Abstract
Mesenchymal stem cells (MSCs) hold considerable promise for regenerative medicine. Optimization of the seeding density of mononuclear cells (MNCs) improves the proliferative and differentiation potential of isolated MSCs. However, the underlying mechanism is unclear. We cultured human bone marrow MNCs at various seeding densities (4.0 × 104, 1.25 × 105, 2.5 × 105, 6.0 × 105, 1.25 × 106 cells/cm2) and examined MSC colony formation. At lower seeding densities (4.0 × 104, 1.25 × 105 cells/cm2), colonies varied in diameter and density, from dense to sparse. In these colonies, the proportion of highly proliferative MSCs increased over time. In contrast, lower proliferative MSCs enlarged more rapidly. Senescent cells were removed using a short detachment treatment. We found that these mechanisms increase the purity of highly proliferative MSCs. Thereafter, we compared MSCs isolated under optimized conditions with a higher density (1.25 × 106 cells/cm2). MSCs under optimized conditions exhibited significantly higher proliferative and differentiation potential into adipocytes and chondrocytes, except for osteocytes. We propose the following conditions to improve MSC quality: (1) optimizing MNC seeding density to form single-cell colonies; (2) adjusting incubation times to increase highly proliferative MSCs; and (3) establishing a detachment processing time that excludes senescent cells.
Reference40 articles.
1. Clinical trials with mesenchymal stem cells: An update;Squillaro;Cell Transplant.,2016
2. Mesenchymal stem or stromal cells: A review of clinical applications and manufacturing practices;Sharma;Transfusion,2014
3. Seong, J.M., Kim, B.-C., Park, J.-H., Kwon, K.I., Mantalaris, A., and Hwang, Y.-S. (2010). Stem cells in bone tissue engineering. Biomed. Mater., 5.
4. Human stem cells and articular cartilage tissue engineering;Stoltz;Curr. Pharm. Biotechnol.,2012
5. Intravenous infusion of mesenchymal stem cells promotes functional recovery in a rat model of chronic cerebral infarction;Namioka;J. Neurosurg.,2019
Cited by
1 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献