We used flow cytometry to analyze cell surface
marker expressions by the ASCs culture
prior to ZnO-NPs treatment. The cells were
characterized according to a set of cell surface
markers characteristic for ASCs that included
cluster of differentiation (CD) CD73 (sc-14682,
Santa Cruz Biotechnology Inc., USA), CD105
(sc-20632, Santa Cruz Biotechnology Inc.,
USA), CD45 (sc-25590, Santa Cruz Biotechnology
Inc., USA) and CD34 (sc-7045, Santa
Cruz Biotechnology Inc., USA).The differentiation
potentials of the ASCs were checked in
specific media. For adipocyte differentiation,
cells were cultured in 1 μmol/l dexamethasone
(D4902, Sigma, USA), 60 μmol/l indomethacin
(I7378, Sigma, USA), and 450 μl 3-isobutyl-
1-methylxanthine (I5879, Sigma, USA). Adipocytes
were characterized by oil red-O (O0652,
Sigma, USA) staining under a light microscope.
For differentiation into osteoblasts, culture medium
was supplemented with 0.1 μmol/l dexamethasone,
10 mmol/l β-glycerophosphate
(G9422, Sigma, USA) and 60 mmol/l ascorbate
(A0157, Sigma, USA). Osteoblasts were characterized
by alizarin red (A5533, Sigma, USA)
staining and microscopic examination (16 (link)).
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