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. 2020 Dec 18;10(12):1696. doi: 10.3390/biom10121696

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Schematic representation of the experimental design. (A) Experimental groups. In 2D, adipose-derived stem cells were differentiated into osteogenic or adipogenic fate or maintained under standard conditions with and without electrical stimulation (ES). In 3D, cells were differentiated into osteogenic fate or kept in standard medium with and without exposure to ES. (B) After 4 days of pre-cultivation, medium was replaced by osteogenic, adipogenic, or fresh standard medium. XTT assays and immunocytochemical (ICC) staining against osteopontin (OPN) and osteocalcin (OCN) were performed at day d7, d14, and d21. Alkaline phosphatase (ALP) activity was assessed at d7, whereas Alizarin Red S staining, Oil Red O staining, live/dead assay and phalloidin staining were performed at d21.