Stem cell and its potential application in medicine.

1. Stem Cell and Its
Clinical Implications

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

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

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

5. 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

6. Objectives (Contd)
• Debate for and against stem cell research
• Mention the responsibilities regarding
stem cell issues

7. Stem Cells: Definition
- unspecialized
- self renewal
- can be induced to form specific cell
types

8. 1 stem cell
Self renewal -
maintains the stem cell
pool
4 specialized cells
Differentiation replaces dead or damaged
cells throught the life
Why self-renew AND differentiate?
1 stem cell

9. 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
Don’t show X inactivation
Properties of stem cell….

10. Stem Cell Markers
c-Kit
Oct4 (ATGCAAAT) POU Family Protein
CD34
CD38
Cd44
CD133
Nestin

11. 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.
Laser beam
passes
through one cell
FLUROSCENT
ACTIVATED
CELL SORTING
Stem
cell
SEPARATION OF STEM CELL

12. History of
Stem Cell Research

13. • 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

14. 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

15. 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.

16. SCAN – Stem Cell Action Network
Stem Cell Research Worldwide

17. Global status
• Ongoing debate regarding use of embryos
• United Nations: proposal for a global
policy to ban reproductive cloning only

18. 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

19. 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

20. Types of Stem CellsTypes of Stem Cells
based on potentialbased on potential

21. Types of Stem Cells based on
potential
Stem cellStem cell
typetype DescriptionDescription ExamplesExamples
TotipotentTotipotent
Each cell can developEach cell can develop
into a new individualinto a new individual
Cells from earlyCells from early
(1-3 days)(1-3 days)
embryosembryos
PluripotentPluripotent
Cells can form anyCells can form any
(over 200) cell types(over 200) cell types
Some cells ofSome cells of
blastocyst (5 toblastocyst (5 to
14 days)14 days)
MultipotentMultipotent
Cells differentiated,Cells differentiated,
but can form a numberbut can form a number
of other tissuesof other tissues
Fetal tissue, cordFetal tissue, cord
blood, and adultblood, and adult
stem cellsstem cells

22. This cell
Can form the
Embryo and placenta
This cell
Can just form the
embryo
Fully mature

23. Pluripotent Stem Cells
more potential to become any type of cell

24. Multipotent stem cells
• Multipotent stem
cells – limited in
what the cells can
become

25. Sources of Stem Cell

26. Sources of Stem Cell
• Embryonic stem (ES) cells
• Tissue (Adult) stem cells
• Induced pluripotent stem cells (iPS cells)
• Somatic cell nuclear transfer (therapeutic
cloning)

27. Sources of stem cells
embryonic stem
cells
blastocyst - a very early
embryo
tissue stem cells
fetus, baby and throughout life

28. Embryonic stem (ES) cells:
blastocyst
outer layer of cells
= ‘trophectoderm’
cells inside
= ‘inner cell mass’
embryonic stem cells taken from
the inner cell mass
culture in the lab
to grow more cells
fluid with nutrients

29. Embryonic stem (ES) cells:
embryonic stem cells
PLURIPOTENT
all possible types of specialized
cells
differentiation

30. Tissue stem cells:
muscles
skin
surface of the eye brain
breast
intestines (gut)
bone marrow
testicles

31. Tissue stem cells:
MULTIPOTENT
blood stem cell
found in
bone marrow
differentiation
only specialized types of blood cell:
red blood cells, white blood cells,
platelets

32. Induced pluripotent stem cell
cell from the body
‘genetic reprogramming’
= add certain genes to the cell
induced pluripotent stem (iPS)
cell
behaves like an embryonic stem
cell
Advantage: no need for embryos!
all possible types of
specialized cells
culture iPS cells in the lab
differentiation

33. Induced pluripotent stem cell (Contd)
cell from the body (skin)
genetic reprogramming
pluripotent stem cell
differentiation

34. 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

35. Somatic cell nuclear transfer

36. 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

38. 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!

39. Cloning
There are two VERY different types of cloning:
Reproductive cloning
Used to make two identical
individuals
Very difficult to do
Illegal to do on humans
Molecular cloning
Used to study what a gene
does
Routinely used in the biology
labs
gene 1
gene 2

40. Reproductive cloning
remove nucleus
and take the
rest of the cell
egg
take the nucleus
(containing DNA)
cell from the body
Clone
identical to the individual
that gave the nucleus
Dolly the sheep

41. Molecular cloning
gene 1
gene 2
2) Make a new piece of DNA
gene 1
gene 2
1) Take DNA out of the nucleus
cell 1 cell 2
gene 1 gene 2
3) Put new DNA into a test cell and grow copies
gene 1
cell divides
Daughter cells
contain same DNA:
Genes 1 and 2 have
been cloned
gene 2
insert new DNA

42. Steps of Stem Cell
Therapy

43. 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

44. Steps of Stem Cell Therapy
• Define the problem
Researchers want to replace dead dopamine
neurons with healthy ones

45. • 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

46. • Match The Stem Cell With The Recipient
Needs a good immunonlogical match.
Steps of Stem Cell Therapy (contd.)

47. • 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.)

48. • 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.
Steps of Stem Cell Therapy (contd.)

49. 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

50. • 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

51. Debate for and
against stem cell
research

54. 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.

55. BEFORE
AFTER

56. 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.

58. Stem cell work may bypass
objections

59. Stem-cell work may bypass
objections

60. Stem CellStem Cell
applicationsapplications

61. 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

62. Potential uses cont.
• Biotechnology(drug discovery &
development
–Safety testing of new drugs on
differentiated cell lines
–Screening of potential drugs

63. Potential uses cont.
• Cell based therapies:
–Regenerative therapy to treat
Parkinson’s, heart disease, diabetes
etc
–Stem cells in gene therapy as vehicles
–Stem cells in therapeutic cloning

64. Stem Cell ApplicationsStem Cell Applications
• Tissue repair
- nerve, heart, muscle, organ, skin
• Cancers
• Autoimmune diseases
- diabetes, rheumatoid arthritis, multiple
sclerosis.

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

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

68. 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

70. 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

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

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

73. • window to early brain
development
• Identify critical genes
Down Syndrome

74. Diseases that are treated by stem cells are:
1) Acute Leukemia
• Acute Lymphoblast Leukemia (ALL)
• Acute Myelogenous Leukemia (AML)
• Acute Biphenotypic Leukemia
• Acute Undifferentiated Leukemia
2) Chronic Leukemia
• Chronic Myelogenous Leukemia (CML)
• Chronic Lymphocytic Leukemia (CLL)
• Juvenile Chronic Myelogenous Leukemia (JCML)
• Juvenile Myelomonocytic Leukemia (JMML)
Syndromes
• Myelodysplastic Syndromes
• Amyloidosis
• Chronic Myelomonocytic Leukemia (CMML)
• Refractory Anemia (RA)
• Refractory Anemia with Excess Blasts (RAEB)
• Refractory Anemia with Excess Blasts in Transformation
• (RAEB-T)
• Refractory Anemia with Ringed Sideroblasts (RARS)

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

76. 7) Inherited Immune System Disorders
• Ataxia-Telangiectasia
• Kostmann Syndrome
• Leukocyte Adhesion Deficiency
• DiGeorge Syndrome
• Bare Lymphocyte Syndrome
• Omenn’s 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
• Waldenstrom’s 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

77. Responsibilities
regarding stem cell
issues

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

79. 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.

80. 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

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

82. Mutations can lead to leukemia
Problems with Adult Stem Cells

83. 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/em
bryologie/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/

84. Thank you all