Finally, to confirm the differentiation of TSCs into osteocytes in osteogenic medium, induction of Runx2, an osteoblast specific gene, was examined by qRT-PCR.
From this liquid culture, two spots of 5 μl were made on solid LB medium; induction of colicin production was conducted adding mitomycin C in the solid LB medium at a final concentration of 0.5 μg/ml.
The measurements in the regimes of weak and medium induction reflect the activity of the promoter, which is regulated by the repressors (LacI and AraC) and activators (IPTG and Arabinose), since the binding and unbinding of these molecules to the promoter is a process whose speed is orders of magnitude faster than the process of initiation.
Nine factors were selected (the concentration of glycerol, tryptone, yeast extract, PBS buffer, and MOPS in the medium, induction occasion, the concentration of IPTG, inoculation, loading volume), each of which was coded with two levels (Table 1).
As expected, UC-MSCs and ASCs conditioned medium induction led to significant overexpression of pro-apoptotic proteins such as Bad, Bax, Bim, Bid, etc. and significant downregulation of anti-apoptotic proteins such as Bcl-2, Survivin and XIAP.
All strains were grown in a defined sorbitol medium when heterologous lysozyme expression was not required and in a defined sorbitol methanol medium for induction of expression.
It has been observed earlier that in some cases withdrawal of auxins from the culture medium promotes induction of somatic embryogenesis in calli (Gutiérrez-Mora et al. 2012; Lema-Rumińska et al. 2013).
The C-105 cells were cultivated in urate containing medium for induction of uricase.
Interestingly, we found a strong effect of MSC conditioned medium on induction of differentiation of U251 glioma cells.
To avoid any interference of FCS-derived serum factors, cells were cultured overnight in UC medium before induction of apoptosis.
After 40 d of culture, all explants were transferred to fresh MSB medium for induction of embryogenic calli (ECs).
