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The major consideration for efficacy is that the mitochondria used in transplantation must be viable and intact to allow for uptake, internalization and efficacy.
The cells were incubated for 15 minutes at room temperature, in amber tubes, in the presence of the dyes to allow for uptake of Pr and adequate binding of Annexin V-FITC [18], [41], [42].
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Anaerobic conditions were simulated with the iTO977 model by constraining the oxygen uptake rate to zero, but also allowing for uptake of sterols to simulate that these compounds are supplied to the media for anaerobic growth.
While microbial folate catabolism pathways are not completely understood, it is clear that E. coli has a transporter that allows for uptake of pABA-glutamate, a folate breakdown product, and a hydrolase that can process pABA-glu into free pABA and glutamate [ 4].
In passive targeting, one takes advantage of the higher endocytic uptake of cancer cells and leaky vasculature around tumours which allow for higher uptake of NPs than in healthy tissues (Barreto et al. 2011), while in active targeting, the NPs are functionalised with specific molecules that interact with receptors known to be selectively present in tumour cells (Salvati et al. 2013).
On one hand it must allow for efficient uptake of nutrients and fluids, on the other hand it is a vital defence barrier between the milieu interior and the milieu exterior.
Cells were incubated 1 hour at 37°C to allow for dye uptake prior to fixation in PBS containing 4% paraformaldehyde.
Cells were washed to remove excess label, then incubated for 30 minutes at 37°C to allow for transferrin uptake.
For gentamicin protection assays, the multiplicity of infection was between 20 and 50 bacteria per cell to allow for sufficient uptake of bacteria.
For gentamicin protection assays, the multiplicity of infection was between 20 50 bacteria per cell to allow for sufficient uptake of bacteria.
However, extracellular and intracellular compartments are specified, and a minimal extracellular growth medium was applied to all models, along with necessary transport reactions that allow for its uptake.
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