Application: Stem cells
Key objectives
Haematopoietic cells from peripheral blood or bone marrow - are effective targets for human gene therapy as they can be removed from the patient treated ex-vivo and then replaced. Stem cells would greatly increase the versatility of ex-vivo therapy techniques. This section sets out to evaluate the possible use of stem cells containing episomal gene expression vectors for application in gene therapy. The key objectives are to:
- Use pseudo-virus vector systems to optimize the efficiency of episomal DNA delivery to both dividing and non-dividing mammalian cells, including stem cells.
- Develop techniques for efficient expression from episomal gene delivery vectors in stem cells during long-term culture and define expression during stem cell self-renewal and commitment to differentiation.
- Use mouse embryonic stem cells to evaluate the behavior of episomal gene expression when these cells are induced to differentiate using cell culture or animal model systems.
Deliverables
Protocols will be defined for optimal delivery of epiosmal vectors into dividing and non-dividing cells using SV40 pseudo-virus particles. The pseudo-virus delivery protocols will be optimized for use with human and mouse stem cells. Multi-potent human stem cells will be used to evaluate the behavior of epiosmal gene delivery vectors during stem cell self-renewal and differentiation. Pluri-potent mouse embryonic stem cells will be used to evaluate the tissue specific expression of episomal gene delivery vectors in an animal model.