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Differential gene expression in human hematopoietic stem cells specified toward erythroid, megakaryocytic, and granulocytic lineage

To better understand the transcriptional program that accompanies orderly lineage‐specific hematopoietic differentiation, we analyzed expression changes during the lineage‐specific differentiation of human hematopoietic stem cells (HSC; CD34+/CD38–/CD33–); HSC and multipotent myeloid progenitors (MMP; CD34+/CD38–/CD33+) were isolated from the bone marrow of healthy individuals by MACS. CD34+ cells in semi‐solid culture were stimulated with the cytokines erythropoietin, IL‐6, and G‐CSF to promote differentiation to committed erythroid, megakaryocytic, and granulocytic clones, respectively. Differential display RT‐PCR analysis was performed to compare the mRNA transcripts in HSC, MMP, and the committed lineage‐specific clones derived from these committed lineage‐specific progenitors. Expressed sequence tags (n=256), which were differentially expressed, were identified. One hundred ninety‐four were homologous to known genes, and some were associated with hematopoiesis. These known genes were classified as involved in transcription/translation, signal transduction, cell surface receptors/ligands, cell signaling, cell metabolism, cell cycle, cell apoptosis, and oncogenesis. We identified genes, which were up‐ or down‐regulated specifically in the lineage‐committed clones compared with HSC or/and MMP, suggesting that specific gene activation and repression might be necessary for specific lineage commitment and differentiation. Our data provide an extensive transcriptional profile of human hematopoiesis during in vitro, lineage‐specific differentiation.