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. 2017 May 8;7:1548. doi: 10.1038/s41598-017-01528-3

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© The Author(s) 2017

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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ASC spheroid formation in HA gel. Schematic diagram of the preparation of ready-to-inject HA spheroids. HA powder was weighed and transferred to a syringe. Monolayer-cultured ASCs were trypsinized, counted, suspended in growth medium, aspirated into the syringe, and mixed with the powder, which was then allowed to dissolve for 10 min. The plunger was drawn back to assure air exchange through the needle. The needle cap then closed, and the syringe was incubated at 5% CO₂ and 37 °C. The gel slowly sank to the lower half of the syringe.