Committed differentiation of hematopoietic cells from human umbilical cord blood in vitro / 第三军医大学学报

ABSTRACT

Objective To explore the capacity of hematopoietic cells from human umbilical cord blood (UCB) induced by hematopoietic growth fators (HGFS) to differentiate committedly to granulocyte in vitro . Methods Suspension culture of isolated mononuclear cells from fresh UCB samples were conducted for 5 weeks in IMDM medium containing UCB serum and HGFS. The expansion folds of nucleated cells, the changes of CD34+antigen by immunofluorometry, and the colony forming unit granulocyte macrophage(CFU GM) by progenitor clonogenic assay were observed. Results Culture of mononuclear cells with the combination of granulocyte colony stimulating factor(G CSF), granulocyte macrophage colony stimulating factor (GM CSF), stem cell factor(SCF), interluekin 3(IL 3) and interleukin 6(IL 6) for 3 weeks markedly increased the total count of nucleated cells and CFU GM, being 58.3 and 12.2 folds of the pre cultured, and the cultured cells were mainly neutrophils. However, CD34+ cells in nucleated cells declined from 0.97% to 0.05%. According to the culture condition, a single portion of fresh UCB could increase nucleated cells by (5.1?2.3) fold and CFU GM by (2.2?0.95) fold. Conclusion Hematopoietic cells of UCB may be induced to differentiate committedely to granulocyte lineage by this culture system in vitro to result in the increase of granulocytes and CFU GM in single portion of cord blood.