Production of Adult Human Synovial Fluid-Derived Mesenchymal Stem Cells in Stirred-Suspension Culture

Figure 8

Multipotent differentiation potential of SF-MSCs cultured for 18 days in spinner flasks using the bead-to-bead transfer method. Multipotency was evaluated for cells isolated from both donor 1 and donor 2. Shown here are photomicrographs for donor 1, which are representative of the qualitative results obtained for donor 2. (a) Osteogenic differentiation of donor 1 SF-MSCs. Calcium deposition is an indication of osteogenesis and was detected using Alizarin Red. Photomicrograph was taken at 10x magnification. Scale bar represents 200 μm. (b) Adipogenic differentiation of donor 1 SF-MSCs. Intracellular lipid droplet formation is an indication of adipogenesis and was detected using Oil Red O. Photomicrograph was taken at 20x magnification. Scale bar represents 100 μm. (c) Chondrogenic differentiation of donor 1. Glycosaminoglycan (GAG) production is an indication of chondrogenesis and results in an increase in pellet size (right pellet was induced; left pellet was not induced (control)). Photomicrograph was taken at 5x magnification. Scale bar represents 500 μm. (d) Quantitative chondrogenic differentiation analysis of donor 1 and donor 2 using the pellet culture method. 2.5 × 10⁵ cells/pellet were cultured in either 10% FBS DMEM (control) or chondrogenic induction medium (induced) for 28 days at 37°C and 5% CO₂. GAG production was quantified using 1,9-dimethylmethlylene blue after digesting the pellets in papain solution. Data were collected in triplicate; error bars represent the standard deviation of the data. compared to the control.

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