Molecular study of hematopoiesis-supporting ability of C3H10T1/2 mouse embryo fibroblasts

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To investigate the molecular aspects of hematopoiesis-supporting ability of stromal cells, we used a differentiation-inducible mouse embryo fibroblast cell line, C3H10T1/2 (10T1/2). Stably determined preadipocyte and myoblast cell lines were established after a brief exposure of 10T1/2 cells to 5-azacytidine. These cell lines terminally differentiated into adipocytes and myotubes, respectively, under appropriate conditions. The hematopoiesis-supporting ability was significantly elevated at the preadipocyte stage (A54 preadipocyte), and was reduced after terminal adipocytic differentiation. To identify molecules that contribute to the hematopoiesis-supporting ability of preadipocytes, we screened genes that were differentially expressed in A54 preadipocytes and isolated the novel gene by mRNA differential display method. This gene was defined as a gene that was down-regulated during adipocyte differentiation-1 (drad-1). The drad-1 was expressed in other mouse preadipocytes, namely, ST2 and PA6 cells, that also have hematopoiesis-supporting ability. Moreover, the drad-1 was found to be expressed in mouse bone marrow. However, the function of the protein encoded by drad-1 is currently unknown. In addition, we investigated the change in CD34 mRNA expression during stromal cell differentiation. As a result, CD34 mRNA was constitutively expressed by parent 10T1/2 cells but not by adipogenically or myogenically determined cells. This finding supports the concept that CD34 may also be a marker of stromal progenitors and is lost as the cells differentiate into phenotypically distinct stromal elements. The 10T1/2-derived cell lines could provide a valuable tool to aid in the analysis of stromal cell development and differentiation and the search for novel stromal cell-derived factor (s).