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. 2020 Dec 1;6:35. doi: 10.1038/s41526-020-00125-5

Fig. 3. Experimental design of combined SMG, LIVAT, and LIVDT application.

Fig. 3

MSCs were subcultured and plated in SlideFlasks and allowed to attach for 24 h before SlideFlasks were filled with culture medium and sealed. The treatment regimen for MSC’s involved 72 h of SMG (blue). LIVDT regimen consisted of one treatment cycle every 24 h during SMG treatment with each cycle consisting of 2 × 20 min LIV with an hour in between (yellow). LIVAT regimen was applied after 72 h SMG treatment period and consisted of 5 × 20 min LIV with an hour in between each (red). For LIV application, MSCs plated in SlideFlasks were placed in an LIV device constructed in the lab previous to this research. Vibrations were applied at peak magnitudes of 0.7 g at 90 Hz at room temperature. Control samples were treated the same but were not vibrated. For SMG application, MSCs plated in SlideFlasks were secured in lab custom-built clinostat inside the incubator. The clinostat subjected the MSCs to constant 15 RPM rotation simulated microgravity. After treatment, flasks were removed for immunofluorescence staining.