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Analyzing the components in our defined medium preparations, we identified three major sources of xeno material: BSA as a component of various media formulations, geltrex (which was used as a substrate in several stages of the process) and PA6-CM.
The fermentation medium preparations are described below.
Each of these medium preparations was measured using a LabMaster- a w meter (Novasina AG, Lanchen, Switzerland).
It is difficult to directly compare the results obtained in different studies, because different osteoclastogenic assays as well as conditioned medium preparations were used.
Interestingly, although the a w measurements of all three medium preparations were nearly identical, no upregulated genes were shared between all three conditions after 1 h of exposure.
All F1 medium preparations were supplemented with the following per liter: 20 mg histidine; 100 mg leucine; 30 mg lysine; 20 mg methionine; and 20 mg uracil.
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Embryos that had progressed to at least the 2-cell stage were cultured in 20 μl drops of KSOM (Lawitts and Biggers, 1993) (prepared according to the Jackson Laboratory medium preparation protocol) covered with mineral oil (FertiPro) in a humidified chamber at 37°C supplemented with 5% CO2.
Liquid seed medium inoculation was performed using conventional liquid seed medium preparation procedure.
Being rich in sucrose, sugarcane molasses represents a low-cost alternative carbon source suitable for medium preparation (Miljković et al. 2016).
The nutrients used in medium preparation are not recycled during the MEOR process; hence, the cost involved would affect the selection of optimized condition.
Mechanical property index Imp could well comprehensively characterize physical properties of medium, which could be useful for guiding initial culture medium preparation and SSF process control.
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