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. 2015 Oct 23;94(11):2784–2796. doi: 10.3382/ps/pev284

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© The Author 2015. Published by Oxford University Press on behalf of Poultry Science Association.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com.

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Figure 3. — Inhibition of calcium deposition in SPF-Layer (Lohmann LSL) MSCs undergoing osteogenic differentiation in the presence of high and low concentrations of 1,25(OH)₂D₃ in vitro. Mineralization was observed at all time points in cells undergoing osteogenic induction in the absence of exogenous 1,25(OH)₂D₃ (B, F, J). No mineralization is observed at Day 3 or 5 in the presence of 2.4nM, (C, G) or 24nM (D, H) 1,25(OH)₂D₃. (K) Only small foci of mineralization are observed in low 2.4nM 1,25(OH)₂D₃ at Day 7 of osteogenic differentiation A, E, I) MSCs grown in the absence of osteogenic inducing factors show no mineralization on any day examined. M) Quantitative analysis of calcium deposition expressed as area fraction (% of pixels). Values are mean ± SEM (n = 4 on all days examined). Comparisons marked with an asterisk (*) are significantly different (p < 0.05). Scale bar = 200μm. Abbreviations: CM, control media containing no exogenous 1,25(OH)₂D₃; IM, induction media; IM+2.4nM, induction media containing 2.4nM 1,25(OH)₂D₃; IM+24nM, induction media containing 24nM 1,25(OH)₂D₃; D, Day.