Characterization of a culture-derived CD15+CD11b- promyelocytic population from CD34+ peripheral blood cells

Abstract

Selected CD34+ cells from mobilized apheresis products were cultured in serum‐free or serum‐containing media supplemented with granulocyte colony‐stimulating factor (G‐CSF), granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), interleukin‐3 (IL‐3), and stem cell factor (SCF; c‐kit ligand). We examined the emergence of a CD15+ CD11b‐ population, which appeared morphologically to be promyelocytes. This CD15+CD11b‐ population can be further expanded in culture into morphologically mature granulocytes. In an attempt to characterize this culture‐derived CD15+CD11b‐ promyelocytic population, single cells were clone sorted into wells of a Terasaki plate containing various growth factors. We compared the growth factor requirements and kinetics of this apheresis culture‐derived CD15+ CD11b‐ population to the CD15+CD11b‐ population from fresh bone marrow samples. Our studies indicate that the CD15+CD11b‐ promyelocytic population from bone marrow and blood are equivalent in their ability to proliferate and in their requirements for growth factors. The CD15+CD11b‐ population in vitro shows a high proliferative capacity when compared with the other CD15/CD11b populations (CD15‐CD11b‐, CD15+CD11b+, CD15‐CD11b+). Thus, we can manipulate CD34+ cells in vitro to proliferate and differentiate toward a mature neutrophil lineage. The CD15+ CDllb‐ promyelocytic population derived from this culture may represent the most effective cultured cell population for therapeutic reduction of neutropenia in vivo based on both its stage of differentiation and its proliferative potential. J. Leukoc. Biol. 62: 480–484; 1997.

CD15, CD24, and CD29 Define a Surface Biomarker Code for Neural Lineage Differentiation of Stem Cells

“…CD15 (Lewis-X antigen, SSEA-1), and also CD133 (Prominin-1), have previously been identified as putative markers of stem cell identity not only in the neural system [15,22,42,43] but also in hematopoietic [44,45] and other [36,[46][47][48] cell types, suggesting functional relevance for ''stemness'' properties in general, for example for adherence to stem cell niches [49]. While CD15 and also the structurally related FORSE-1 antigen are strongly expressed on neuroepithelial cells, they are both downregulated upon further differentiation, which suggests that delamination from the neuroepithelial tube/rosette-like structures parallels surface expression changes of these glycolipid markers [50].…”

“…Work with hES cells was approved by the Partners Embryonic Stem Cell Research Oversight Committee. Undifferentiated human ES cell lines H7 (WA-07, XX, passages and H9 (WA-09, XX, passages [35][36][37][38][39][40][41][42][43][44][45] were cultured under growth conditions and passaging techniques previously described [7]. In vitro analysis was done with both hES cell lines, the transplantation assay was done with H9.…”

Influence of hemoglobin vesicles, cellular‐type artificial oxygen carriers, on human umbilical cord blood hematopoietic progenitor cells in vitro

“…These combinations of cytokines have been shown to promote proliferation and differentiation of CD34 þ cells toward mature erythroid and myeloid lineage cells, respectively. 22,23 Various concentrations of HbVs were added to the medium containing the cells. After 10 days incubation at 378C in a humidified atmosphere containing 5% CO 2 , the total cell counts were determined.…”

Biocompatibility Study of Hemoglobin Vesicles, Cellular-Type Artificial Oxygen Carriers, with Human Umbilical Cord Hematopoietic Stem/Progenitor Cells Using an In Vitro Expansion System

“…10,11 In addition, liquid culture system in a certain combination of hematopoietic cytokines allows hematopoietic progenitor cells to proliferate to single lineage cells such as erythroid and myeloid cells. 12,13 Using these systems, we have recently investigated the influence of HbVs on in vitro hematopoiesis of human hematopoietic progenitor cells obtained from umbilical cord blood (CB), and have demonstrated that longer exposure of CB progenitor cells to HbVs negatively affected progenitor activity, whereas such an effect was minimal under shorter exposure to HbVs. 14 The expansion of HSCs in culture while maintaining their functions has been extensively carried out.…”