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. 2019 Jun 12;5(6):eaaw4991. doi: 10.1126/sciadv.aaw4991

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Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 4 — (A) Schematic diagram of coculture design indicating the placement of differentiating hMSCs on Col-GAG or MC-GAG within Transwell insert and lower chamber consisting of primary pre-osteoclasts cultured on a plate coated with calcium phosphate to allow for the detection of resorptive pit activity. (B) WST-1 assays of hMSCs in single culture (hMSC only) or cocultured (control hMSC/OC and AdOPG hMSC/OC, respectively) on Col-GAG or MC-GAG for 21 days. Empty cell-free scaffolds cocultured with osteoclasts shown for control (empty scaffolds/OC). (C) OPG ELISA of hMSC/OC coculture medium (days 3, 7, 10, and 14) with control and AdOPG-transduced hMSCs on Col-GAG and MC-GAG. Differentiated osteoclast-only (OC only) culture shown at the left as a control. (D) Representative μCT images and (E) quantitative analysis of empty scaffold (empty), hMSCs without osteoclasts (hMSC only), control hMSCs cocultured with osteoclasts, or AdOPG-transduced hMSCs cocultured with osteoclasts on Col-GAG or MC-GAG for 21 days. (F) WST-1 assays of primary pre-osteoclasts in single culture (OC only), cocultured with empty scaffolds, or cocultured with control or AdOPG-transduced hMSCs (control hMSC and AdOPG hMSC, respectively) in medium supplemented with RANKL and M-CSF on Col-GAG or MC-GAG for 14 days. (G) Quantitative analysis of pit assays as the percentage of total area of well in differentiated osteoclasts without hMSCs (OC only) and osteoclasts cocultured with Col-GAG and MC-GAG as empty scaffolds, scaffolds with control hMSCs, and scaffolds with AdOPG-transduced hMSCs. Significant post hoc comparisons following ANOVA indicated with P values.