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Thus, scaffold\u2010free tissue\u2010engineered structures can be a promising solution to overcome the issues associated with classical scaffold\u2010based materials in clinical transplantation. The present study seeks to optimize the culture conditions and cell combinations used to generate scaffold\u2010free structures using a Bio\u20103D printing system. Human cartilage cells, human fibroblasts, human umbilical vein endothelial cells, and human mesenchymal stem cells from bone marrow are aggregated into spheroids and placed into a Bio\u20103D printing system with dedicated needles positioned according to 3D configuration data, to develop scaffold\u2010free trachea\u2010like tubes. Culturing the Bio\u20103D\u2010printed structures with proper flow of specific medium in a bioreactor facilitates the rearrangement and self\u2010organization of cells, improving physical strength and tissue function. The Bio\u20103D\u2010printed tissue forms small\u2010diameter trachea\u2010like tubes that are implanted into rats with the support of catheters. It is confirmed that the tubes are viable in vivo and that the tracheal epithelium and capillaries proliferate. This tissue\u2010engineered, scaffold\u2010free, tubular structure can represent a significant step toward clinical application of bioengineered organs.<\/jats:p>","DOI":"10.1002\/adhm.201800983","type":"journal-article","created":{"date-parts":[[2019,1,11]],"date-time":"2019-01-11T08:11:01Z","timestamp":1547194261000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":89,"title":["Replacement of Rat Tracheas by Layered, Trachea\u2010Like, Scaffold\u2010Free Structures of Human Cells Using a Bio\u20103D Printing System"],"prefix":"10.1002","volume":"8","author":[{"given":"Ryusuke","family":"Machino","sequence":"first","affiliation":[{"name":"Department of Surgical Oncology Nagasaki University Graduate School of Biomedical Sciences  Nagasaki 852\u20108501 Japan"},{"name":"Medical\u2010Engineering Hybrid Professional Development Center Nagasaki University Graduate School of Biomedical Sciences  Nagasaki 8528501 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