1. STEM CELL BIOENGINEERING Cláudia Lobato da Silva Research activities at Centro de Engenharia Biológica e Química

2. Large-scale ex-vivo expansion systems Control of self-renewal versus differentiation Stem Cell Bioengineering Determining the molecular and cellular mechanisms of stem cell expansion/differentiation and controlling culture conditions in an ex-vivo microenvironment – THE BIOREACTOR. STEM CELL BIOENGINEERING Stem Cell Bioengineering STEM CELL BIOENGINEERING

3. Stem Cell Expansion/Differentiation: whats the need? GOAL: increase the number of stem cells available (which is limited) while maintaining their in vivo engraftment potential and/or induce controlled cell differentiation into specific cell types. WHAT FOR? CELL THERAPY GENE THERAPY ALTERNATIVE TO ANIMAL MODELS STEM CELL EXPANSION / DIFFERENTIATION

4. Ex-vivo expansion of hematopoietic stem and progenitor cells from human bone marrow and umbilical cord blood: - co-culture with mesenchymal stem cell-derived stroma; - culture in stroma-free conditions. RESEARCH Main Research Topics Ex-vivo expansion and neural differentiation of mouse embryonic stem cells and mouse neural stem cells

5. HEMATOPOIETIC CELL EXPANSION Ex-vivo Expansion of Hematopoietic Cells in Serum-free Medium Lobato da Silva, C. et al, Experimental Hematology 33; 828-835 (2005)

6. Hematopoietic Stem Cell (HSC) Predictive Modelling Predictive Modeling self-renewal term k e X cell death term, k k X term accounting for the differentiation k d X limitation factor First-order kinetic including: Kinetic modeling can provide a significant insight into the "limiting steps" involved in the hematopoietic hierarchy and into the influence of conditions affecting the expansion process. Lobato da Silva, C. et al, Bioprocess Biosystems Eng., 25, 365-369 (2003) PREDICTIVE MODELLING

7. BM + hu-STBM - hu-ST CD34 + CD34 + CD38 - CD33 + Modelling Lobato da Silva, C. et al, Bioprocess Biosystems Eng., 25, 365-369 (2003) BM Modeling Results MODELLING

8. EMBRYONIC STEM CELL EXPANSION Fernandes, A.M. et al, Journal of Biotechnology (accepted for publication) (2007) Embryonic Stem Cell Expansion on microcarriers under stirred conditions

9. EMBRYONIC STEM CELL CHARACTERIZATION Fernandes, A.M. et al, Journal of Biotechnology (accepted for publication) (2007) Embryonic Stem Cell Expansion on microcarriers under stirred conditions MTT DAPI ALP Day 2 Day 4 Day 6 Day 8

10. ACKNOWLEDGEMENTS Acknowledgments Collaborations: University of Nevada, Reno, USA Rensselaer Polytechnic Institute, Albany, USA Institute of Molecular Medicine (IMM) Crioestaminal - Saúde e Tecnologia S.A. Financial Support: Fundação para a Ciência e Tecnologia, Ministério da Ciência Tecnologia e Ensino Superior