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. 2024 Apr 25;14:9495. doi: 10.1038/s41598-024-60221-4

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Development of 3D tendon cell culture system. (A) Three major steps of 3D tendon culture: cell extraction, cell growth in monolayer, and 3D culture. (B) The timeline of monolayer tendon cell culture after cell extraction. (C) The base of the 3D tendon cell culture consists of a growth channel area molded into 2% agarose using 3D-printed-mold and cylindrical anchors wrapped by hydrophilized PCL. Primary cells from the mouse tail tendon were seeded (2.5 × 10⁶ cells per construct) into the fibronectin-coated growth area to generate a 3D tendon construct. The dimension of growth area is 10 mm (length) × 1 mm (width) × 4 mm (depth). (D) The timeline of 3D tendon culture following monolayer culture (D, days after seeding of cells for 3D tendon culture; T, days after TGFβ1 treatment).