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Dissertation
Murine bone marrow stromal cells: isolation, characterization, and differentiation
Doctor of Philosophy (Ph.D.), Medical College of Pennsylvania and Hahnemann University
Dec 1999
DOI:
https://doi.org/10.17918/00007738
Abstract
In adult bone marrow, hematopoiesis is modulated by both direct, cell-cell contact with stromal cells, and by the chemokines and extracellular matrix that they elaborate. In addition to their role in supporting hematopoiesis, bone marrow stromal cells have also been shown to represent a unique population of pluripotent progenitors of non-hematopoietic connective tissues. Friedenstein et al. (1966&1970) first showed that stromal cells could be separated from blood-forming cells by their propensity to adhere to tissue culture plastic and then differentiated into cells of the osteogenic lineage. He later showed (Friedenstein et al. 1987) that this osteogenic precursor derived from the fibroblast-like marrow stromal cell (MSC). Since then, many others have extended Friedenstein's original observations to show that MSCs can also be differentiated into cartilage, fat, and muscle. Because of the plasticity of these cells and the ease by which they can be isolated form posterior iliac crest and expanded ex vivo, MSCs have become an attractive vehicle for cell and gene therapy. Our lab has spent the last several years characterizing the growth and differentiation potential of both murine and human MSCs. We have found however, in agreement with others, that while plastic adherent, human bone marrow cultures result in a relatively pure fraction of MSCs after several passages (Phinney et al. 1999b), murine explants yield cultures that are complex in composition and often difficult to enrich for MSCs by serial passage alone. We have also shown that the yield and growth kinetics of MSCs in standard liquid cultures vary significantly even between different inbred stains of mice. These limitations have, therefore, hampered progress in developing murine models by which to study both the in vitro and in vivo potential of these cells. As a result, we have devised a simple isolation procedure for generating a highly enriched fraction of murine MSCs that can be differentiated in vitro into osteocytes, chondrocytes, and adipocytes, and in vivo into osteocytes and astrocytes.
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Details
- Title
- Murine bone marrow stromal cells
- Creators
- Gene C. Kopen
- Contributors
- Darwin J. Prockop (Advisor) - Drexel University, Medical College of Pennsylvania and Hahnemann University (1993-1996, 1998-2002)
- Awarding Institution
- Medical College of Pennsylvania and Hahnemann University
- Degree Awarded
- Doctor of Philosophy (Ph.D.)
- Publisher
- Medical College of Pennsylvania and Hahnemann University; Philadelphia, Pennsylvania
- Number of pages
- x, 151 pages
- Resource Type
- Dissertation
- Language
- English
- Academic Unit
- School of Medicine (1993-1996, 1998-2002); Medical College of Pennsylvania and Hahnemann University (1993-1996, 1998-2002)
- Other Identifier
- 991021889012504721