Stem Cells and

Regenerative Medicine
Andi Wijaya, 2012.-

Life is
Regenerative

Life is regenerative, by definition.-

Regenerative medicine seeks to devise new

therapies for pts with severe injuries or chronic
diseases in which the bodys own responses do
not suffice to restore functional tissue.-

Atala et al, Foundation of Regen Med 2010.2

Salamander :
Natures Tissue Engineer
MSCs in the Blastema
3

Sir Martin Evans

Nobel Prize 2007
For Generation of
Mouse ESCs.4

Dr. James Thomson

Created first hESCs
In 1998.-

Prof. Sinya Yamanaka

Created iPS cells
from somatic cells
in 2006.-

Generation of iPS cells

JCI 2010.-

Sir Ian Wilmut

The Cloning of Dolly
8

Cloning by somatic cell nuclear transfer

Foundation of Regen Med, 2010.-

Prof. Anthony Hollander

Tissue Engineering
Pioneer

10

Chondrocyte transplantation in the right femoral

condyle

Foundation of Regen Med, 2010.-

11

What Is
Stem Cell ??

Stem Cells are defined as self-renewing

cell populations that can differentiate
into multiple distinct cell types.-

Embryonic stem cells which are capable

of virtually unlimited proliferation &
differentiation (pluripotent).-

Adult stem cells, which can generate a

for more limited repertoire of
differentiated cell types.12

Stem cells
sperm

Adult stem cells

Neurones

Fetilised eeg Totipotent cells blastocystEmbryonic stem cells

Bone tissue

eeg

Fertilisation Stimulated
to develop

Blood cells

Five days
growth

Differentiation into Specialisation into

final cell types
adult stem cells

13

Cell lineages from the ICM of human

blastocysts
ICM
Human blastocyst
TE

ICM (hES cell)

Epiblast

Hypoblast
Yolk sac

Ectoderm

Skin

NS

CT

Muscle

Cardiac

Stem Cells, 2005.-

Skeletal

Mesoderm

Eyes

UGS

Ears RS

VS

Smooth

Bone

Endoderm

GI

Germ cells

Liver PancreasSperm

Cartilage

BM

Eggs

Embryonic Stem
(ES) Cells

ES cells are derived from the inner cell

mass (ICM) of blastocysts (embryos that are
~ 3,5 days post coitum).-

The ICM of blastocysts contains all of the

cells that will give rise to the embryo it self
& the primitive endoderm.-

In the embryo, ICM cells partially

proliferate, but all of the differentiate &
lose their pluripotency within a short period
of time.-

Nishikawa et al, Nat Rev Mol Cell Biol 2007.-

Generation of embryonic stem cells from blastocyst-stage

embryos

Foundation of Regen Med, 2010.-

16

Adult
Stem Cells
Bone Marrow-Derived SCs:
1. Hematopoietic SCs (HSC).2. Progenitor Cells (Precursor Cells).3. Mesenchymal SCs (MSC). Organ Specific SCs ( NSC, CSC etc). Induced Pluripotent SCs (iPSCs).17

Fetal Stem Cells

(Fetal MSCs) (1)
The recent isolation of fetal SCs from several
sources either at the early stages of development
or during the later trimester of gestation, sharing
similar growth kinetics & expressing
pluripotency. These provide strong support that these cells may
be biologically closer to ESCs.18

Poppa & Anagnou, regen Med 2009.-

Fetal Stem Cells

(Fetal MSCs) (2)
Fetal SCs actually representing intermediates
between ESCs & adult MSCs, regarding
proliferation rates & plasticity feature. Thus able to confer an advantage over postnatal
MSCs derived from conventional adult source
such as BM.19

Poppa & Anagnou, Regen Med 2009.-

Fetal Stem Cells

(Fetal MSCs) (3)
Fetal SCs represent a relative newcomer in the
filed, exhibiting unique & fascinating features. These cells can be derived either from the fetus
proper or from the supportive extra embryonic
structures that are of fetal origin, incl. UCB,
amniotic fluid (AF), amniotic membrane,
Whartons jelly, & the placenta.20

Poppa & Anagnou, Regen Med 2009.-

Basic steps of fetal formation and maturation during human

gestation at the level of tissue structure, depicting the
major fetal sources harboring stem-cell populations

Regen. Med, 2009.-

21

Current model of the developmental position and potency of

the fetal stem cells from several fetal sources emerging
during gestation, such as the aminotic fluid, Whartons jelly,
placenta and aminotic membrane

Regen. Med, 2009.-

22

Umbilical Cord Blood

Stem Cells (UCB)

UCB contains both HSC & MSC.-

Because stem cells in UCB exist in

higher numbers than in adult human
blood or BM.-

A population of cells from human UCB

called unrestricted somatic stem cells.23

Regenerative
Medicine

Regenerative medicine aims to repair

diseased or damaged tissues by
replacing the affected cells with
healthy, functional cells of the same
type.-

The prospects of this discipline have

been boosted by the promise of ES
cells, which are pluripotent (they can
differentiate into any cell type) & which
can be maintained in culture to selfrenewal indefinitely.-

Blelloch, Nature 2008.-

Bone Marrow
Stem Cells (BMSCs)
The most well characterized source for adult SCs
is the adult BM. Adult BM contains a heterogeneous population of
cells, incl. HSCs, macrophage, fibroblasts,
adipocytes & EPCs. BM also contains a subset of non-HSCs that
posses a multilineage potential, these SCs are
called marrow stromal cell, or mesenchymal SCs,
& now term mesenchymal stromal cells (MSCs).Xaymardan et al, SC Anthrology 2008.-

25

Mesenchymal
Stem Cells (MSCs)
Mesenchymal Stem Cells (MSCs) also known as
multipotent mesenchymal stromal cells are selfrenewing cells that can be found in almost all
postnatal organs & tissues. The ability of MSCs to differentiate into several
cell types incl. muscle, brain, heart, skin, fat,
bone & cartilage cells make them attractive as
therapeutic agents for a number of diseases.26

Volarevic et al, Stem Cells, 2011.-

MSCs as Potential
Cellular Therapy
Their ease of isolation from BM, adipose tissue,
Whartons jelly, UCB & placenta, high ex-vivo
expansion potential & ability to differentiate into
multiple lineages, make them attractive tools for
potential use in cell therapy. The immunosuppressive properties of hMSCs
make them an important candidate for cellular
therapy in allogenic setting.27

Kode et al, Cytotherapy 2009.-

Schematic representation of mesenchymal stem cell

development pathways

Foundation of Regen Med, 2010.-

28

Pluripotency of BMSCs

Foundation of Regen Med, 2010.-

29

Immunodulatory
Properties of MSCs
MSCs have been reported to be
immunoprivileged cells, a relevant feature since it
would allow their allogenic or xenogenic
transplantation. Although the mechanisms underlying the
immunosuppressive effects of MSCs have not
been clearly defined, it seem that MSCs
modulate the function of different cells involved
in the immune response.30

Gracia Gomez, Expert Opin Biol Ther 2010.-

Immunomodulation
Characteristics of MSCs (1)
The immune phenotype of MSCs (widely
described as MHCI+, MHCII-, CD40-, CD80-,
CD86-) is regarded as non-immunogenic and,
therefore transplantation into an allogenic host
may not require immuno suppression.31

Chamberlain et al, Stem Cells 2007.-

Immunomodulation
Characteristics of MSCs (2)
MSCs also having immunosuppresive properties
specifically MSCs can modulate many T-cell
functions including cell activation. MSCs also have immunodularly properties
impairing maturation & function of dendritic
cells & MSCs inhibit human -cell proliferation,
differentiation & chemotaxis.32

Ghannam et al, SC Res & Therapy 2010.-

Simplified scheme of the MSCs effects on cells of immune

system

Exper Opin Biol Ther, 2010.-

33

MSCs Based
Therapeutic Activities

Injected or infused MSCs based on 2 generalizable

therapeutic activities :
Immunomodulation has been shown to be mediated
by both secreted bioactive molecules & by cell cell
contact, can involve dendritic cells & T & B cells,
incl. T-reg, killer cells & T-helper cells.Trophic effects, involved MSC-secreted molecules
that inhibit apoptosis (especially caused by
ischemia) & scar formation, also involved
stimulation of MSC-mediated angiogenesis, (VEGF)
34
& stabilization of new vessels (pericyte).-

Caplan & Correa, Cell Stem Cells 2011.-

Companies Developing
MSC Therapies (1)
1.

Osiris Therapeutic Inc (USA)

Prochymal : for GvHD. Provacel : for AMI. Chondrogen : for knee injury.-

2.

Mesoblast (Australia)

MPCs (mesenchymal precursor cells) :

for Osteoarthritis & bone repair, for Heart failure &
PAD.3. Brainstorm Cell Therapeutic Inc
NurOwn : for Neurodegenerative diseases.4. Athersys (USA)
Multistem : for IBD, AMI, GvHD.Giordano et al, J Cell Physiol 2007.-

35

ES & iPS Cells

In Regenerative Medicine

Although ES cells are promising donor sources

in cell transplantation therapies, they face
immune rejection after transplantation & there
are ethical issues regarding the usage of human
embryos.-

These concerns may be overcome by using iPS

cells, that can be directly derived from pts
somatic cells.36

Yamanaka et al, Nature 2007

Four experimental routes for nuclear reprogramming

A.
Nuclear
transfer
to eggs

C.
Lineage switch

D.
Direct
conversion

B.
Pluripotency
(iPS)

Adult
cell
types

Sience 2008.-

37

Examples of transcription factor overexpression or

ablation experiments that result in cell fate changes

Science 2008.-

38

Hypothetical strategy for using iPS cells

in cell-based therapies

Nature Med 2007.-

Cord Blood
Banks

Cord blood is usually stored for future

allogenic use.-

To date, approximately 1,3 million cord blood

units have been stored worldwide, while some
15 of those have been used for transplantation
so far.-

Pamphlets & web pages of private cord blood

banks invariably contain some reference to the
future use of cord blood cells for treatment of
heart disease, a concept that is likely to
influence the decision making process of
future parents.-

Stamm & Nan Ma, SC Repair & Regen 2008.-

Suggested concept for the clinical use

of autologous cord cells in children
Diagnostic of
congenital
defects
Harvesting of
umbilical
cord cells

Isolation of
Vascular cells
Implantation
into the same
individiuum
at an appropriate
time for sugery

Storing cells
in cellbanks

Tissue engineering
of heartvalves
(or cardiovasc.
constructs)

Stem Cell Repair & Regeneration, 2008.-

Reculturing of
Thawed cells

N2

Stromal Vascular Fraction (SVF)

& Adipose Derived Stromal /SCs (ASCs)
Subcutaneous fat is an abundant & accessible
source of both uncultured/heterogeneous SVF
cells & cultured / relatively homogeneous ASCs, The peer-reviewed literature focusing on SVF
cells & ASCs research has expanded
exponentially over the past decade, And demonstrated by the registration of 36
clinical trials in 11 different countries,42

Gimble et al, Stem Cells 2011,-

The process of ASC clinical usage

Stem Cells 2011.-

SCs for Regenerative

Medicine Applications
Ideally, SCs for regenerative clinical applications
should meet the following criteria :
1. Can be found in abundant quantities (106-109),2. Can be harvested by minimally invasive procedure,3. Can be differentiated along multiple cell lineage in
regulatable & reproducible manner,4. Can be safely & effectively transplanted either
autologous on allogeneic,5. Can be manufactured with GMP guideline,Ginible et al, Circ Res 2007,-

44

Safety issue in
ES Cell Therapy

In view of the intrinsic ability of ES

cells to form teratoma (a type of tumor)
the major safety issue of FDA was how
to ensure the nontumorigenity of the
cells used for theraphy.-

The purity of the differentiated cells is

thus a safety concern.-

Editorial Nat Med 2008.-

Stem Cell-Based
Therapies

Recent record the listing of 69

different human illnesses being
treated by adult & fetal stem cells.-

Legislation still depends on

persuasion, on bargaining, on the
political process.-

If science & especially technology

move like lightning, the image for
legislation approximates molasses.46

Acknowledgments

47