1Department of Chemical and Biomolecular Engineering, Seoul National University of Science and Technology, 232 Gongneung-ro, Nowon-gu, Seoul, 11811 Republic of Korea.
2Convergence Institute of Biomedical Engineering and Biomaterials, Seoul National University of Science and Technology, 232 Gongneung-ro, Nowon-gu, Seoul, 11811 Republic of Korea.
Tissue Eng Regen Med. 2019 Jan 28;16(1):19-28. doi: 10.1007/s13770-018-00174-8. eCollection 2019 Feb.
Tissue engineering is a multidisciplinary field which attracted much attention in recent years. One of the most important issue in tissue engineering is how to obtain high cell numbers and tissue regeneration while maintaining appropriate cellular characteristics for restoring damaged or dysfunctional body tissues and organs. These demands can be achieved by the use of three dimensional (3D) dynamic cultures of cells combined with cell-adhesive micro-carriers.
In this study, human mesenchymal stem cells (hMSCs) were cultured in a wave-bioreactor system for up to 100 days, after seeding on Cultisphere-S porous gelatin micro-carriers. Cell counting was performed at the time points of 7, 12, 17, 31 days and compared to those of hMSCs cultured under static condition. Higher growth and proliferation rates was achieved in wave-type dynamic culture, when cell culture continued to day 31. A scanning electron microscope (SEM) photographs, both live and dead and MTT assays were taken to confirm the survival and distribution of cells on porous gelatin micro-carrier surfaces. The results of histological stains such as hematoxylin and eosin, Masson's trichrome, Alcian blue and Alizarin red S also showed improved proliferation and tissue regeneration of hMSCs on porous gelatin micro-carriers.
The experimental results demonstrated the effect and importance of both micro-carriers and bioreactor in hMSC expansion on cell proliferation and migration as well as extracellular matrix formation on the superficial and pore surfaces of the porous gelatin micro-carriers, and then their inter-connections, leading to tissue regeneration.
组织工程是一个多学科领域，近年来受到了广泛关注。组织工程中最重要的问题之一是如何在保持适当细胞特性的情况下获得高细胞数量和组织再生，以修复受损或功能失调的身体组织和器官。这一需求可以通过使用三维（3D）动态细胞培养与细胞黏附微载体相结合来实现。
在这项研究中，人骨髓间充质干细胞（hMSCs）在波生物反应器系统中培养，时间长达 100 天，在接种 Cultisphere-S 多孔明胶微载体后。在第 7、12、17 和 31 天进行细胞计数，并与静态培养的 hMSCs 进行比较。在波型动态培养中，当细胞培养持续到第 31 天时，实现了更高的生长和增殖率。通过扫描电子显微镜（SEM）照片、活细胞和死细胞以及 MTT 检测，证实了细胞在多孔明胶微载体表面的存活和分布。苏木精-伊红、马松三色、阿利新蓝和茜素红 S 等组织学染色的结果也表明，多孔明胶微载体上 hMSCs 的增殖和组织再生得到了改善。
实验结果表明，微载体和生物反应器在 hMSC 增殖、细胞迁移以及细胞外基质在多孔明胶微载体表面和孔内表面的形成及其相互连接，从而导致组织再生方面的作用和重要性。
