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In these conditions, the metabolic network is free to absorb any medium or serum metabolites (at any rate), which include sugars, amino acids, several metabolic intermediates and short chain fatty acids.

Saliva has a number of different physical characteristics than, for example, cell culture medium or serum and assuming an ELISA which works well with serum samples will also do so with saliva potentially could lead to erroneous data and conclusions.

Therefore, in the present study, BM-hMSCs were cultivated in serum free growth medium or serum free growth medium with additional 4 or 6 mM Ca2+ for 3 weeks, following which, the proliferation and osteoblastic differentiation of these cells were evaluated.

The levels of HMGB1 in the culture medium or serum were determined by Western blotting analysis as previously described [7], [8], [12], [23].

The levels of TNF in the culture medium or serum were determined using commercial enzyme linked immunosorbant assay (ELISA) kits (Catalog no.

Murine cytokine antibody array (Cat. No. M0308003, RayBiotech Inc., Norcross, GA, USA), which detects 62 cytokines on one membrane, was used to determine the profile of cytokines in the culture medium or serum as previously described [12].

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OECs grown in astrocyte-conditioned medium or serum-free medium demonstrated only weak ERK1/2 phosphorylation (Fig. 5A, B).

bEnd5 cells were cultured in serum-free medium or serum-free medium containing 10 µg/ml FN in a µ-Slide I 0.4 Luer (Ibidi) over night.

Astrocyte-conditioned medium strongly increased invasiveness of melanoma cells compared with fibroblast-conditioned medium or serum-free medium (Fig. 4B and C).

Flow analysis showed that cytoplasmic B7-H4 was expressed in U251 cells cultured in either serum-free medium or serum-containing medium, whereas surface B7-H4 was difficult to detect in these two groups (data not shown).

According to light scattering analysis and analytical centrifugation, the particles used in this study were shown to be well separated, when dispersed in either serum-containing medium or serum-free medium.

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