Post on 26-Mar-2015
ST
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STEM CELL BIOENGINEERING
Cláudia Lobato da Silva
Research activities at
Centro de Engenharia Biológica e Química
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.
ST
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Stem Cell Bioengineering
ST
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Stem Cell Expansion/Differentiation: what’s 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
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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.
RE
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Main Research Topics
Ex-vivo expansion and neural differentiation of mouse embryonic stem cells and mouse neural stem cells
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Ex-vivo Expansion of Hematopoietic Cells in Serum-free Medium
Lobato da Silva, C. et al, Experimental Hematology 33; 828-835 (2005)
PluripotentStem Cell
(PSC)
Stem Cell(SC)
MyeloidStem Cell
(MSC)
LymphoidStem Cell
(LSC)
CFU-G,M
CFU-MEG
BFU-E CFU-E
CFU-Eo
Granulocytes
Monocytes
Platelets
Erythrocytes
Eosinophils
T and B cells of theimmune system
Hematopoietic Stem Cell
(HSC)
Predictive Modelling
Pre
dict
ive
Mod
elin
g
Y
Xk
YdX
Xd
Xe XkXkPkXk
dt
Xd
• self-renewal term keX
• cell death term, kkX
• term accounting for the differentiation kd
X
• limitation factor
First-order kinetic including:
SCkSCkSCkHSCkSCkdt
SCdk
LySCd
MySCd
SCde
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)
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Experimental Theoretical
CD34+ CD34+CD38- CD33+
Modelling
Lobato da Silva, C. et al, Bioprocess Biosystems Eng., 25, 365-369 (2003)BM
Mod
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Fernandes, A.M. et al, Journal of Biotechnology (accepted for publication) (2007)
Embryonic Stem Cell Expansionon microcarriers under stirred conditions
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Fernandes, A.M. et al, Journal of Biotechnology (accepted for publication) (2007)
Embryonic Stem Cell Expansionon microcarriers under stirred conditions
MTT
DAPI
ALP
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AC
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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