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. 2020 Jan 14;7:437. doi: 10.3389/fbioe.2019.00437

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Copyright © 2020 Shamsah, Cartmell, Richardson and Bosworth.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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(A) Schematic diagram of the cellular 3D ringed lamellar construct and demonstration of the topside region selected for confocal microscopy imaging. (B,C) Transverse confocal microscopy images demonstrating positive fluorescent staining of collagen type I (red) present in between the layers of the construct secreted by (D,E) annulus fibrosus (AF) cells (nuclei = blue) in vitro after 21 days in cell culture media. (F,G) Merged image. (Magnification B,D,F = x25, Scale bars = 1 mm). The white dashed square indicates the higher magnification imaging (x150) in (C,E,G) (Scale bar = 250 μm).