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Mesenchymal Stem/Progenitor cells
for Therapeutic and Tissue
engineering applications
Yu-Hui Tsai, Ph.D.
( Daniel Tzu-bi Shih)
Grad. Inst. Medical Sciences
Taipei Medical University
Stem/Progenitor Cells
Stem cells
• Self Renew (Proliferation)
• Muti-differentiation potential (Plasticity) :
Trans-differentiation, Transformation, Dedifferentiation
Progenitor cells
• Functional Undetermined Tissue-cell
Precusors.
• Muti-differentiation potential (Plasticity)
Types of Stem Cells
– ES Cells (ESCs) [Derived from Embryo inner
mass]
– EG Cells (EGCs) [Derived from Fetus gonad]
– Somatic Tissue Stem /Progenitor Cells
[ Isolated from Fetus, Neo-natal and Adult
Tissues] (tSC, tS/PC): e.g. Neural SCs,
Mesenchymal Stem/Progenitor Cells (MSC,
MSCs/MPCs), Hemapoietic Stem/Progenitor
Cells (HSC, HSCs/HPCs), and Karotinocytes
etc.
Development of the Human Preimplantation Blastocyst
Development of Human Embryonic Tissues
The Niche
Determines The Fate
of Stem Cells
Ecologic/Epigenetical
Microenvironment of
Stem Cells
Tissue Stem Cells (tSCs)
Develop & Maintain within Specific Niche
A. Epidermal---Basal Layer
Hair Folocale & Epidermis--- The bulge (Tcf3)
B. Small Intestine--- The crypt Cells ( Tcf4)
C. Neural Precursors Cells--- Ependymal or the Sub
Ventricular Zone Cells (AP)
D. Hematopoietic SCs--- Bone Marrow
Tissue Maintenance and Repair
• I) Tissue Stem/Progenitor Cells
• II) Environmental Cellular Metrix (ECM)
• III) Associate Stroma Cells
• IV) Cytokines
Basic Requirements for Tissue
Regeneration
*Tissue Stem/Progenitor Cells
*Stromal Cells for MicroEnvironmental Support of the
Tissue Survival and Normal Functioning.
*Cytokines (Autokine & Parakines) & Chemokines
*Oxygen Supply:
Red Blood Cells and Neovascularization
*Sensory Nuron: nurogenesis.
Wo u n d
H e a l i n g
Adapted from speech of Dr. Bruce A. Mast, M.D.: The Regulation of Wound Healing.
Division of Plastic and Reconstructive Surgery Department of Surgery Univeristy of
Florida. (http://www.medinfo.ufl.edu/cme/grounds/mast/index.html).
CD34+ HS/PCs Functions:
Hematopoietic Reconstitution
Immunotherapy
Neovasculogenesis, Neurogenesis,
&Tissue Regenerations etc.
Comparative analysis of the similar stem cell
populations from various origins
The Niche of HSCs
Marrow
(MSC & ECM)
& Soluble Factors
Tissue Mesenchymal Stem Cells
Definition:
▪Cell surface antigens expression : SH2/SH4/CD105
▪Mesenchymal Multi-lineage differentiation potential
Sites :
▪
Connective tissue / stromal source
EX : Bone marrow /Adipose tissue /
Skin / Muscle
▪
Umbilical cord blood
Bone Marrow Mesenchymal stem
cells ( BM-MSCs )
Multipotent differentiation
(mesoderm-derived tissue cell type) : adipocyte /
osteoblast / chondrocyte / myoblast / endothelial
cell
(Ectoderm) : Neural cells
Interact & Support tissue cells
Hematopoiesis and angiogenesis
Hypotheses to explain the generation of nonhematopoietic tissues from HSC-enriched populations. Totipotential cells derived
from the embryo could persist into adult life and be sequestered in the bone marrow, where they could form a reservoir of
precursors from which definitive HSCs could develop. Protocols for the enrichment of HSCs based on their phenotypic and
physical properties might also result in the isolation of these totipotent cells, which in turn contribute to skeletal or cardiac
muscle or cells of the gastrointestinal tract after various transplantation protocols. Alternatively, precursors whose
developmental potential is more limited might also be present in the bone marrow. Finally, a pluripotent HSC (PHSC) may be
Stem cells find their niche
Creation of long-lasting blood vessels
NATURE|VOL 428 |11 MARCH 2004|
Gene expression profiling of BM-MSCs
Multipotent :
Undifferentiated human mesenchymal stem cells reveals
mRNAs of multiple cell lineages.
Tremain,N.2001 / Woodbury,D.2002 / eshi,B.2003
Growth factors / Matrix proteins / Cell surface
molecules :
BM-MSCs expressed various kinds of growth factors,
matrix proteins, and cell surface molecules.
Silva,W.A.,Jr2003
Adipose tissue Mesenchymal stem
cells (AD-MSCs)
• Similar to Bone marrow mesenchymal stem
cells
▪ CD marker antigens
▪ Multipotent differentiation
A Study on Human Tissue Stem/progenitor cells for
Medical Application and Drug Discovery
• Bone Marrow
• Placental and
Peripheral Blood
• Fat Tissue
• Scalp
• Amnoitic tissue
• Fore Skin
• Placenta
• Teeth
Stem Cell Isolation and Processing
a. Hematopoietic Stem Cells (HSCs) and Mesodermal Progenitor
Cells (MPCs)
MACS
CD45-GlyA- cells (MPCs)
MNC
FACS
CD34+ cells (HSCs)
c. Processed Lipoaspirate (PLA)
Adherent cells -PLA
How to distinguish tissue stem cells ?
Cell surface antigens expression
Multi-lineage differentiation abilities
Is There Specific Niche for Each Type of
Tissue Stem/Progenitor Cells in Human?
Objective : To understand the tissue MSC
specificity for better stem cell therapy applications
Hypothesis: Mesenchymal stem cells in
different tissues have their own specific potentials
in maintenance of their tissue specific niche
How Specific Are Tissue Niche Need?
By Combining Cellular and Molecular
Characterizations of MSCs derived from
Different Tissues in Human, ….
Strategy :
Comparative analysis gene profiling and
functional gene characterization
Target gene expression
Multi-lineage differentiation related genes
Early gene expressions
Highly differential expression genes
Cytokines and growth factors genes
Confirm the protein products
Functional genomic characterization
Part 1. Multi-lineage differentiation related genes
ostogenic /
chondrogenic /
adipogenic /
myogenic /
neurongenic /
endothelial cell differentiation
Molecular Analytic results suggest that BMMSCs preferentially expressed functional
genes differences from AD-MSCs in:
•Secretion of Hemagiogenic factors
(LIF / Jagged-1/ BMP-4 / HGF / PIGF-1 / PIGF-2
/ HB-EGF ),
•Supply Homing/ mobilzing elements
(:CD106(VCAM-1)/ jagged-1/ SDF-1), and
•Generation of ECM remodeling factors
(mmp9/mmp1) for HS/PCs (CD34+, CD38-, and
CXCR-4+)
Highly differential expression of genes is
primarily involved in
Growth factors /
Extra cellular matrix molecules /
Cell surface molecules /
Receptors /
▪
Suggest that the major difference
between BM-MSCs and AD-MSCs :
cell-cell interactions
regulate or support tissue cells
1. The tissue cells homing and mobilization.
CD106 (VCAM-1) /
SDF-1 /
MMP-9/
HGF
Journal of Cellular biochem S 38:29-38 (2002)
BM-MSCs exppressed more key factors involved in
Homing and mobilization of hematopoietic stem cells :
CD106 / SDF-1 / MMP-9
2. The tissue cells supporting.
Hematopoiesis
LIF / SDF-1 / BMP-4 / Jagged-1 / HGF
Angiogenesis
HGF / PIGF-1 / PIGF-2 / HB-EGF
3. The ECM remodeling properties
ECM remodeling protease
MMP-1 / MMP-9
Conclusions
Our present study indicated the tissue specificity
of mesenchymal stem cells exhists as analyzed
by molecular characterization of their gene and
protein expressions.
In addition to their cell surface phenotypic
descriptions and multipotency examinations in
vitro, this approach may be indispensable in
further realizing the nature of tissue stem cell
populations
Thanks for
Your Attensions
The Haematopoietic System
• Humoral Regulation
Multipotent
Stem Cell
Modulating
IL1
IL4
MIP-1
TNF
TGF
Self-renewal
?
CMPs
CLPs
CFU-G, E, M, Mega
BFU-E
SCF
IL3
GM-CSF
CFU-E
SCF
IL3
IL6
Tpo
SCF
IL3
GM-CSF
CFU-G
SCF
IL3
IL6
G-CSF
GM-CSF
Epo
IGF-1
Proerythroblast
CFU-GM
CFU-Mega
Megakaryocyte
GM-CSF
M-CSF
Myeloblast
G-CSF
GM-CSF
Epo
IGF-1
CFU-M
Monoblast
GM-CSF
M-CSF
CFU-Eo
GM-CSF
IL5
Eosinophilic
Monoblast
IL5
CFU-Baso
IL3
IL4
Basophilic
Monoblast
IL3
IL4
Tpo
Erythrocyte
Platelet
Neutrophil
Monocyte
GM-CSF
M-CSF
Macrophage
Eosinophil
Basophil
IL3
IL4
Mast Cell
Early Acting
SCF
FL
Tpo
G-CSF
IL3
IL6
IL7
IL11