Jae Min Cha1,2, Yu-Shik Hwang3, Dong-Ku Kang4, Jun Lee5, Elana S Cooper6, Athanasios Mantalaris7. 1. Department of Mechatronics Engineering, College of Engineering, Incheon National University, 119 Academy-ro, Yeonsu-gu, Incheon, 22012, Republic of Korea. j.cha@inu.ac.kr. 2. 3D Stem Cell Bioengineering Laboratory, Research Institute for Engineering and Technology, Incheon National University, Incheon, 22012, Republic of Korea. j.cha@inu.ac.kr. 3. Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul, 02447, Republic of Korea. 4. Department of Chemistry, Research Institute of Basic Sciences, Incheon National University, Incheon, 22012, Republic of Korea. 5. Wonkwang Bone Regeneration Research Institute, Wonkwang University, Iksan, 570-749, Republic of Korea. 6. Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332, USA. 7. Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332, USA. sakis.mantalaris@gatech.edu.
Abstract
BACKGROUND: As stem cells are considered a promising cell source for tissue engineering, many culture strategies have been extensively studied to generate in vitro stem cell-based tissue constructs. However, most approaches using conventional tissue culture plates are limited by the lack of biological relevance in stem cell microenvironments required for neotissue formation. In this study, a novel perfusion rotating wall vessel (RWV) bioreactor was developed for mass-production of stem cell-based 3D tissue constructs. METHODS: An automated RWV bioreactor was fabricated, which is capable of controlling continuous medium perfusion, highly efficient gas exchange with surrounding air, as well as low-intensity pulsed ultrasound (LIPUS) stimulation. Embryonic stem cells encapsulated in alginate/gelatin hydrogel were cultured in the osteogenic medium by using our bioreactor system. Cellular viability, growth kinetics, and osteogenesis/mineralization were thoroughly evaluated, and culture media were profiled at real time. The in vivo efficacy was examined by a rabbit cranial defect model. RESULTS: Our bioreactor successfully maintained the optimal culture environments for stem cell proliferation, osteogenic differentiation, and mineralized tissue formation during the culture period. The mineralized tissue constructs produced by our bioreactor demonstrated higher void filling efficacy in the large bone defects compared to the group implanted with hydrogel beads only. In addition, the LIPUS modules mounted on our bioreactor successfully reached higher mineralization of the tissue constructs compared to the groups without LIPUS stimulation. CONCLUSION: This study suggests an effective biomanufacturing strategy for mass-production of implantable mineralized tissue constructs from stem cells that could be applicable to future clinical practice.
BACKGROUND: As stem cells are considered a promising cell source for tissue engineering, many culture strategies have been extensively studied to generate in vitro stem cell-based tissue constructs. However, most approaches using conventional tissue culture plates are limited by the lack of biological relevance in stem cell microenvironments required for neotissue formation. In this study, a novel perfusion rotating wall vessel (RWV) bioreactor was developed for mass-production of stem cell-based 3D tissue constructs. METHODS: An automated RWV bioreactor was fabricated, which is capable of controlling continuous medium perfusion, highly efficient gas exchange with surrounding air, as well as low-intensity pulsed ultrasound (LIPUS) stimulation. Embryonic stem cells encapsulated in alginate/gelatin hydrogel were cultured in the osteogenic medium by using our bioreactor system. Cellular viability, growth kinetics, and osteogenesis/mineralization were thoroughly evaluated, and culture media were profiled at real time. The in vivo efficacy was examined by a rabbit cranial defect model. RESULTS: Our bioreactor successfully maintained the optimal culture environments for stem cell proliferation, osteogenic differentiation, and mineralized tissue formation during the culture period. The mineralized tissue constructs produced by our bioreactor demonstrated higher void filling efficacy in the large bone defects compared to the group implanted with hydrogel beads only. In addition, the LIPUS modules mounted on our bioreactor successfully reached higher mineralization of the tissue constructs compared to the groups without LIPUS stimulation. CONCLUSION: This study suggests an effective biomanufacturing strategy for mass-production of implantable mineralized tissue constructs from stem cells that could be applicable to future clinical practice.
Authors: Stephen M Dang; Michael Kyba; Rita Perlingeiro; George Q Daley; Peter W Zandstra Journal: Biotechnol Bioeng Date: 2002-05-20 Impact factor: 4.530
Authors: Melissa L M Khoo; Leon R McQuade; Murray S R Smith; Justin G Lees; Kuldip S Sidhu; Bernard E Tuch Journal: Biol Reprod Date: 2005-08-03 Impact factor: 4.285
Authors: Mark J Perry; Laura K Parry; Victoria J Burton; Sabina Gheduzzi; Jon N Beresford; Victor F Humphrey; Tim M Skerry Journal: Med Eng Phys Date: 2008-05-20 Impact factor: 2.242