Representational difference analysis of a committed myeloid progenitor cell line reveals evidence for bilineage potential

Abstract

In this study we have sought to characterize a committed myeloid progenitor cell line in an attempt to isolate general factors that may promote differentiation. We used cDNA representational difference analysis (RDA), which allows analysis of differential gene expression, to compare EML and EPRO cells. We have isolated nine differentially expressed cDNA fragments as confirmed by slot blot, Northern, and PCR analysis. Three of nine sequences appear to be novel whereas the identity of the remaining fragments suggested that the EPRO cell line is multipotent. Among the isolated sequences were eosinophilic, monocytic, and neutrophilic specific genes. Therefore, we tested the ability of EPRO cells to differentiate along multiple myeloid lineages and found that EPRO cells exhibited morphologic maturation into either monocyte͞macrophages or neutrophils, but not eosinophils. Furthermore, when EPRO cells were exposed to ATRA, neutrophil specific genes were induced, whereas monocytic markers were induced by phorbol ester treatment. This study highlights the use of cDNA RDA in conjunction with the EML͞EPRO cell line to isolate markers associated with macrophage and neutrophil differentiation and establishes the usefulness of this system in the search for factors involved in myeloid commitment.

Hoxa9 Immortalizes a Granulocyte-Macrophage Colony-Stimulating Factor-Dependent Promyelocyte Capable of Biphenotypic Differentiation to Neutrophils or Macrophages, Independent of Enforced Meis Expression

“…Similar to the majority of human AML (categories M1 to M5 [22,51]), the human promyelocytic HL60 cell line, and the murine 32Dcl3 cell line (45), they express MPO and NE but fail to express secondary granule proteins. Like HL60 and mouse EPRO cells, Hoxa9-immortalized promyelocytes retain bilineage differentiation potential (32) and, like EPRO cells treated with ATRA or 32Dcl3 cells grown in G-CSF, can be induced to undergo neutrophil differentiation accompanied by expression of a subset of secondary granule genes. Among cells with bilineage potential, however, Hoxa9-immortalized progenitors are unique in that they undergo cytokine-inducible morphologic differentiation to neutrophils (G-CSF) or macrophages (M-CSF) accompanied by the expression of selected secondary granule proteins or the F4/80 antigen and the macrophage acetyl-LDL scavenger receptor.…”

Insertional mutagenesis identifies genes that promote the immortalization of primary bone marrow progenitor cells

“…In response to GM-CSF, the cells continued to expand but significantly more macrophage (up to 30%) and neutrophil (up to 70%) differentiation occurred ( Figure 1C). The cells did not survive in the presence of M-CSF; however, treatment of the cells with PMA, a chemical that induces monocytic differentiation, 17 induced 50% to 70% of the cells to differentiate into macrophages ( Figure 1C). These cells thus demonstrate a partial differentiation block that can be overcome by growth factor addition.…”

Lactotransferrin-Cre reporter mice trace neutrophils, monocytes/macrophages and distinct subtypes of dendritic cells

“…34 Interestingly, EPRO cells -an Ltf-expressing promyelocytic cell line 37 -can be differentiated into either monocytes/macrophages or neutrophils, but not into eosinophils. 38 It remains to be determined whether these progenitors express Ltf in human hematopoiesis and -if so -whether Ltf + myeloid progenitors may give rise to neutrophils, monocytes, macrophages and subpopulations of DCs, while lacking the potential to develop into eosinophils, basophils or megakaryocytes.…”