Identification of key genes responsible for cytokine-induced erythroid and myeloid differentiation and switching of hematopoietic stem cells by RAGE

Abstract

We utilized a unique culture system to analyze the expression patterns of gene, protein, and cell surface antigen, and the biological process of the related genes in erythroid and myeloid differentiation and switching of hematopoietic stem cells (HSCs) in response to cytokine alterations. Gene-specific fragments (266) identified from five populations of cytokine-stimulated HSCs were categorized into three groups: (1) expressed specifically in a single cell population; (2) expressed in two cell populations, and (3) expressed in three or more populations. Of 145 defined cDNAs, three (2%) were novel genes. Protein two-dimensional gel electrophoresis and flow cytometry analyses showed overlapped and distinguished protein expression profiles in the cell populations studied. Biological process mapping of mRNAs expressed in erythroid and myeloid lineages indicated that mRNAs shared by both lineages attended 'core processes,' whereas genes specifically expressed in either lineage alone were related to specific processes or cellular maturation. Data from this study support the hypothesis that committed HSCs (E14 or G14) cells can still be redirected to develop into myeloid or erythroid cells when erythropoietin (EPO) is replaced with granulocyte-colony stimulating factor (G-CSF) under erythroid-cultured condition or G-CSF with EPO in myeloid-cultured environment, respectively. Our results suggest that genes or proteins co-expressed in erythroid and myeloid lineages may be essential for the lineage maintenance and switching in hematopoiesis.Cell Research (2006) 16:923-939.

Identification of CD13+CD36+ cells as a common progenitor for erythroid and myeloid lineages in human bone marrow

“…Meanwhile, in both E14 and G14 cultures, CD133 + CD13 − CD36 − or CD34 + CD13 − CD36 − cells decreased to less than 1% from their original >97% at D0, which is consistent with a previous report [28]. It is unlikely that the minimal number of CD133 + CD13 − CD36 − or CD34 + CD13 − CD36 − cells at D14 can give rise to abundant erythroid or myeloid lineage cells at this point [14]. However, some of the CD13 + CD36 + cells may still express a low intensity of known stem cell markers (such as CD133 and CD34) or other unexpected marker(s).…”

“…In contrast, G-CSF had no significant effect on erythroid progenitors in G14, consistent with previous data [45]. The differential effects of EPO and G-CSF on erythroid and myeloid development may in part explain the observations in switch-culture experiments done previously [13][14][15], in which E14 cells contain a number of CD13 + cells, rather than being exclusively committed to erythroid lineage, while G14 cells treated with EPO for another 14 days instead of G-CSF still retained some myeloid progenitor cells.…”

“…All cells were grown in media that included stem cell factor (SCF, 50 ng/mL), interleukin-3 (IL-3, 10 ng/mL), granulocyte-macrophage colonystimulating factor (GM-CSF; 10 ng/mL), and 30% fetal bovine serum (HyClone, MN, USA) as described previously [14]. All growth factors are purchased from Stem Cell Technologies (Vancouver, BC, Canada).…”

“…To identify population(s) that contain common precursors for both myeloid and erythroid development, these cells were sorted based on their expression of CD13 and CD36 using the FACS Vantage system (Becton Dickinson). Sorted cells were assessed morphologically by Wright-Giemsa staining as described previously [14].…”

“…We have previously shown that when CD133 (also known as AC133)-selected hematopoietic stem/progenitor cells are induced to follow the erythroid developmental pathway using erythropoietin (EPO) treatment, the resulting cells can still be redirected to develop into myeloid cells when EPO is replaced by granulocyte-colony stimulating factor (G-CSF), or vice versa [13][14][15]. These results indicate that some bi-potential precursors may exist in EPOinduced erythroid or G-CSF-induced myeloid populations [16,17].…”