title: Microenvironment-driven changes in the expression profile of hematopoietic cobblestone area-forming cells.
abstract: Studies with ex vivo cultures of bone marrow have indicated the importance of the adherent layer as a primary reservoir of the most primitive hematopoietic stem cells, from which derivative stem cells and more differentiated progenitors are continuously generated. We used the Affymetrix GeneChip to analyze the mRNA expressions between bone marrow-derived hematopoietic progenitor cells in the cobblestone areas (CA) and the free-floating cells released from the CA formations. Mouse bone marrow hematopoietic progenitor cell line FDCP-Mix and S17 stromal cells were used in this study. Of the 12,000 genes on the chip, only 29 showed more than fivefold higher in CAFC; and for cells in the supernatant, only 55 showed fivefold higher expressions than in the cobblestone area-forming cells (CAFC). The hematopoietic cells in CAFC expressed genes associated with homing, adhesion, and suppression of differentiation, while the free-floating hematopoietic cells showed mature lineage markers and differentiation-specific genes. This confirmed the more primitive nature of the hematopoietic cells in the adherent layer. Of interest in the findings were the discoveries of many secreted and surface protein expressions in CA hematopoietic cells. This may imply interactions among the hematopoietic cells, stromal cells, and the extracellular matrix in CA, which drive the growth, maturation, and differentiation of the hematopoietic cells.
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, #06-01, Centros, Singapore, Republic of Singapore. choong_meng_ling@bti.a-star.edu.sg
authors.forename: M L
authors.initials: ML
authors.lastname: Choong
authors.forename: B
authors.initials: B
authors.lastname: Luo
authors.forename: H F
authors.initials: HF
authors.lastname: Lodish
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D017154:Stromal Cells / Q000166:cytology