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The hydrophilic analytes in the then-organic medium were subsequently extracted by the HILIC trap column.
Aliquots of medium were subsequently removed and analyzed spectrophotometrically at a wavelength of 560 nm.
ECs incubated with the respective medium were subsequently obtained after 24 h and 48 h under normal gravity incubation and numbers were calculated.
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The culture medium was subsequently discarded and replaced with 150 μL of DMSO.
Fresh medium was subsequently added, and the cells were shifted to 37 °C to allow synchronous infection.
The filtered TSB medium was subsequently incubated at 30°C for 14 days.
M9-based culture medium was subsequently optimized for the enzyme production by orthogonal experimental design in shake flasks.
The culture medium was subsequently replaced with specific inductive media to determine the adipogenic, osteogenic, and chondrogenic differentiation potential as described previously (Marchal et al. 2012).
Whole medium was subsequently replaced weekly.
The medium was subsequently changed every other day.
The medium was subsequently aspirated and cells were quickly washed twice with ice-cold Hank's balanced salt solution (HBSS) and harvested with tripsin-EDTA.
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