Stem Cell and Its

Clinical Implications

Presented by:
DR.Mahfujun Nahar
MS Phase-A Resident
Department of Anatomy
BSMMU

Guided by
Dr. Nahid Farhana Amin
Assistant Professor
Department of Anatomy
BSMMU

Objectives
Define stem cell
Outline brief history of stem cell research
Mention the types of stem cells based on
potential

Objectives (contd)
Outline the sources of stem cell
Explain the steps of stem cell therapy
Discuss the health problems that might be
treated by stem cells

Objectives (Contd)

Debate for and against stem cell research

Mention the responsibilities regarding
stem cell issues

Stem Cells: Definition

- unspecialized
- self renewal
- can be induced to form specific cell
types

Why self-renew AND differentiate?

1 stem cell

1 stem cell

4 specialized cells

Self renewal maintains the stem cell

pool

Differentiation replaces dead or damaged

cells throught the life

Properties of stem cell.

Clonogenic, a single ES cell gives rise to a colony of
genetically identical cells, which have the same properties
as the original cell
Expresses the transcription factor Oct-4
Can be induced to continue proliferating or
to differentiate
Lacks the G1 checkpoint in the cell cycle
ES cells spend most of their life cycle in S phase
Dont show X inactivation

Stem Cell Markers

c-Kit
Oct4 (ATGCAAAT) POU Family Protein
CD34
CD38
Cd44
CD133
Nestin

SEPARATION OF STEM CELL

Stem
cell

FLUROSCENT
ACTIVATED
CELL SORTING
Laser beam
passes
through one cell

Cells in suspension are tagged

with fluorescent markers
specific for undifferentiated
stem cell
Labeled cells are sent under
pressure through a small nozzle
and pass through an electric
field
A cell generates a negative
charge
if it fluoresces and a positive
charge
if it does not.

History of
Stem Cell Research

History of
Stem Cell Research
In 1998, James Thomson
isolated stem cells from the
inner cell mass of the early
embryo.
In 1998, John Gearhart derived
human embryonic germ cells
from fetal gonadal tissue
(primordial germ cells).

History of Stem Cell Research (Contd)

1999 - First Successful human transplant of
insulin-making cells from cadavers
2001 - President Bush restricted federal
funding for embryonic stem-cell
research

History of Stem Cell Research (Contd)

2004 - Harvard researchers grow stem cells
from embryos using private funding.
Asia, Japan, South Korea and Singapore is
moving forwards on stem cell research.

Stem Cell Research Worldwide

SCAN Stem Cell Action Network

Global status
Ongoing debate regarding use of embryos

United Nations: proposal for a global

policy to ban reproductive cloning only

Debate in US
Federal funding available for research using
the Bush lines only
ES cell lines from 8/9/01
Disadvantage of Bush stem cell lines:
may have mutations or infections
Private companies continue to pursue stem
cell research
therapeutic cloniing mainly

Stem cell research in other

countries
Great Britain
Therapeutic cloning , use of excess
embryos & creation of embryos allowed
France
Reproductive and therapeutic cloning
banned
Germany
Use of excess embryos and creation of
embryos banned

Types of Stem Cells

based on potential

Types of Stem Cells based on

potential

Stem cell
type

Description

Examples
Cells from early
(1-3 days)
embryos

Totipotent

Each cell can develop

into a new individual

Pluripotent

Cells can form any

(over 200) cell types

Multipotent

Cells differentiated,
Fetal tissue, cord
but can form a number blood, and adult
of other tissues
stem cells

Some cells of
blastocyst (5 to
14 days)

This cell
Can form the
Embryo and placenta

This cell
Can just form the
embryo

Totipotent
Pluripotent
Multipotent
Fully mature

Pluripotent Stem Cells

more potential to become any type of cell

Multipotent stem cells

Multipotent stem
cells limited in
what the cells can
become

Sources of Stem Cell

Sources of Stem Cell

Embryonic stem (ES) cells
Tissue (Adult) stem cells
Induced pluripotent stem cells (iPS cells)
Somatic cell nuclear transfer (therapeutic
cloning)

Sources of stem cells

embryonic stem
cells
blastocyst - a very early
embryo

tissue stem cells

fetus, baby and throughout life

Embryonic stem (ES) cells:

blastocyst
cells inside
= inner cell mass

embryonic stem cells taken from

the inner cell mass
outer layer of cells
= trophectoderm

fluid with nutrients

culture in the lab

to grow more cells

Embryonic stem (ES) cells:

differentiation
embryonic stem cells

PLURIPOTENT

all possible types of specialized

cells

Tissue stem cells:

surface of the eye

skin

testicles

brain
breast

intestines (gut)

bone marrow
muscles

Tissue stem cells:

blood stem cell

differentiation
found in
bone marrow

MULTIPOTENT

only specialized types of blood cell:

red blood cells, white blood cells,
platelets

Induced pluripotent stem cell

genetic reprogramming
= add certain genes to the cell
cell from the body

induced pluripotent stem (iPS)

cell
behaves like an embryonic stem
cell

differentiation
culture iPS cells in the lab

Advantage: no need for embryos!

all possible types of

specialized cells

Induced pluripotent stem cell (Contd)

genetic reprogramming
pluripotent stem cell

cell from the body (skin)

differentiation

Somatic cell nuclear transfer

A nucleus from an adult donor cell is inserted
into a recipient egg cell from which the nucleus
has been removed
The resulting cell is then stimulated to
divide as a zygote later forming embryo
genetically identical to the adult donor
cell

Somatic cell nuclear transfer

Goals of therapeutic cloning

Use embryo as source for ES cells
Use ES cells to generate an organ with
genetic markers identical to the patient
Correct genetic error in ESC in blastula stage

Pitfalls of therapeutic cloning

Large number of eggs needed for SCNT
To harvest large number of eggs:
excessive hormone treatment may induce
high rate of ovulation
will carry species-specific mitochondrial
genes
Mixing species is reason for concern!

Cloning
There are two VERY different types of cloning:
Reproductive cloning

Molecular cloning
gene 1
gene
2

Used to make two identical

individuals

Used to study what a gene

does

Very difficult to do

Routinely used in the biology

labs

Illegal to do on humans

Reproductive cloning

cell from the body

take the nucleus

(containing DNA)

Clone
identical to the individual
that gave the nucleus
Dolly the sheep

egg
remove nucleus
and take the
rest of the cell

Molecular cloning
Take DNA out of the nucleus
gene
1

cell 1

gene 2
cell 2

) Make a new piece of DNA

gene 1

gene 1

gene
2
3) Put new DNA into a test cell and grow
copies
gene
1

gene
2
insert new
DNA

cell divides

gene 2

Daughter cells
contain same DNA:
Genes 1 and 2 have
been cloned

Steps of Stem Cell

Therapy

Steps of Stem Cell Therapy

Defining the problem
Finding The Right Type of Stem Cell
Match The Stem Cell With The Recipient
Put the stem cells in the right place
Make The Transplanted Stem Cells Perform

Steps of Stem Cell Therapy

Define the problem
Researchers want to replace dead dopamine
neurons with healthy ones

Steps of Stem Cell Therapy

Finding The Right Type of
Stem Cell
Blastocyst stem cells?
At the time, unable to
differentiate into neurons
Fetal stem cells?, Excellent
candidates, ethical problems
Adult stem cells?, Hard to get,
too little known

Steps of Stem Cell Therapy (contd.)

Match The Stem Cell With The Recipient
Needs a good immunonlogical match.

Steps of Stem Cell Therapy (contd.)

Put the stem cells in the right place

Surgical procedure usually required.

Small holes drilled in the skull, cells
injected with a needle.

Steps of Stem Cell Therapy (contd.)

Make The Transplanted Stem Cells Perform
There was no guarantee how the transplanted cells
would behave. If they did not respond to the proper
signals from their environment, they might have
malfunctioned or died.

Cell Culture Techniques for ESC

Isolate & transfer of inner cell mass into
plastic culture dish that contains culture
medium
Cells divide and spread
Inner surface of culture dish is typically
coated with mouse embryonic skin cells
that have been treated so they will not
divide

 This coating is called feeder layer:

provide ES cells with a sticky surface for
attachment and release nutrients
There are methods for growing
embryonic stem cells without mouse
feeder cells
ES cells are removed gently and plated
into several different culture plates

Debate for and

against stem cell
research

Pro-choice people
Utilitarianism- destruction of smaller
group for the sake of a larger group is
justifiable.
lead to significant information about the
cause, new treatment possibilities, and
potential cure for many diseases.

AFTER

BEFORE

Opinions against stem cell research

Stem cells are taken from a human
blastocyst, which is then destroyed. This
amounts to murder.
There is a risk of commercial exploitation
of the human participants in ESCR.

Stem cell work may bypass

objections

Stem-cell work may bypass

objections

Stem Cell
applications

Potential Uses of Stem Cells

Basic research clarification of complex
events such as
Molecular mechanisms for gene control
Role of signals in gene expression &
differentiation of the stem cell
Stem cell theory of cancer

Potential uses cont.

Biotechnology(drug discovery &
development
Safety testing of new drugs on
differentiated cell lines
Screening of potential drugs

Potential uses cont.

Cell based therapies:
Regenerative therapy to treat
Parkinsons, heart disease, diabetes
etc
Stem cells in gene therapy as vehicles
Stem cells in therapeutic cloning
Stem cells in cancer

Stem Cell Applications

Tissue repair
- nerve, heart, muscle, organ, skin
Cancers
Autoimmune diseases
- diabetes, rheumatoid arthritis, multiple
sclerosis.

Tissue Repair
Regenerate spinal cord, heart
tissue or any other major tissue in
the body.

Replace Skin

http://www.youtube.com/watch?feature=player_embedded&v=eXO_ApjKPaI

Heart Disease
Adult bone marrow stem cells injected
into the hearts are believed to improve
cardiac function in victims of heart failure
or heart attack

Leukemia and Cancer

Leukemia patients treated with stem
cells emerge free of disease.
Stem cells have also reduces
pancreatic cancers in some patients.

Proliferation of white cells

Rheumatoid Arthritis
Adult Stem Cells may be helpful in
jumpstarting repair of eroded cartilage.

Type I Diabetes
Embryonic Stems Cells might be trained
to become pancreatic islets cells needed
to secrete insulin.

Down Syndrome
window to early brain
development
Identify critical genes

eases that are treated by stem cells are:

cute Leukemia
ute Lymphoblast Leukemia (ALL)
ute Myelogenous Leukemia (AML)
ute Biphenotypic Leukemia
ute Undifferentiated Leukemia
hronic Leukemia
ronic Myelogenous Leukemia (CML)
ronic Lymphocytic Leukemia (CLL)
venile Chronic Myelogenous Leukemia (JCML)
venile Myelomonocytic Leukemia (JMML)
dromes
yelodysplastic Syndromes
myloidosis
ronic Myelomonocytic Leukemia (CMML)

Disorders
5) Inherited Metabolic Disorders
1) Stem Cell Disorders
Mucopolysaccharidoses (MPS)
Aplastic Anemia (Severe)
Hurlers Syndrome (MPS-IH)
Fanconi Anemia
Scheie Syndrome (MPS-IS)
Paroxysmal Nocturnal Hemoglobinuria Hunters Syndrome (MPS-II)
Congenital Cytopenia
Sanfilippo Syndrome (MPS-III)
Dyskeratosis Congenita
Morquio Syndrome (MPS-IV)
2) Myeloproliferative Disorders
Maroteaux-Lamy Syndrome (MPS-VI)
Acute Myelofibrosis
Sly Syndrome, Beta-Glucuronidase Deficiency
Agnogenic Myeloid Metaplasia
Adrenoleukodystrophy
Polycythemia Vera
Mucolipidosis II (I-cell Disease)
Essential Thrombocythemia
Krabbe Disease
3) Lymphoproliferative Disorders
Gauchers Disease
Non-Hodgkins Lymphoma
Niemann-Pick Disease
Hodgkins disease
Wolman Disease
Prolymphocytic Leukemia
Metachromatic Leukodystrophy
4) Phagocyte Disorders
6) Histiocytic Disorders
Chediak-Higashi Syndrome
Familial Erythrophagocytic Lymphohistiocytosis
Chronic Granulomatous Disease
Histiocytosis-X
Neutrophil Actin Deficiency
Hemophagocytosis
Reticular Dysgenesis
Langerhans Cell Histiocytosis

7) Inherited Immune System Disorders

Ataxia-Telangiectasia
Kostmann Syndrome
Leukocyte Adhesion Deficiency
DiGeorge Syndrome
Bare Lymphocyte Syndrome
Omenns Syndrome
Severe Combined Immunodeficiency
SCID with Adenosine Deaminase
Deficiency
Absence of T & B Cells SCID
Absence of T Cells, Normal B Cell SCID
Common Variable Immunodeficiency
Wiskott-Aldrich Syndrome
X-Linked Lymphoproliferative Disorder
Other Inherited Disorders
Lesch-Nyhan Syndrome
Cartilage-Hair Hypoplasia
Glanzmann Thrombasthenia
Osteopetrosis
Adrenoleukodystrophy
Ceroid Lipofuscinosis
Congenital Erythropoietic Porphyria
Sandhoff Disease

9) Plasma Cell Disorders

Multiple Myeloma
Plasma Cell Leukemia
Waldenstroms Macroglobulinemia
Amyloidosis
Abnormalities
1) Inherited Platelet Abnormalities
Congenital Thrombocytopenia
2) Inherited Erythrocyte Abnormalities
Beta Thalassemia Major
Sickle Cell Disease
Blackfan-Diamond Anemia
Pure Red Cell Aplasia
Other Malignancies
Ewing Sarcoma
Neuroblastoma
Renal Cell Carcinoma
Retinoblastoma
Brain tumor
Ovarian Cancer
Small Cell Lung Cancer
Testicular Cancer

Responsibilities
regarding stem cell
issues

Responsibilities regarding stem cell

issues
Become informed
The facts about stem cell research and its
curative potential.
www.stemcellfunding.org
www.stemcellaction.org

Responsibilities regarding stem

cell issues
(contd.)

Inform others
Contact patient and community groups
and offer to give a presentation like this
one. Organize a house party to help
spread the word.
Collect email addresses of supporters to
be added to mailing list.

Responsibilities regarding stem

cell issues (contd.)
Inform others
Arrange to meet with your political
representatives to discuss their support for
stem cell research
Find other like-minded people and work
together
Invite friends, colleagues, and caretakers
of patients to become involved

Technical Challenges
Source - Cell lines may have mutations
Delivery to target areas
Prevention of rejection
Suppressing tumors

Problems with Adult Stem Cells

Mutations can lead to leukemia

REFERENCES
Stem cells in class; Badran, Shahira; Bunker Hill Community
College, 2007, Boston Museum of Science Biotechnology
Symposium
Stem cells & Cloning Stem cells & Cloning; David A. Prentice,
Benjamin Cummings, 2003
http://stemcells.nih.gov/info/scireport/2001report.htm
http://www.ulb.ac.be/sciences/biodic/biodic/images/bio_animale/emb
ryologie/fecondation/baefec_01_01.jpg
http://www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=mboc4.figgrp.3748
http://www.news.wisc.edu/packages/stemcells/illustration.html
http://www.drugs.com/enc/images/images/en/17010.jpg
http://embryology.med.unsw.edu.au/Notes/placenta.htm
http://www.dnalc.org/stemcells.html
http://gslc.genetics.utah.edu/units/stemcells/

Thank you all