Bone-marrow-derived mesenchymal stem cells (MSCs) have the potential to differentiate into a number of phenotypes, including\r\nadipocytes. Adipogenic differentiation has traditionally been performed in monolayer culture, and, while the expression of a fatcell\r\nphenotype can be achieved, this culture method is labor and material intensive and results in only small numbers of fragile\r\nadherent cells, which are not very useful for further applications. Aggregate culture is a cell-culture technique in which cells are\r\ninduced to form three-dimensional aggregates; this method has previously been used successfully, among others, to induce and\r\nstudy chondrogenic differentiation of MSCs. We have previously published an adaptation of the chondrogenic aggregate culture\r\nmethod to a 96-well plate format. Based on the success of this method, we have used the same format for the preparation of threedimensional\r\nadipogenic cultures. ?e MSCs differentiate rapidly, the aggregates can be handled and processed for histologic and\r\nbiochemical assays with ease, and the format offers signi??cant savings in supplies and labor. As a differentiation assay, this method\r\ncan distinguish between degrees of senescence and appears suitable for testing medium or drug formulations in a high-volume,\r\nhigh-throughput fashion.
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