megakaryocyte (BFU‐E/GEMM), and CFU‐granulocyte, macrophage (GM) by the seventh day of culture ( P < 0·001). Therefore, the number and morphological characteristics of these colonies were comparable with those obtained from full‐term neonates, on the 14th day of culture. At the 14th day, a large amount of CFU‐GM was detected in the premature group ( P < 0·0032). The premature culture on the 14th day showed fibroblasts and was comparable to those of full‐term neonates on the 21st day in terms of number and morphology of the colonies. DNA analysis showed that the number of cells in S‐phase was also higher in premature samples when compared to full‐term neonates, P < 0·0021 (0 h = 12·8 vs. 2·5%; 16 h = 10·5 vs. 5·9%; 20 h = 13·5 vs. 10·3%; 24 h = 13·8 vs. 9·1%; 48 h = 14·0 vs. 5·4%; 72 h = 20·5 vs. 8·9%; 96 h = 13·8 vs. 7·7%). The Western blotting results demonstrated that p107 and p130 cell cycle protein expressions were higher in premature cells than in full‐term cells. Conclusion These results suggest that the higher capacity of proliferation and early differentiation of premature UCB might not be related only to the amount of stem/progenitor cells but also to a different timing of cell cycle entry.