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In order to compare this method with previously published results, we also exposed animals to a train of mechanostimulatory events by repeatedly exposing their cultivation dish to a non-directional mechanical shock (dropping) and monitoring their reversal magnitude response (Fig. 7A, bottom right).
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The cell lines were cultivated in Nunc (Thermo Fisher Scientific) cultivation dishes, flasks, and plates in a humidified incubator at 37°C in an atmosphere of 5% CO2.
Multiple animals (20 30) were placed on 6 cm cultivation dishes inside a secondary plastic container at 25°C, and then dropped ∼4 cm once a minute for 15 minutes.
As a control, we also raised animals in an environment deprived of mechanical stimulation, buffering cultivation dishes from vibration using sorbothane.
We raised newly hatched animals in a regimented, mechanically stimulated environment by tapping their cultivation dishes once a minute for 15 minutes, repeating at one-hour intervals 7 times during the early larval period (Fig. 3A).
Static cultivation in a culture dish was also carried out.
When the cells were treated with a 200 MPa, the cells were floating on the culture dish after the cultivation at 3 and 24 hours, and the dehydrogenate activity was completely suppressed.
The resulting co-cultivation dishes allowed the growth of the plantlets over 10 days in the presence of the bacteria at 22 °C under 94 μmol m−2 s−1 in the PAR (LED lighting) with a 20-h photoperiod.
Cell-free MSC conditioned medium (CM) was collected from 2 × 10 cells plated on a 35 mm culture dish after 48 h of cultivation in high-glucose medium and filtered through 0.45 μm filters.
After these manipulations, all papillae were attached to the culture dish, and after 2 days of cultivation, the typical spindle-shaped fibrocyte-like cells grew from the explants).
Primary WT and P1−/− myoblasts were isolated from neonatal skeletal limb muscles of WT and isoform P1 knockout mice by spreading cells onto collagen-coated culture dishes, and cultivation in 20% FCS and basic fibroblast growth factor-supplemented growth medium (20).
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