Subaqueous 3D stem cell spheroid levitation culture using anti-gravity bioreactor based on sound wave superposition

Biomaterials Research

Research Article

Ju-Ro Lee¹  Yoon Ki Joung²  Sungkwon Park³  Tae Il Lee⁴  Jeong-Kee Yoon⁵  Hyun Su Park⁶  Jung Hwan Park⁶  Yu-Jin Kim⁶  Suk Ho Bhang⁶  Jiyu Hyun⁶ 

[1] Center for Biomaterials, Biomedical Research Institute, Korea Institute of Science and Technology, 02792, Seoul, Republic of Korea;Center for Biomaterials, Biomedical Research Institute, Korea Institute of Science and Technology, 02792, Seoul, Republic of Korea;Division of Bio-Medical Science and Technology, University of Science and Technology, Republic of Korea, 02792, Seoul, Republic of Korea;Department of Food Science and Biotechnology, College of Life Science, Sejong University, 05006, Seoul, Korea;Department of Materials Science and Engineering, Gachon University, Gyeonggi-Do, 13120, Seongnam-Si, Republic of Korea;Department of Systems Biotechnology, Chung-Ang University, Gyeonggi-Do, 17540, Anseong-Si, Republic of Korea;School of Chemical Engineering, Sungkyunkwan University, 16419, Suwon, Republic of Korea;

关键词: Acoustic levitation;    Anti-gravity bioreactor;    Human mesenchymal stem cell spheroid;    Hindlimb ischemia;   

DOI  :  10.1186/s40824-023-00383-w

 received in 2023-01-30, accepted in 2023-04-19,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundRecently, various studies have revealed that 3D cell spheroids have several advantages over 2D cells in stem cell culture. However, conventional 3D spheroid culture methods have some disadvantages and limitations such as time required for spheroid formation and complexity of the experimental process. Here, we used acoustic levitation as cell culture platform to overcome the limitation of conventional 3D culture methods.MethodsIn our anti-gravity bioreactor, continuous standing sonic waves created pressure field for 3D culture of human mesenchymal stem cells (hMSCs). hMSCs were trapped and aggerated in pressure field and consequently formed spheroids. The structure, viability, gene and protein expression of spheroids formed in the anti-gravity bioreactor were analyzed by electron microscope, immunostaining, polymerase chain reaction, and western blot. We injected hMSC spheroids fabricated by anti-gravity bioreactor into the mouse hindlimb ischemia model. Limb salvage was quantified to evaluate therapeutic efficacy of hMSC spheroids.ResultsThe acoustic levitation in anti-gravity bioreactor made spheroids faster and more compact compared to the conventional hanging drop method, which resulted in the upregulation of angiogenic paracrine factors of hMSCs, such as vascular endothelial growth factor and angiopoietin 2. Injected hMSCs spheroids cultured in the anti-gravity bioreactor exhibited improved therapeutic efficacy, including the degree of limb salvage, capillary formation, and attenuation of fibrosis and inflammation, for mouse hindlimb ischemia model compared to spheroids formed by the conventional hanging drop method.ConclusionOur stem cell culture system using acoustic levitation will be proposed as a new platform for the future 3D cell culture system.Graphical Abstract

【 授权许可】

CC BY   
© The Author(s) 2023

【 图 表 】

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