Osteogenic imprinting upstream of marrow stromal cell differentiation

Five spontaneously transformed cell lines were established from a population of murine bone marrow stromal cells (BMSCs) and the expression profiles of phenotype-characteristic genes, patterns of in vitro differentiation, and osteogenic capacity after in vivo transplantation were determined for each. All the clones expressed stable levels of cbfa1, the osteogenic "master" gene, whereas the levels of individual phenotypic mRNAs were variable within each, suggestive of both maturational and phenotypic plasticity in vitro. Varying levels of collagen type I and alkaline phosphatase (AP) were expressed in all the clonal lines. The clonal lines with proven in vivo osteogenic potential (3 out of 5) had a high proliferation rate and expressed bone sialoprotein (BSP), whereas the two nonosteogenic clones proliferated more slowly and never expressed BSP. Bone nodules were only observed in 2 out of 3 of the osteogenic lines, and only 1 out of three formed cartilage-like matrix in vitro. There was no evidence of chondrogenesis in the nonosteogenic lines. By contrast, LPL was expressed in two osteogenic and in two nonosteogenic lines. These results demonstrate the presence of multipotential and restricted progenitors in the murine stromal system. cbfa1, collagen type I, and AP expression were common to all, and therefore presumably early, basic traits of stromal cell lines that otherwise significantly differ with respect to growth and differentiation potential. This finding suggests that an osteogenic imprinting lies upstream of diversification, modulation, and restriction of stromal cell differentiation potential.