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Updated: Jun 17, 2026
Fabrication of 3D Cardiac Microtissue Arrays using Human iPSC-Derived Cardiomyocytes, Cardiac Fibroblasts, and Endothelial Cells
Published on: March 14, 2021
Micro and nano-scale in vitro 3D culture system for cardiac stem cells
Hossein Hosseinkhani1, Mohsen Hosseinkhani, Shunji Hattori
1International Research Institute for Integrated Medical Sciences (IREIIMS), Tokyo Women's Medical University, Tokyo 162-8666, Japan. hossein@imcir.twmu.ac.jp
Insights
Developing a novel 3D culture system using collagen-PGA composites significantly enhanced cardiac stem cell (CSC) attachment and proliferation for heart regeneration. This biomaterial scaffold supports CSC expansion for potential therapeutic applications.
Area of Science:
- Biomaterials Science
- Regenerative Medicine
- Cardiovascular Research
Background:
- Restoring function in damaged hearts is challenging despite advances in cardiovascular disease prevention.
- Cardiac stem cells (CSCs) offer regenerative potential but face hurdles in generating sufficient numbers for clinical use.
- Existing methods struggle to produce adequate quantities of phenotypically consistent CSCs for therapy.
Purpose of the Study:
- To develop a three-dimensional (3D) culture system using micro- and nano-scaled materials for enhanced cardiac stem cell (CSC) expansion.
- To evaluate the efficacy of collagen-poly(glycolic acid) (PGA) composite scaffolds in supporting CSC attachment, proliferation, and phenotype.
- To compare the performance of the 3D culture system with traditional 2D culture and static 3D culture methods.
Main Methods:
- Fabrication of electrospun poly(glycolic acid) (PGA) nanofibers and their incorporation into collagen sponges to create composite scaffolds.
- Characterization of scaffold structure and mechanical properties using scanning electron microscopy (SEM) and compressive strength analysis.
- Assessment of cardiac stem cell (CSC) attachment and proliferation on collagen-PGA scaffolds in a bioreactor perfusion system versus static and 2D cultures.
Main Results:
- PGA nanofibers significantly improved the compressive strength of collagen sponges.
- Collagen-PGA composite scaffolds, particularly with 6 mg of PGA, demonstrated enhanced CSC attachment compared to controls.
- The 3D culture system with bioreactor perfusion significantly improved CSC attachment and proliferation over static 3D and 2D cultures.
Conclusions:
- The combination of micro- and nano-scaled materials in a 3D culture system shows great promise for culturing stem cells.
- Collagen-PGA composite scaffolds provide a supportive microenvironment for cardiac stem cell expansion.
- This advanced 3D culture approach is a promising strategy for generating sufficient CSCs for regenerative medicine applications in cardiovascular repair.
Abstract:
Despite the success to prevent or limit cardiovascular diseases, the restoration of the function of a damaged heart remains a formidable challenge. Cardiac stem cells (CSCs), with the capacity to differentiate into cardiomyocytes, hold great potential as a source of cells for regenerative medicine. A major challenge facing the clinical application of differentiated CSCs, however, is theability to generate sufficient numbers of cells with the desired phenotype. We previously established cell lines of CSCs using a c-kit antibody from adult rat hearts for use in regenerative medicine. C-kit -positive cardiac cells are well recognized as CSCs and have the potential to differentiate into cardiomyocytes. Here, before implant these cells in vivo, we first developed three-dimensional culture system (3D) using micro- and nano-scaled material. Sheets of poly(glycolic acid) (PGA) were fabricated by electrospinning. Composites of collagen-PGA were prepared that contained 0, 1.5, 3 or 6 mg of electrospun PGA nanofibers. The nanofibers were added as a sheet that formed a layer within the collagen sponge. The sponges were freeze-dried and then dehydrothermally crosslinked. A scanning electron microscopy (SEM)-based analysis of the surface of the sponges demonstrated a uniform collagenous structure regardless of the amount of PGA nanofibres included. The PGA nanofibers significantly enhanced the compressive strength of the collagen sponge. More CSCs attached to the collagen sponge incorporating 6 mg of PGA nanofibers than the sponge without PGA nanofibers. The attachment and proliferation of CSCs in the 3D culture was enhanced by incubation in a bioreactor perfusion system compared with 3D static and two-dimensional (2D; i.e. tissue culture plates) culture systems. The use of micro- and nano-scale materials in the fabrication of composites together with a 3D culture system is a very promising way to promote the culture of stem cells. (c) 2009 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.
