Author:
Park Jung Hwan,Lee Ju-Ro,Park Sungkwon,Kim Yu-Jin,Yoon Jeong-Kee,Park Hyun Su,Hyun Jiyu,Joung Yoon Ki,Lee Tae Il,Bhang Suk Ho
Abstract
Abstract
Background
Recently, 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.
Methods
In 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.
Results
The 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.
Conclusion
Our stem cell culture system using acoustic levitation will be proposed as a new platform for the future 3D cell culture system.
Graphical Abstract
Funder
National Research Foundation of Korea
Korean Fund for Regenerative Medicine
Ministry of Trade, Industry and Energy
Korea Institute of Science and Technology
Publisher
American Association for the Advancement of Science (AAAS)
Subject
Biomedical Engineering,Biomaterials,Medicine (miscellaneous),Ceramics and Composites
Cited by
1 articles.
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