Abstract
For more than a decade numerous laboratories have differentiated osteoprogenitors from neonatal calvaria or marrow stromal cells/ mesenchymal stem cells (MSCs) from the bone marrow into mature osteoblasts in tissue culture. During this process, a single-cell or colony-forming unit (CFU) is able to proliferate, secrete, and mineralize bone matrix yielding a discrete raised nodule that protrudes into the medium and readily absorbs the silver nitrate in a von Kossa stain after fixation. Observing the similarities between the differentiation potential of rodent and human MSCs and murine embryonic stem (ES) cells, the ability of ES cells to form mineralized nodules in vitro under similar conditions is explored in the chapter. The author has utilized a fairly simple method for allowing EB formation and culture the EBs for only two days, after which individual cells are plated under conditions typically used for the in vitro differentiation of bone nodules from stem and progenitor cells from other sources. Despite the differences in the methodology, it is clear that there is a progenitor population in the EB that is capable of becoming an osteoprogenitor and forming a mineralized nodule in vitro. This is extremely important because ES cell-derived osteoblasts and their progenitors show excellent therapeutic promise for osteoporosis, rheumatic diseases, developmental bone defects, and severe fractures. Also, the in vitro differentiation of ES cells along the osteoblast lineage is a valuable tool for addressing pertinent questions about the proliferation, differentiation, survival, and intercellular communication between cells of the bone lineage.
Published Version
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