ADSC-CARDIAC · Differentiation of adipose-derived stem cells info cardiomyocytes: Understanding and application
FP6 — Marie Curie Actions (Human Resources and Mobility)
- Duration
- 2004-11-01 → 2006-10-31
- EU contribution
- €80,000
- Participants
- 1
- Scheme
- IRG
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Results in brief
Final Activity Report Summary - ADSC-CARDIAC (Differentiation of adipose-derived stem cells info cardiomyocytes: understanding and application)
The projects main objective had been to investigate the potential of adipose tissue-derived stem cells to differentiate into cardiomyocytes and to search for specific inductors. Mesenchymal stem cells have been shown to differentiate into a variety of lineages such as bone, cartilage, fat, neurons and cardiomyocytes. The concept had been to establish immortalised clones that additionally would carry a cardiac-specific reporter construct, to screen in an automated fashion for factors that drive cardiomyocyte differentiation. Several research tools very developed during this project. Adipose derived stem cells were isolated from lipoaspirates from three donors and immortalised using retroviral constructs containing E6/E7 of the human Papiloma Virus. About 500 individual immortalised clones were isolated and tested for their differentiation capacity into cartilage, bone, fat and neuronal cells, following standard differentiation protocols. All clones showed a differentiation capacity albeit at different levels and different lineages. The concept of clonal cell differentiation capacity had been published for mesenchymal stem cells and was thus confirmed in this project for adipose derived stem cells. Mesenchymal stem cells are difficult to stably transfect, and hence a self-inactivating retrovirus was developed that allowed incorporation of cardiac-specific promoters coupled to a reporter gene (copGFP) and a selection cassette (neomycin). Several cardiac specific promoters were cloned and tested in embryonic stem cells for functionality. Embryonic stem cells are known for their high differentiation capability into cardiomyocytes. Upon transfer of these reporter constructs into the immortalised adipose-derived stem cell clones and treatment with the demethylating agent, 5-Azacytidine to induce cardiomyocyte differentiation, no reporter construct expression could be detected. Several groups had published the ability of mesenchymal stem cells to differentiate into cardiomycytes after 5-Azacytidine treatment. The lack of inducibility of the immortalised clones may be due to the immortalisation or lack of cardiomyocyte differentiation. Since all clones retained their differentiation capacity to other lineages, it is more likely that the adipose-derived stem cells are not suited to give rise to cardiomyocytes in a reproducible and possibly therapeutically interesting level.
Data: CORDIS, © European Union
Project objective
The main objective of the project described in this proposal is to provide a stable framework for the re-integration of a European researcher, who has been conducting academic research in the US for over 5 years, into a European SME. The project will allow the host institution to benefit from the knowledge and experience acquired by the applicant in his previous training and work in Europe and the US. The applicant's strong backgrounds in molecular and cellular biology, as well as developmental biology are as of significant interest to the company.As a result of this action, the applicant will continue his professional activity in his field of expertise in the European Union by conducting the company's stem cell differentiation project. Regarding the scientific content of this proposal, the applicant plans to utilize human adipose derived stroma (stem) cells to search for factors that initiate the differentiation into cardiomyocytes. The in vitro-differentiation of stem cells recapitulates, to our current under standing, the processes that take place during embryogenesis. Cardiac differentiation has been studied in great detail in vertebrates, and the expression of several genes during cardiogenesis delineates distinct developmental phases.We will use the promoters of these genes and couple them to fluorescent markers (GFP) and Luciferase, in order to achieve a qualitative and quantitative read out, respectively. One of the main objectives is to develop with the host SME a cell-based differentiation platform for high throughput screenings. Secondly, we will use a selection marker (puromycin) to obtain pure cardiac and pre-cardiac cell populations, which will be used for DNA-microchip analysis to search for new genes involved in cardiogenesis. The long-term goal i s to translate results from the chip analysis and high throughput screening into pre-clinical and clinical trials, using pre-differentiated (cardiac) cells for cell therapy purposes.
Original text from CORDIS.
Participants
- CELLERIX S.L. · TRES CANTONS (MADRID)CoordinatorCountry levelSpain
Links
Data: CORDIS, © European Union