Platelet-derived growth factor-BB (PDGF-BB) induces differentiation of bone marrow endothelial progenitor cell-derived cell line TR-BME2 into mural cells, and changes the phenotype

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Abstract
Blood vessels are composed of endothelial cells (EC) and mural cells, and the interaction between EC and mural cells is essential for the development and maintenance of the vasculature. EC differentiate from bone marrow‐derived endothelial progenitor cells (EPC). Recently, we established a conditionally immortalized bone marrow EPC‐derived cell line, TR‐BME2, and a brain capillary EC (BCEC) line, TR‐BBB, from temperature‐sensitive‐SV40 T‐antigen gene transgenic rats. To understand the function of EPC, it is important to analyze the difference between EPC and mature EC such as BCEC. In this study, we identified EPC‐specific genes by means of subtractive hybridization between TR‐BME2 and TR‐BBB. There was no significant difference between TR‐BME2 and TR‐BBB in the mRNA level of annexin II, which is expressed in EC. In contrast, the mRNA level of smooth muscle cell (SMC) markers such as smooth muscle protein 22 (SM22), calvasculin, and platelet‐derived growth factor (PDGF) receptor‐β, was higher in TR‐BME2 than in TR‐BBB. Moreover, the mRNA level of contractile SMC markers, such as smooth muscle α‐actin and SM22, was increased in the absence of EC growth factors, such as vascular endothelial growth factor. The mRNA level of synthetic SMC markers, such as matrix Gla protein, was increased by the addition of PDGF‐BB. The SMC derived from TR‐BME2 showed an altered phenotype, from contractile‐type to synthetic‐type, when they were cultured in the absence of PDGF‐BB. These results show that TR‐BME2 cells have higher levels of SMC markers compared with mature EC, and can differentiate into contractile‐ or synthetic‐type SMC.