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

Figure 6

Bead-to-bead transfer of SF-MSCs on Cytodex 3 microcarriers. 1 g/L of fresh microcarriers was added to the culture on days 6 and 12 (shown with arrows). After addition, the cultured were stirred intermittently at 40 rpm for 3 hours (3 minutes on, 27 minutes off). After 3 hours, the culture was continuously stirred at 40 rpm. Shown are photomicrographs (a) immediately following addition of fresh microcarriers (day 6). Roughly half of the beads are empty and half are occupied and nearing confluence; (b) 3 hours after microcarrier addition on day 6. Cells had started to transfer to new microcarriers, but still appeared rounded on their surface; (c) 24 hours after microcarrier addition. More noticeable cell transfer had occurred and very few beads remained unoccupied; (d) 6 days following inoculation (day 12, prior to second addition of fresh microcarriers). Most beads were near confluence. Photomicrographs were taken at 10x magnification. Scale bars represent 200 μm. (e) Growth curve of bead-to-bead transfer of SF-MSCs on Cytodex 3 microcarriers. Shown are data for cells derived from donors 1 and 2 to show that the utility of the protocol developed was not unique for only a single set of cells. The increases in viable cell densities show that the bead-to-bead transfer method was successful. Data were collected in duplicate; error bars represent the range of data collected.

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