The twitcher mouse is an animal model of Krabbe's disease which is a neurodegenerative lysosomal storage disorder resulting from the absence of functional lysosomal enzyme galactocerebrosidase (GALC). This disease affects the central and peripheral nervous systems and in its most severe form results in death before the age of 2 years old in humans and approximately 30-40 days in the mouse.
Adipose derived- and bone marrow-derived murine mesenchymal stem cells (mMSCs) may be used to study stem cell properties in an in vivo setting for the purposes of evaluating therapeutic strategies that may have clinical applications in the future. One solution to the problem of how to track the administered cells is to transplant cells with an easily identifiable genetic marker such as enhanced green fluorescent protein (eGFP). The expression of eGFP does not appear to affect the ability to differentiate along adipogenic or osteogenic lineages; however it appears that the tissue of origin can influence differentiation capabilities. The presence of eGFP had no effect on cell surface marker expression, and there were no significant differences between transgenic and non-transgenic mMSCs.
Subsequent to the intracerebroventricular injection of eGFP+ transgenic ASCs or BMSCs on post-natal day (PND) 3-4, body weights, lifespan, and neuromotor function were evaluated beginning on PND 15. Tissues were harvested for analysis of GALC activity, presence of myelin, infiltration of macrophages, microglial activation, inflammatory markers, and cellular persistence.
Survival analysis curves indicate a statistically significant difference in lifespan between stem cell treated and control twitcher mice. Body weights and motor function were also improved. The stem cells may mediate some of these benefits through an anti-inflammatory mechanism because the expression of numerous pro-inflammatory markers were down-regulated at both transcriptional and translational levels. A marked decrease in the levels of macrophage infiltration and microglial activation were also noted.
The data from these studies indicate eGFP+ mesenchymal lineage stem cells can be tracked post-injection, are potent inhibitors of inflammation associated with Krabbe's disease progression, and reduction in the levels of inflammation will be beneficial as a component of a combination approach for in vivo treatment.
|Advisor:||Bunnell, Bruce A.|
|Commitee:||Abdel-Mageed, Asim, Beckman, Barbara, Betancourt, Aline, Mondal, Debasis|
|School Location:||United States -- Louisiana|
|Source:||DAI-B 72/02, Dissertation Abstracts International|
|Subjects:||Cellular biology, Pharmacology|
|Keywords:||Adults, Globoid cell leukodystrophy, Krabbe's disease, Mesenchymal stem cell, Regenerative medicine, Stem cells|
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