Effect of Dynamic Culture and Periodic Compression on Human Mesenchymal Stem Cell Proliferation and Chondrogenesis
- 936 Downloads
We have recently developed a bioreactor that can apply both shear and compressive forces to engineered tissues in dynamic culture. In our system, alginate hydrogel beads with encapsulated human mesenchymal stem cells (hMSCs) were cultured under different dynamic conditions while subjected to periodic, compressive force. A customized pressure sensor was developed to track the pressure fluctuations when shear forces and compressive forces were applied. Compared to static culture, dynamic culture can maintain a higher cell population throughout the study. With the application of only shear stress, qRT-PCR and immunohistochemistry revealed that hMSCs experienced less chondrogenic differentiation than the static group. The second study showed that chondrogenic differentiation was enhanced by additional mechanical compression. After 14 days, alcian blue staining showed more extracellular matrix formed in the compression group. The upregulation of the positive chondrogenic markers such as Sox 9, aggrecan, and type II collagen were demonstrated by qPCR. Our bioreactor provides a novel approach to apply mechanical forces to engineered cartilage. Results suggest that a combination of dynamic culture with proper mechanical stimulation may promote efficient progenitor cell expansion in vitro, thereby allowing the culture of clinically relevant articular chondrocytes for the treatment of articular cartilage defects.
KeywordsMesenchymal stem cell Chondrogenesis Differentiation Compression Cartilage Dynamic culture
This study was funded by the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health (R01 AR061460) as well as by the National Science Foundation (CBET 1264517). This work was also funded by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry (Grant number: 230303). The authors thank Feng Gao from Cornell University for his help on data processing and Dr. Hannah B. Baker for reviewing the manuscript.
- 6.Felson, D. T., R. C. Lawrence, P. A. Dieppe, R. Hirsch, C. G. Helmick, J. M. Jordan, R. S. Kington, N. E. Lane, M. C. Nevitt, Y. Q. Zhang, M. Sowers, T. McAlindon, T. D. Spector, A. R. Poole, S. Z. Yanovski, G. Ateshian, L. Sharma, J. A. Buckwalter, K. D. Brandt, and J. F. Fries. Osteoarthritis: new insights. Part 1: The disease and its risk factors. Ann. Intern. Med. 133:635–646, 2000.CrossRefPubMedGoogle Scholar
- 19.Longobardi, L., L. O’Rear, S. Aakula, B. Johnstone, K. Shimer, A. Chytil, W. A. Horton, H. L. Moses, and A. Spagnoli. Effect of IGF-I in the chondrogenesis of bone marrow mesenchymal stem cells in the presence or absence of TGF-beta signaling. J. Bone Miner. Res. 21:626–636, 2006.CrossRefPubMedGoogle Scholar
- 27.Smith R. L., D. R. Carter and D. J. Schurman. Pressure and shear differentially alter human articular chondrocyte metabolism: a review. Clin. Orthop. Relat. Res. S89–S95, 2004.Google Scholar
- 28.Takahashi, I., G. H. Nuckolls, K. Takahashi, O. Tanaka, I. Semba, R. Dashner, L. Shum, and H. C. Slavkin. Compressive force promotes sox9, type II collagen and aggrecan and inhibits IL-1beta expression resulting in chondrogenesis in mouse embryonic limb bud mesenchymal cells. J. Cell Sci. 111(Pt 14):2067–2076, 1998.PubMedGoogle Scholar