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. 2022 Nov 18;23(22):14349. doi: 10.3390/ijms232214349

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© 2022 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 (https://creativecommons.org/licenses/by/4.0/).

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Spheroid formation and MSCs characterization. (A) Chronological record of 3D-MSCs compaction. Images were captured at 0, 1, 2, 4, 12, 24, 48, and 72 h after 3D-MSCs seeding on 96-well agarose-coated plates at 4 × magnification. (B) Scanning electron microscopy images. From left to right the magnifications are 350, 2000, and 4000 ×. (C) CD44, CD73, CD90, and CD105 expression on 2D-MSCs (2D) and 3D-MSCs (3D). (D) Representative histograms of one MSC sample evaluated. Red color corresponds with 2D-MSCs, while blue color corresponds with 3D-MSCs. (E) Percentages of apoptotic (annexin V+) 2D and 3D-MSCs evaluated after their dissociation with collagenase IA. (F) Representative flow cytometric dot plots showing expressions of PI and Annexin V in 2D and 3D-MSCs. (G) LDH release by 2D and 3D-MSCs. ** p < 0.01; *** p < 0.001; **** p < 0.0001.