CERTIFICATE INTRODUCTION TO REGENERATIVE MEDICINE:

BIOLOGY AND APPLICATIONS OF HUMAN STEM CELLS

6ª Edición (2017-18)

STEM CELLS:
Introduction to Biology and Technology

Deborah Burks
Centro de Investigación Principe Felipe
 Cell Therapy
• “autologous, allogeneic, or xenogeneic cells that
have been propagated, expanded, selected,
pharmacologically treated, or otherwise altered in
biological characteristics ex vivo to be administered
to humans and applicable to the prevention,
treatment, cure, diagnosis or mitigation of disease or
injuries”
• October 14, 1993. 58 FR 53248
Advantages of Cell therapy
• Cells are dynamic. They migrate, proliferate,
differentiate, and respond to their
environment in vitro and in vivo
• Multiple cell types and mechanisms of action
can be involved
• Therapeutic outcome can be curative and
permanent
– Repair, replace, regenerate
 Stem Cell History
1998 - Researchers first extract stem cells from human
embryos. Thompson et al, H9 and H1.
1999 - First Successful human transplant of insulin-making
cells from cadavers. Edmonton trial.
2001 - President Bush restricts federal funding for
embryonic stem-cell research
2002 - Juvenile Diabetes Research Foundation
International creates $20 million fund-raising effort
to support stem-cell research
2002 - California ok stem cell research
2004 - Harvard researchers grow stem cells from embryos
using private funding
2007 - Introduction of cellular reprograming to generate
human induced pluripotent cells (iPS)
2012 - John B. Gurdon and Shinya Yamanaka awarded
Nobel prize in Medicine for iPS technology
 Stem Cell – Definition
• A cell that has the ability to continuously
divide and differentiate (develop) into
various other kind(s) of cells/tissues
Why are stem cells special?
Stem cells can:
• self-renew to make
more stem cells
• differentiate into a
specialised cell type

Stem cells that can become Stem cells that can become
many types of cells in the only a few types of cells are
body are called pluripotent called multipotent

Embryonic stem cells (pluripotent) Tissue stem cells (multipotent)

 Degrees of Stemness
Stem cell
type Description Examples
Cells from early
Each cell can develop
Totipotent (1-3 days)
into a new individual
embryos
Some cells of
Cells can form any (over
Pluripotent blastocyst (5 to 14
200) cell types
days)
Cells differentiated, but Fetal tissue, cord
Multipotent can form a number of blood, and adult
other tissues stem cells
 Types of Stem Cells

Embryonic stem cells come from a five to six-day-old embryo. They

have the ability to form virtually any type of cell found in the human
body.

Embryonic germ cells are derived from the part of a human embryo or
foetus that will ultimately produce eggs or sperm (gametes).

Adult stem cells are undifferentiated cells found among specialised or

differentiated cells in a tissue or organ after birth. Based on current
research they appear to have a more restricted ability to produce
different cell types and to self-renew.
Tissue stem cells
• often known as adult stem cells
• also includes stem cells isolated from fetal and cord
blood
• reside in most tissues of the body where they are
involved in repair and replacement

Bone marrow Kidney Lung

• generally very difficult to isolate

• already used to treat patients (haematological
malignancies, diseases of the immune system)
Where do embryonic stem cells
come from?
• Donated excess IVF embryos Inner cell mass

egg fertilised 2-cell 8-cell blastocyst

egg

Day 0 Day 1 Day 2 Day 3 Day 6

 Stages of Embryogenesis

Day 2
2-cell embryo Day 3-4
Day 1
Multi-cell embryo
Fertilized egg

Day 5-6
Day 11-14 Blastocyst
Tissue Differentiation
Blastocyst Diagram
Derivation and Use of Embryonic Stem
Cell Lines
Isolate inner cell mass
Outer cells (destroys embryo)
(forms placenta)
Inner cells
(forms fetus) Culture cells

Day 5-6
Blastocyst “Special sauce”
(largely unknown)

Heart
Dr. Hariom Yadav
repaired
Kidney Heart muscle
Dr. Hariom Yadav
 Embryonic stem cells: Issues
• Pros:
– Pluripotent cells
– Can be induced to differentiate into basically all adult cell types

• Cons:
– Ethical/Religious issues concerning use of human ES cells
– Can form teratomas. Safety?
• therefore generally used to generate more differentiated cell types like
Neuronal SCs, Muscle progenitor cells, MSCs etc.
– Immunological problems - foreign cells
• need to use immuno suppression
• Nuclear transfer - cloning: Somatic nuclear transfer into ES - dolly the
sheep. Epigenetic issues
– HuESCs Grown on a feeder layer of mouse embryonic fibroblasts (MEFs) and
require the presence of basic Fibroblast Growth Factor (bFGF or FGF-2). Work
on developing defined culture conditions without animal products - needed for
clinical use.
• Treatments:
– There are no approved treatments using embryonic stem cells. Geron, a
pharmaceutical company, received clearance from the FDA in January 2009 to
begin the first human clinical trial of an embryonic stem-cell based therapy in
the treatment of acute spinal cord injury. Used oligodendrocyte progenitor cells
(OPCs) that have been derived from human embryonic stem cells. the clinical
Low efficiency of Differentiation

D’amour et al
WHAT ARE INDUCED PLURIPOTENT STEM CELLS?

Induced pluripotent stem cells (iPS cells) are a type of pluripotent

stem cell artificially derived from a non-pluripotent cell, typically an REPROGRAMMING VIEW
adult somatic cell, by inducing a "forced" expression of certain genes.

Warren et al., 2010. Cell Stem Cell

USES OF iPS CELLS – Gene and/or Cell therapy
Embryonic vs Adult Stem Cells
 Stem cell function in injury response

•Factors Critical to repair: Getting the cells to where they

are needed
– Stem cell mobilization
•G-CSF/SDF-1 regulation of mobilization

– Stem Cell Homing

•Chemotaxtic gradients

– Stem cell activity in injury site

•Migration at site
•Paracrine support
•Cell differentiation
 Stem Cell Niches
• There is strong evidence that the behavior of stem cells is strongly affected by their
local environment or niche.

• Some aspects of the stem cell environment that are known to influence self-renewal
and stem cell fate are:
– adhesion to extracellular matrix proteins
– direct contact with neighboring cells
– exposure to secreted factors
– physical factors, such as oxygen tension and sheer stress

• Manipulation of these factors underlies Tissue Engineering

•and maybe critical in maintaining stem cell populations during aging

 Stem Cell Niche

- Niche usually associated with blood vessel

- Frequently low O2 tension

1) Divide into 2 stem cells

2) Divide into 2 differentiated cells
3) Asymmetric division (stem & Differentiated cells)
4) Diff - dediff into stem cell
Adult neural stem cell populations
Ischemic damage induces neurogenesis

Triple BrdU NeuN DNA

Bone Marrow Stem Cells
 Adult Stem cell-based therapies are
already in clinical use
• Hematopoietic stem cell transplantation is the oldest (40 + years)
stem cell therapy and is the treatment that is most widely available.

• The stem cells come from bone marrow, peripheral blood or cord
blood. For some applications, the patient's own cells are engrafted.

• Allogeneic stem cell transplantation is now a common procedure for

the treatment of bone marrow failure and hematological
malignancies, such as leukemia.

• Donor stem cells are used to reconstitute immune function in such

patients following radiation and/or chemotherapy.
 Applications
 Disease
• Diabetes, Spinal
cord injury,
Parkinson’s
disease, heart
disease
 Genetic based
Disease
• Cystic fibrosis,
Huntington’s
10/20/2016 Dr. Hariom Yadav
 Stem Cells are Tool:
Regenerative Medicine
• Rejuvenation. Rejuvenation means boosting the body's natural
ability to heal itself. Though after a cut your skin heals within a few
days, other organs don't repair themselves as readily.

• Replacement. Replacement involves using healthy cells, tissues

or organs from a living or deceased donor to replace damaged ones.
Organ transplants, such as heart and liver transplants, are good
examples.

• Regeneration. Regeneration involves delivering specific types of

cells or cell products to diseased tissues or organs, where they will
ultimately restore tissue and organ function. This can be done
through cell-based therapy or by using cell products, such as growth
factors. Bone marrow transplants are an example.
 Cell Therapy and Tissue Engineering

Autologous vs Allogenic?
How to “home” cells?

CT = expand cells in vitro and introduce in patient

TE = organize expanded cells in ECM/Scaffold - implanted into patient
& vascularized.
 Tissue Engineering

• Using well designed scaffolds and optimized

cell growth, the following organs can be
created:
– Skin
– Bone
– Cartilage
– Intestine
Tissue-engineered products contain
mixtures of the following:
• Biological components--cells
• Chemicals
• A non-biological component
• Fibers, plastic, other natural components
 Tissue engineering requires:

Cells

Scaffold Signals
The Ethics and Legislation of
Stem Cell Research

Deborah Burks
Centro de Investigación Principe Felipe
 Stem Cell History
1998 - Researchers first extract stem cells from human embryos.
Thompson et al, H9 and H1.
1999 - First Successful human transplant of insulin-making cells from
cadavers. Edmonton trial.
2001 - President Bush restricts federal funding for embryonic stem-cell
research
2002 - Juvenile Diabetes Research Foundation International creates $20
million fund-raising effort to support stem-cell research
2002 - California ok stem cell research
2004 - Harvard researchers grow stem cells from embryos using private
funding
2004 - Ballot measure for $3 Billion bond for stem cells
2005 - Use of human embryos approved in Spain
Ethics versus Morals

Ethics is a set of moral principles

and a code for behavior that
govern an individual’s actions
with other individuals and within
society.

Morality is what people believe to

be right and good, while ethics is
a critical reflection about morality.
Ethics versus Law

Laws are societal rules or

regulations that are obligatory to
observe.
Laws protect the welfare and safety
of society, resolve conflicts, and are
constantly evolving.

Laws have governed the practice of

medicine for over one hundred of
years.
 Stages of Embryogenesis

Day 2
2-cell embryo Day 3-4
Day 1
Multi-cell embryo
Fertilized egg

Day 5-6
Day 11-14 Blastocyst
Tissue Differentiation
Source of Embryos for Derivation of hESC

• In Vitro Fertilization: Embryo Cryopreserved >5

years

• Embryos Generated and Donated Spefically for

Derivation of hESC

• Embyos that are not suitable for implantation

(Arrested or Defective)
MARCO LEGAL ESPANA

La derivación líneas hESC se

permite en España desde
noviembre del 2003, siempre y
cuando, los embriones donados
previo consentimiento escrito para
dicha derivación, hayan estado
congelados durante al menos 5
años. (Ley 45/2003).
Spanish National Stem Cell Banks
Legislation hESC