Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
life edu.us
Medical Biotechnology
Lecture 25
Part Ib. Stem Cells - Therapy and
Medical Research
© life_edu
Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
Kimberly Nelson
OnCampus Live
BCH 190, MIC 190, AFS 190, NRS 190, PLS 190
OnLine BCH 190
A Sweeping General Survey on Life and Biotechnology
A Public Access College Course
The University of Rhode Island
Issues in Biotechnology:
Biotechnology, Our Society and Our Future
life
edu.us
Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
life edu.us
BCH 190
Section II.
The Applications of
Biotechnology
A Sweeping General Survey on Life and Biotechnology
© life_edu
The University
of Rhode Island
Stem Cell Research
What is involved?
What is the potential?
What are the downsides?
Who decides?
What, for you, divides our thinking about stem
cell research?
(A) support for the research that may result in
human cloning
(B) uncertain research, not near clinical trials
(C) lack of application, still unproven results
(D) right to life issues
(E) lack of information on the topic
20
15
10
5
0
1
2
3
4
5
Do you support federal funding for embryonic
stem cell research?
(A) yes
(B) no
(C) yes, but with restrictions
30
20
10
0
1
2
3
Therapeutic
vs
Reproductive
Cloning
Embryonic Stem Cells
vs
Adult Stem Cells
vs
iPS Cells
Stem Cell Research
Obama lifts the ban of
Federal funding for
Stem Cell Research
New research will bring
innovation
Stem Cell Basics
Embryonic, Adult
and
Induced Pluripotent
Stem Cells
Stem Cell Basics
What is a Stem Cell?
What Are the Different Types of Stem Cells?
Stem Cell Therapy:
How will this be used?
What is the progress?
What are the controversies?
What is a stem cell?
Characteristics of Stem Cells:

A capacity for extensive self-renewal

Developmental potential
(The ability to differentiate)

The type of stem cell is defined both
by its origin (site, species, age) and by
its differentiation abilities
Examples of types of stem cells based on age
of organism at time of derivation
• Embryonic
• Fetal
• Newborn
• Adult
• iPSc
Stem cells give rise to specialized cells
residing in organs
Stem Cells
Stem Cell Self-renewal
1 stem cell
2 stem cells
Symmetric
Cell
Division
(Proliferation)
Stem Cell Differentiation
1 stem cell
1 stem cell +
1 differentiated
cell
Asymmetric
Cell
Division
(Proliferation)
Stem Cells
Stem Cells
Totipotency
Types of Stem Cells





Embryonic Stem Cells (ES Cells)--Derived from early
embryos (blastocysts) ES cells are totipotent--they can
form all the cell-types in an adult
ES cells are embryonic stem cells and immortal
(unlimited proliferation in dishes)
hES cells are Human embryonic stem cells
Adult Stem Cells - Derived from pluripotent adult
tissues (i.e. bone marrow transplants)
IPS Cells - Induced pluripotent cells are derived from
adult tissues and genetically modified with genes that
override the cell division pathways to become stem cells
capable of then differentiating
Embryonic Stem
Cells
(ES cells)
Preimplantation Development
Zygote
Morula
Extraembryonic
progenitors
Blastocyst
ICM
Preimplantation Development
Zygote
Morula
Extraembryonic
progenitors
Blastocyst
ICM
Derivation of Embryonic Stem Cells
Zygote
ES cells
Morula
Blastocyst
Totipotency of ES cells
Zygote
ES cells
Morula
Blastocyst
How embryonic stem cell lines are produced
Dissociate blastocyst
into single cells
Grow in the presence
of growth factors
Only rarely does a single cell continue to grow indefinitely
How embryonic stem cell lines are validated
Ectoderm
Skin
Neurons
Mesoderm
Muscle
Blood
Bone
Grow cells under different
conditions to stimulate
differentiation into multiple
different cell types
Endoderm
Pancreas
Lung
Liver
Immuno-rejection
Solutions to immuno-rejection
Personalized Stem Cells
Somatic cell nuclear transfer
Unfertilized egg cell
Cell from patient
These cells can also
be genetically
engineered
Remove nucleus
from the egg cell
and replace it with
that of
Use newly formed stem cells
the patient
for therapeutic purposes
Specific to the patient
In addition, ES cells can
be genetically engineered
Stem Cells can be Genetically Modified
Genetic constructs could be used in stem
cell therapy
Promoter
Coding Sequence
Terminator
Your favorite gene
Controlled expression
Cell and developmentally
specific
Insulin
Anti-cancer
Tay-sachs
many others
Stop transcription
Message stability
Nuclear Transfer Stem Cell Therapy
Advantages:
No Rejection (Complete tissuetype match)
Complete Cures
Versatile-Genetic Modification
Disadvantages:
Controversial
Expensive
Requires supply of oocytes
ES Cell Technology is Well Established in
Mice
Zygote
ES cells from mice can be
made genetically engineered
and established in recipients
Use of ES technology is well
established in the mouse
model
Morula
Extraembryonic
progenitors
Blastocyst
ICM
The same Technology Could be Applied to
Humans
Zygote
The same technology could be used to clone a human
Therapeutic
vs
Reproductive
Cloning
Morula
Blastocyst
Embryonic Stem
Extraembryonic
Cells
progenitors
vs
Adult Stem Cells
vs
ICM
iPS Cells
Adult Stem Cells
Types of Stem Cells:

Adult Stem Cells--A small subpopulation of specialized
stem cells present within some adult organs capable of
self renewal
 Adult stem cells are pleuripotent--They have more
limited potential
 Adult stem cells cannot divide indefinitely
Adult Stem Cells
Adult Stem Cells
pluripotency
Some examples of types of stem cells
based on their differentiation abilities
• Hematopoietic stem cells are capable of
differentiation into all mature peripheral blood cells
• Liver stem cells are capable of differentiation into
hepatocytes and cholangiocytes
• Neural stem cells can differentiate into neurons,
astroglia and oligodendrocytes
Adult Stem Cell Therapy
Steps:
1.
2.
3.
4.
5.
Biopsy
In vitro culture of Adult Stem Cells
In vitro expansion or differentiation (If needed)
Genetic modification (as needed)
Engraftment
Bone Marrow Transplantation

Plasticity: The ability of cells to change developmental
fate
Engraftment
Genetic Modification
GFP
(Lymphoid + Myeloid)
Donor Bone Marrow
(Contains Hematopoetic
Stem Cells)
Repopulation of
“Lethally” Irradiated
Host
Adult stem cell differentiation has been demonstrated
Adult Stem Cell Therapy
Advantages:
No Immuno-Rejection (if from self)
Not Controversial
Disadvantages:
Many organs do not contain adult
stem cells
Stem cells may be absent or defective
in the affected organs
Use of Plasticity and
transdifferentiation is unproven
Adult Stem Cells
Self Renewal
(symmetric cell division)
Adult Stem
Cells
Adult Stem Cells
Self Renewal
Adult Stem
Cells
Activation,
Proliferation
(asymmetric cell division)
Adult Organs
Organs With
Adult Stem Cells

Bone Marrow/ Blood
 Skin
 Liver
 Pancreas
 Intestine
 Testes
 Brain ( but limited)
More to come?
Probably!
And these can be
genetically modified
to suit a specific
disease or disorder
Question about Plasticity

Plasticity: The ability of cells to change developmental
fate
Engraftment
GFP
+
(Lymphoid + Myeloid)
Donor Bone Marrow
(Contains Hematopoetic
Stem Cells)
Repopulation of
“Lethally” Irradiated
Host
Neurons
(a few!)
True Plasticity
GFP+
Engrafted
Cells
Donor Bone Marrow
(Contains Hematopoetic
Stem Cells)
Neurons
(a few!)
(Lymphoid + Myeloid)
Induced Pluripotent
Stem Cells (iPS)
Induced Pluripotent
Stem Cells (iPS)
Induced pluripotent stem cells, commonly abbreviated as iPS cells or iPSc, are a
type of pluripotent stem cell artificially derived from a non-pluripotent cell, typically
an adult somatic cell, by inducing a “forced” expression of certain genes that regulate
the cell division cycle thus making them immortal.
Induced Pluripotent Stem Cells are believed to be identical to natural pluripotent stem
cells, such as embryonic stem cells in many respects, such as the expression of certain
stem cell genes and proteins, chromatin methylation patterns, doubling time, embryo
formation, teratoma formation, viable chimera formation, and potency and
differentiability, but the full extent of their relation to natural pluripotent stem cells is
still being assessed.
IPScs were first produced in 2006 from mouse cells and in 2007 from human cells.
This has been cited as an important advancement in stem cell research, as it may allow
researchers to obtain pluripotent stem cells, which are important in research and
potentially have therapeutic uses, without the controversial use of embryos.
Induced Pluripotent
Stem Cell Basics
A scheme of the generation of induced pluripotent stem (iPS) cells. (1) Isolate and
culture donor cells. (2) Transfect stem cell-associated genes into the cells by viral
vectors. Red cells indicate the cells expressing the exogenous genes. (3) Harvest and
culture the cells according to ES cell culture, using mitotically inactivated feeder cells
(4) A small subset of the transfected cells become iPS cells and generate ES-like
colonies.
INDUCED PLURIPOTENT Stem Cell Lines Derived from Human Somatic Cells
Junying Yu 1*, Maxim A. Vodyanik 2, Kim Smuga-Otto 1, Jessica Antosiewicz-Bourget 1, Jennifer L Frane 3, Shulan Tian 4, Jeff Nie 4, Gudrun A.
Jonsdottir 4, Victor Ruotti 4, Ron Stewart 4, Igor I. Slukvin 5, James A. Thomson 6
Science DOI: 10.1126/science.1151526
Stem Cell Therapy:
Recent Progress
Stem Cell Therapy
Human Degenerative Disorders:
Many diseases are characterized by organ failure or cell loss:

Degenerative disease states (unknown etiology or autoimmune
disease)

Age-related degenerative disease

Cell or organ destruction due to genetic or infectious disease
Stem Cell Therapy:

Stem Cell Therapy is the replenishment of cells or
organs lost to degenerative disease or loss of function
with new cells derived from stem cells

Stem Cell Therapy is Experimental
(Except Bone Marrow Transplantation)

Stem Cell therapy should be free of pharmaceutical
toxicity

Cellular Pharmaceutical
Stem Cell Therapy
Stem Cell Therapy offers a method to treat
degenerative disorders
A Cellular Pharmaceutical
Diseases amenable to Stem Cell Therapy






Diabetes
Spinal chord injury
Alzheimer’s Disease
Parkinson’s Disease
Heart Disease
Many more…..but not overnight
Stem Cell Therapy
Established Stem Cell Therapies:
Bone Marrow Transplantation--Restores blood system
with hematopoetic stem cells (1950s)
(many variations)
Simplest Diseases to cure next involve secretory cells:
 Parkinson’s Disease--Dopamine Secretion
 Type I Diabetes--Insulin Secretion
(Type II is difficult due to insulin insensitivity)
in vitro:
ES cell Therapy
(Parkinson’s Disease)
in vivo:
Dopamine
Production
Restored
ES cells
Engraftment
Dopaminergic
Neurons
Parkinson’s Disease-Experimental
Therapy
From Lindvall et al., Nat. Med. 2004, S42-S50.
Stem Cell Therapy
Criticisms and Controversies
Embryogenic Stem Cell
Criticisms
I. Destruction of human embryos
II. A slippery slope to:
• Human Genetic Engineering
• Human cloning
Somatic cell nuclear transfer techniques are
the same as those that could be applied to
cloning
Unfertilized egg cell
Cell from patient
These cells can also
be genetically
engineered
Remove nucleus
from the egg cell
and replace it with
that of
Use newly formed stem cells
the patient
for therapeutic purposes
Specific to the patient
In addition, ES cells can
be genetically engineered
Where is DNA in a cell?
• All of the information for each protein in a cell is in it’s DNA
• All of the information that codes for a complete organism is in DNA
7. Induced Pluripotent Stem Cells (iPCs) are made by the
introduction of genes that cause the normal cell cycle to
remain in the cell division mode, ensuring that they will divide
in culture indefinitely. Two or more cloned genes are inserted
into the genome of adult differentiated cells; these genes are
similar to those involved with tumor genesis that override
normal cell division. These cells can then be triggered to
become any adult cell type, thereby obviating the controversy
about using cells derived from human embryos. The process
of inserting genes into cells is called:
(A)
(B)
(C)
(D)
(E)
eugenics
carcinogenics
transgenics
genomics
idiomics
8. Stem cell research in humans is controversial, in part,
because of the so-called ‘slippery slope’ argument. It is
suggested that the same technology that is developed for hES
stem cells and therapeutic cloning could easily be extended to:
(A)
(B)
(C)
(D)
(E)
determining who can have children
reproductive cloning of humans
increased abortions
a policy of ethnic cleansing
the creation of bioweapons
9. The defining characteristic(s) of adult stem cells are that:
(A) they are pluripotent - they have a limited potential to
differentiate to other cell types
(B) all of these answers are appropriate
(C) they are seen as a potential source for therapeutic
purposes that will not be controversial
(D) they are unable to divide indefinitely
(E) they can be genetically engineered to carry and express
foreign genes
10. Adult Stem Cells are:
(A) outlawed in every country in the world as part of a wide
ban on cloning
(B) from a small subpopulation of specialized stem cells
present within some adult organs and capable of self renewal
and limited capacity to differentiate
(C) an artifact that was famous for being fraudulently
reported
(D) unable to differentiate into any other cell type
(E) unable to be genetically altered the way hES cells are
11. Degenerative diseases that result in cell death or function,
such as Parkinson’s and Alzheimer’s, injuries that may result
in damages cells and tissues, such as spinal cord injuries, and
the lack of available human donor organs for transplant has
motivated scientists to investigate new ways to replace the
functions of diseased organs. Promising approaches to these
problems that have received considerable research are:
(A)
(B)
(C)
(D)
(E)
xenotransplantation and tissue engineering
all of these approaches
gene therapy
creation of artificial biomedical devices
stem cells
12. Human Embryonic Stem Cells (hES Cells) are:
(A) derived from adult bone marrow as leukemia treatments
(B) derived from early (five day old) embryos (blastocysts)
(C) outlawed in every country in the world
(D) made with great ease at the Seoul National University in
South Korea
(E) were funded through vast amount of Federal dollars in the
US from 2001-2008
13. The history behind the development and therapeutic
applications for adult stem cells includes that they:
(A) have been used to reproductively clone primates and in
one case an adult human being
(B) have been derived from every tissue type in the human
body and have been shown to have equal developmental
plasticity compared with hES cells
(C) have been shown to be more useful than embryonic stem
cells for any applications, demonstrating the importance of
hES cells
(D) have been used to treat leukemia using bone marrow since
1956 and have been used successfully without controversy for
many years
(E) in spite of their great promise, because of serious errors
will never be successful in clinical trials to treat any disease
and have resulted in many fatalities
14. Many diseases are characterized by organ failure or cell
loss. These types of disease are attractive candidates for stem
cell therapy and include:
(A) disease states that require gene therapy
(B) only degenerative disease of unknown etiology or
autoimmune disease
(C) only age-related degenerative diseases, such as
Parkinson’s and Alzheimer’s
(D) only cell or organ destruction due to genetic,
environmental or infectious disease, such as diabetes
(E) all of the disease states described in the other answers
15. Stem cells give rise to specialized cells residing in organs.
One of the hallmark characteristics of stem cells is a prolonged
capacity for self-renewal. Another hallmark characteristic of
stem cells is their:
(A) origin exclusively in embryos that are five days old
(B) developmental potential (their ability to differentiate in
any cell type)
(C) inability to be genetically engineered
(D) inability to grow in culture
(E) ability to be cultured back to cloned human beings
16. The production of human embryonic stem cells is
controversial because:
(A) they have been used to clone President George W. Bush
(B) a three month old fetus must be aborted in order to
produce them
(C) a blastocyst stage embryo usually must be ‘dissociated’ to
recovery ICM (Inner Cell Mass) cells in order to produce them
in culture
(D) these types of cells can be used only for reproductive
cloning
(E) they have been shown to cause delusions in NIH
researchers about the prospects of receiving federal funding
for this important work
17. Stem Cell Research:
(A) was banned from receiving federal funding in the US in
August 2001 under the Bush Administration until recently but
is still in contention by many public opponents
(B) has resulted in human cloning by a group in the
Netherlands as reported on the internet
(C) is an established science since first applied to a dog skull
bone in the 1600s
(D) has no scientific basis and because of this should not
receive federal funding
(E) is a scientific ruse developed by a fraudulent researcher in
South Korea named Dr. Woo Suk Wong