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Currently, there is no adequate solution for patients requiring the resection of more than half of the length of their trachea. In this study, the best conditions for combination of three different cell types from the respiratory airway system were investigated to develop a functional ciliated and pre\u2010vascularized mucosal substitute in vitro. Primary human fibroblasts were combined with respiratory epithelial cells and endothelial cells. As scaffolds, fibrin gel and agarose\u2010type I collagen blends were used and cultured with different medium compositions to optimize both vascularization and differentiation of the respiratory epithelium. A mixture of endothelial growth medium and epithelial differentiation medium was shown to optimize both vascularization and epithelial growth and differentiation. After 28\u2009days of co\u2010culture, significantly increased formation of capillary\u2010like structures was observed in fibrin gels with more than three times higher structure volumes compared to agarose\u2013collagen gels. After 35\u2009days, epithelial differentiation into a pseudostratified epithelium with typical marker expression was improved on fibrin gels. While cilia formation was shown on both scaffolds, a higher number of ciliated cells and longer cilia were observed on fibrin gels. The data elucidate the important interplay of co\u2010culture parameters and their impact on vascularization as well as epithelium development and provide a basis for development of functional three\u2010dimensional airway constructs.<\/jats:p>","DOI":"10.1002\/jbm.a.36718","type":"journal-article","created":{"date-parts":[[2019,5,13]],"date-time":"2019-05-13T14:50:21Z","timestamp":1557759021000},"page":"2053-2062","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Combination of vascularization and cilia formation for three\u2010dimensional airway tissue engineering"],"prefix":"10.1002","volume":"107","author":[{"given":"Franziska","family":"Kreimendahl","sequence":"first","affiliation":[{"name":"Department of Biohybrid and Medical Textiles (BioTex), AME \u2010 Institute of Applied Medical Engineering, Helmholtz Institute RWTH Aachen University  Aachen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sina","family":"Ossenbrink","sequence":"additional","affiliation":[{"name":"Department of Biohybrid and Medical Textiles (BioTex), AME \u2010 Institute of Applied Medical Engineering, Helmholtz Institute RWTH Aachen University  Aachen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marius","family":"K\u00f6pf","sequence":"additional","affiliation":[{"name":"Department of Dental Materials and Biomaterials Research RWTH Aachen University Hospital  Aachen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Westhofen","sequence":"additional","affiliation":[{"name":"Clinic for Otorhinolaryngology and Plastic Surgery of the Head and Throat RWTH Aachen University Hospital  Aachen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Schmitz\u2010Rode","sequence":"additional","affiliation":[{"name":"Department of Biohybrid and Medical Textiles (BioTex), AME \u2010 Institute of Applied Medical Engineering, Helmholtz Institute RWTH Aachen University  Aachen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Horst","family":"Fischer","sequence":"additional","affiliation":[{"name":"Department of Dental Materials and Biomaterials Research RWTH Aachen University Hospital  Aachen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Jockenhoevel","sequence":"additional","affiliation":[{"name":"Department of Biohybrid and Medical Textiles (BioTex), AME \u2010 Institute of Applied Medical Engineering, Helmholtz Institute RWTH Aachen University  Aachen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anja L.","family":"Thiebes","sequence":"additional","affiliation":[{"name":"Department of Biohybrid and Medical Textiles (BioTex), AME \u2010 Institute of Applied Medical Engineering, Helmholtz Institute RWTH Aachen University  Aachen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2019,5,21]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/s40248-016-0046-3"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/0962-8924(96)84935-X"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0177(199703)208:3<398::AID-AJA10>3.0.CO;2-X"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10439-011-0437-8"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1064829"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa0810653"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1097\/MCP.0b013e3283539673"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.02211-2017"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bioeng.2.1.227"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1165\/ajrcmb.20.2.3062"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1089\/ten.2005.11.257"},{"key":"e_1_2_7_13_1","first-page":"L347","article-title":"Culture and transformation of human airway epithelial cells","volume":"268","author":"Gruenert D. 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