A Novel Stem Cell Tag-Less Sorting Method
Tóm tắt
Growing interest in stem cell research has led to the development of a number of new methods for isolation. The lack of homogeneity in stem cell preparation blurs standardization, which however is recommended for successful applications. Among stem cells, mesenchymal stem cells (MSCs) are promising candidates for cell therapy applications. This paper presents a fractionation protocol based on a tag-less, flow-assisted method of purifying, distinguishing and sorting MSCs. The protocol entails a suspension of cells in a transport fluid being injected into a ribbon-like capillary device by continuous flow. In a relatively short time (about 30 min) sorted cells are collected. The protocol has been applied to the improvement of MSC isolation, with a specific view to reducing cell manipulation operations, keeping instrumental simplicity and increasing analytical information for cell characterization. Applications such as MSC purification from epithelial contaminants, MSC characterization from various human sources and sorting of MSC subpopulations with high differentiation potential are described. The low cost, full biocompatibility and scale-up potential of the protocol presented could make the procedure attractive for stem cell selection.
Tài liệu tham khảo
Wagner, W., Wein, F., Seckinger, A., Frankhauser, M., Wirkner, U., Krause, U., et al. (2005). Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. Experimental Hematology, 33, 1402–1416.
Tong, J., Hoffman, R., Siena, S., Srour, E. F., Bregini, M., & Gianni, A. M. (1994). Characterization and quantification of primitive hematopoietic progenitor cells present in peripheral blood autograft. Experimental Hematology, 22, 1016–1024.
Lucas, A., Lepage, F., & Cardot, P. (2000). Cell separations. In M. E. Schimpf, K. Caldwell & J. C. Giddings (Eds.), Field-flow fractionation hand-book, chapter 29 (pp. 471–486). New York: Wiley-Interscience.
Reschiglian, P., Zattoni, A., Roda, B., Michelini, E., & Roda, A. (2005). Field-flow fractionation and biotechnology. Trends in Biotechnology, 23, 475–83.
Bernard, A., Paulet, B., Colin, V., & Cardot, P. (1995). Red Blood cell separations by gravitational field-flow fractionation: instrumentation & application. Trends in Analytical Chemistry, 14, 266–273.
Roda, B., Reschiglian, P., Zattoni, A., Tazzari, P. L., Buzzi, M., Ricci, F., et al. (2008). Human lymphocyte sorting by gravitational field-flow fractionation. Analytical and Bioanalytical Chemistry, 392, 137–145.
Roda, B., Reschiglian, P., Alviano, F., Lanzoni, G., Bagnara, G. P., Ricci, F., et al. (2009). Gravitational field-flow fractionation of human hemopoietic stem cells. Journal of Chromatography A, . doi:10.1016/j.chroma.2009.07.024.
Parolini, O., Alviano, F., Bagnara, G. P., Bilic, G., Bühring, H. J., Evangelista, M., et al. (2008). Concise review: isolation and characterization of cells from human term placenta: outcome of the first international Workshop on Placenta Derived Stem Cells. Stem Cells, 26, 300–311.
Marchionni, C., Bonsi, L., Alviano, F., Lanzoni, L., Di Tullio, A., Costa, R. (2009). Angiogenic potential of human dental pulp stromal (stem) cells. International Journal of Immunopathology and Pharmacology, in press.
Pittenger, M. F., Mackay, A. M., Beck, S. C., Jaiswal, R. K., Douglas, R., Mosca, J. D., et al. (1999). Multilineage potential of adult human mesenchymal stem cells. Science, 284, 143–147.
Alviano, F., Fossati, V., Marchionni, C., Arpinati, M., Bonsi, L., Franchina, M., et al. (2007). Term Amniotic membrane is a high throughput source for multipotent Mesenchymal Stem Cells with the ability to differentiate into endothelial cells in vitro. BMC Developmental Biology, 21, 7–11.
Pasquinelli, G., Tazzari, P. L., Vaselli, C., Foroni, L., Buzzi, M., Storci, G., et al. (2007). Thoracic aortas from multiorgan donors are suitable for obtaining resident angiogenic mesenchymal stromal cells. Stem Cells, 2007(25), 1627–1634.
Zuk, P. A., Zhu, M., Ashjian, P., De Ugarte, D. A., Huang, J. I., Mizuno, H., et al. (2002). Human adipose tissue is a source of multipotent stem cells. Molecular Biology of the Cell, 13, 4279–4295.
Uccelli, A., Pistoia, V., & Moretta, L. (2007). Mesenchymal stem cells: a new strategy for immunosuppression? Trends in Immunology, 28, 219–226.
Jiang, Y., Jahagirdar, B. N., Reinhardt, R. L., Schwartz, R. E., Keene, C. D., Ortiz-Gonzalez, X. R., et al. (2002). Pluripotency of mesenchymal stem cells derived from adult marrow. Nature, 418, 41–49.
Baksh, D., Song, L., & Tuan, R. S. (2004). Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy. Journal of Cellular and Molecular Medicine, 8, 301–316.
Musina, R. A., Bekchanova, E. S., Belyavskii, A. V., & Sukhikh, G. T. (2006). Differentiation potential of mesenchymal stem cells of different origin. Bulletin of Experimental Biology and Medicine, 141, 147–151.
Pasquinelli, G., Tazzari, P., Ricci, F., Vaselli, C., Buzzi, M., Conte, R., et al. (2007). Ultrastructural characteristics of human mesenchymal stromal (stem) cells derived from bone marrow and term placenta. Ultrastructural Pathology, 31, 23–31.
Reschiglian, P., Roda, B., Zattoni, A., & Bagnara G. P. (2007). Method and device to fractionate stem cell. Patent pending PCT/EP2007/054226.
Roda, B., Reschiglian, P., Zattoni, A., Alviano, F., Lanzoni, G., Costa, R., et al. (2009). A tag-less method of sorting stem cells from clinical specimens and separating mesenchymal from epithelial progenitor cells. Cytometry Part B (Clinical Cytometry), 76B, 285–290.
Biddison, W. E. (2001). Curr Protoc Cell Biol, Capter 2, Unit 2.2.