Multilineage Differentiation of hASCs

Adipogenic, osteogenic and chondrogenic differentiation were performed to evaluate the multidifferentiation potential and a possible impact of VSOPs labeling. The hASCs were labeled with 1.5 mM VSOPs, the highest concentration used for the experiments. In the literature, an impairment of the differentiation potential of MSCs as a function of IONPs concentration is described [51 (link),52 (link)]. Defined media were used to induce differentiation. For adipogenic differentiation, EM-DMEM supplemented with 10 µM dexamethasone, 1 µg/mL insulin, 100 µM indomethacin and 500 µM 1-methyl-3-isobutylxanthine was used based on a modification [65 (link)] of the protocol of Pittenger et al. [66 (link)]. The osteogenic differentiation medium consists of EM-DMEM with 100 nM dexamethasone, 10 mM ß-glycerophosphate and 50 µg/mL ascorbic acid according to Jaiswal et al. [67 (link)]. Chondrogenic differentiation was induced with DMEM plus 1% P/S and 100 nM dexamethasone, 100 µg/mL sodium pyruvate, 50 µg/mL ascorbate-2-phosphate, 40 µg/mL proline, ITS-plus (Sigma-Aldrich Chemie GmbH) and 10 ng/mL TGF-ß3 (LONZA, Basel, Switzerland).