Key to advancing stem cell utilization in regenerative medicine and cell-based therapies is the development of systems to expand cells to clinically relevant numbers while maintaining the desired stem cell phenotype. Mathematical growth models play an important role in developing standardized systems, as they are both predictive tools for expansion potential and tools to describe current kinetic parameters of a stem cell population. One disease that may benefit from cell therapy is Duchenne Muscular Dystrophy (DMD), a muscle disease characterized by the lack of dystrophin expression at the sarcolemma of muscle fibers resulting in muscle fiber necrosis and muscle weakness. While transplantation of normal myoblasts into dystrophin-deficient muscle can restore dystrophin, the use of muscle-derived stem cells (MDSC) has enhanced the success of cell transplantation. For these reasons, muscle stem cell isolation and the development of transplantation techniques have garnered increased attention recently. One limitation of MDSC use is the few numbers of cells available from a muscle biopsy, thus presenting the requirement for in vitro expansion. The overall goal of this study was to provide a thorough quantitative examination of the expansion of MDSC populations. In this project, an imaging system was established to analyze stem cell expansion. The applicability of this system was demonstrated in MDSC expansion with cytokine stimulation. It was found that accounting for the proliferative heterogeneity that exists in stem cell populations would allow for more accurate estimations of kinetic parameters. Next, a more sophisticated imaging system was used to further develop an automated system for analysis of MDSC proliferation and behavioral characterization. Finally, an understanding of the limits of expansion was explored. The role of long-term expansion on stem cell phenotype and regeneration capacity was examined to consider the issue of quantity vs. quality of muscle-derived stem cells. This study provided a systematic method for assessing expansion and an in-depth investigation into the natural progression of stem cell expansion. It is expected that these findings will provide a biological understanding of the limits of expansion and a foundation for more standardized methods of expansion of MDSC as MDSC are advanced to a clinical setting.
| Identifer | oai:union.ndltd.org:PITT/oai:PITTETD:etd-06292004-110834 |
| Date | 13 September 2004 |
| Creators | Deasy, Bridget M |
| Contributors | Bard Ermentrout, Ph.D, Johnny Huard, Ph.D., William Wagner, Ph.D, Bruno Peault, Ph.D |
| Publisher | University of Pittsburgh |
| Source Sets | University of Pittsburgh |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.pitt.edu/ETD/available/etd-06292004-110834/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Pittsburgh or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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