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Title: Stem Cell Technology


1
Stem Cell Technology
Presented by
Dr.B.Victor, St.Xavier's College, Palayamkottai
627002 India
2
About the presenter
  • Dr.B.Victor is a highly experienced postgraduate
    biology teacher, recently retired from the
    reputed educational institution St. Xavier s
    College, Palayamkottai, India-627001.
  • He was the dean of sciences and assistant
    controller of examinations.
  • He has more than 32 years of teaching and
    research experience
  • He taught a diversity of courses ranging from
    pre- university to post graduate classes.
  • Send your comments to bonfiliusvictor_at_gmail.com

3
Presentation outline
  • Stem cell characteristics
  • Embryonic stem cells (ESC)
  • Adult Stem cells (ASC)
  • Stem Cell Lines
  • Classification of stem cells
  • Culture and Stem cell therapy
  • Recent Developments

4
Diversity of Human Cells
  • Adult humans consist of more than 200 kinds of
    cells.
  • They are nerve cells (neurons), muscle cells
    (myocytes), skin (epithelial) cells, blood cells
    (erythrocytes, monocytes, lymphocytes, etc.),
    bone cells (osteocytes), and cartilage cells
    (chondrocytes).
  • cells essential for embryonic development but
    not incorporated into the body of the embryo,
    include the extra-embryonic tissues, placenta,
    and umbilical cord.
  • All of these cells are generated from a single,
    totipotent cell, the zygote, or fertilized egg.

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What is a stem cell?
  • A stem cell is a "blank" cell/ precursor cell
    that can give rise to multiple tissue types such
    as a skin, muscle, or nerve cell.
  • A stem cell is essentially the building block of
    the human body.

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Features of Stem Cells
  1. Stem Cells are very unique cells.
  2. Stem Cells have the amazing ability to develop
    into several distinct cell types in the body.
  3. Stem Cells can be used as a repair system for the
    body.
  4. Stem Cells can theoretically divide without
    limit in a living organism in order to replenish
    various types of cells.
  5. When a stem cell divides, each new cell has the
    potential to either remain a stem cell or become
    another type of cell with a more specialized
    function (i.e. a muscle cell, a red blood cell, a
    brain cell, etc.).

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Three unique properties of stem cells
  • Stem cells are capable of dividing and renewing
    themselves for long periods
  • They are unspecialized and they can give rise
    to specialized cell types.
  • A stem cell is "uncommitted," until it receives
    a signal to develop into a specialized cell.

10
Asymmetric division of stem cells
  • Stem cells have the ability to divide
    asymmetrically .
  • One portion of the cell division becomes a
    differentiated cell while the other becomes
    another stem cell.

11
1. Stem cells are unspecialized
  • A stem cell does not have any tissue-specific
    structures that allow it to perform specialized
    functions.
  • A stem cell cannot work with its neighbors to
    pump blood through the body (like a heart muscle
    cell)
  • It cannot carry molecules of oxygen through the
    bloodstream (like a red blood cell) and
  • It cannot fire electrochemical signals to other
    cells that allow the body to move (like a nerve
    cell).

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2.Stem cells are capable of dividing and
renewing themselves for long periods.
  • Stem cells may replicate many times.
  • When cells replicate themselves many times it is
    called proliferation.
  • The stem cells that proliferate for many months
    in the laboratory can yield millions of cells.
  • Stem cells are capable of long-term self-renewal.

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3.Stem cells can give rise to specialized cells
  • When unspecialized stem cells give rise to
    specialized cells, the process is called
    differentiation.
  • There are signals inside and outside cells that
    trigger stem cell differentiation.
  • The internal signals are controlled by a cell's
    genes.
  • The external signals include chemicals secreted
    by other cells, physical contact with neighboring
    cells, and certain molecules in the
    microenvironment

14

4.Stem cells exist in both embryos and adults.
  • In embryos, stem cells function to generate new
    organs and tissues.
  • In adults, they function to replace cells during
    the natural course of cell turnover.

15
Distinguishing Features of Progenitor/Precursor
Cells and Stem Cells.
  • A stem cell is an unspecialized cell that
    develops into a variety of specialized cell
    types.
  • a stem cell divides and gives rise to one
    additional stem cell and a specialized cell.
  • Example a hematopoietic stem cell produce a
    second generation stem cell and a neuron.
  • A progenitor cell (a precursor cell) is
    unspecialized that is capable of undergoing cell
    division and yielding two specialized cells.
  • Example a myeloid progenitor/precursor cell
    undergoing cell division to yield two specialized
    cells (a neutrophil and a red blood cell).

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Stem cell Classes
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Embryonic Type stem cells
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Adult type Stem cells
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Sources of embryonic type stem cells
  •    Embryos - Embryonic stem cells are obtained
    by harvesting living embryos which are generally
    5-7 days old. The removal of embryonic stem cells
    invariably results in the destruction of the
    embryo.
  • Fetuses - Another kind of stem cell, called an
    embryonic germ cell, can be obtained from either
    miscarriages or aborted fetuses.

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Sources of adult type stem cells
  • Umbilical Cords, Placentas and Amniotic Fluid -
    Adult type stem cells can be derived from various
    pregnancy-related tissues.
  • Adult Tissues - In adults, stem cells are present
    within the bone marrow, liver, epidermis, retina,
    skeletal muscle, intestine, brain, dental pulp
    and elsewhere.
  • Cadavers - Neural stem cells have been removed
    from specific areas in post-mortem human brains
    as late as 20 hours following death.

22
Comparison of embryonic and adult stem cells
  • Advantages of  Embryonic Stem Cell   
  • 1. Flexible - appear to have the potential to
    make any cell.  2. Immortal - one embryonic stem
    cell line can potentially provide an endless
    supply of cells with defined characteristics.  3.
    Availability - embryos from in vitro
    fertilization clinics.

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Disadvantages of Embryonic Stem Cell
  1. Difficult to differentiate uniformly and
    homogeneously into a target tissue.
  2. Immunogenic - embryonic stem cells from a random
    embryo donor are likely to be rejected after
    transplantation
  3. Tumorigenic - capable of forming tumors or
    promoting tumor formation.
  4. Destruction of developing human life.

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Advantages of Adult Stem Cell
  1. Adult stem cells from bone marrow and umbilical
    cords appear to be as flexible as the embryonic
    type
  2. Somewhat specialized - inducement may be simpler.
  3. Not immunogenic - recipients who receive the
    products of their own stem cells will not
    experience immune rejection.
  4. Relative ease of procurement - some adult stem
    cells are easy to harvest (skin, muscle, marrow,
    fat)
  5. Non-tumorigenic-tend not to form tumors.
  6. No harm done to the donor.

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Disadvantages of Adult stem cells
  •  1. Limited quantity - can sometimes be
    difficult to obtain in large numbers.  2. Finite
    - may not live as long as embryonic stem cells in
    culture.  3. Less flexible - may be more
    difficult to reprogram to form other tissue types

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Why are adult stem cells preferable to embryonic
stem cells?
  • Adult stem cells are naturally exist in our
    bodies, and they provide a natural repair
    mechanism for many tissues.
  • They belong in the microenvironment of an adult
    body, while embryonic stem cells belong in the
    microenvironment of the early embryo, where they
    tend to cause tumors and immune system reactions.

27
Superior features of ESCs
  • Embryonic stem cells are easier to identify,
    isolate and harvest.
  • There are more of them.
  • They grow more quickly and easily in the lab than
    adult stem cells.
  • They can be more easily manipulated (they are
    more plastic)

28
Classification based on level of differentiation
  • Totipotent
  • Pluripotent
  • Multipotent
  • Unipotent stem cells

29
Types of Stem cells
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Totipotent stem cells
  1. The fertilized egg is said to be totipotent from
    the Latin totus, meaning entire..
  2. It has the potential to generate all the cells
    and tissues that make up an embryo.
  3. It supports embryonic development in utero.

31
Pluripotent stem cells
  • Pluripotent stem cells are descendants of the
    totipotent stem cells of the embryo.
  • These cells develop about four days after
    fertilization
  • They can differentiate into any cell type,
    except for totipotent stem cells and the cells of
    the placenta.

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Pluripotent stem cells
  • Pluri is derived from the Latin plures means
    several or many.
  • Thus, pluripotent cells have the potential to
    give rise to any type of cell.

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Pluripotent stem cells
  • These cells cannot re-create a complete organism
    but differentiate to a large number of mature
    tissue types, for example, brain and muscle.

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Multipotent stem cells
  • Multipotent stem cells are descendents of
    pluripotent stem cells and antecedents of
    specialized cells in particular tissues.
  • For example, hematopoietic stem cells, which are
    found primarily in the bone marrow, give rise to
    all of the cells found in the blood,including red
    blood cells, white blood cells, and platelets.

37
Unipotent stem cell
  • Unipotent stem cell, a term that is usually
    applied to a cell in adult organisms, means that
    the cells in question are capable of
    differentiating along only one lineage.
  • "Uni" is derived from the Latin word unus, which
    means one.

38
Progenitor cells
  • Progenitor cells (or unipotent stem cells) can
    produce only one cell type.
  • For example, erythroid progenitor cells
    differentiate into only red blood cells.

39
Blood is made in the Bone Marrow-Blood Cell
Development
40
Terminally differentiated" cells
  • At the end of the long chain of cell divisions
    are "terminally differentiated" cells, such as a
    liver cell or lung cell, which are permanently
    committed to specific functions.

41
Adult stem cells (ASC)
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Adult stem cells or somatic stem cells
  • Adult stem cells are undifferentiated cells.
  • They are found in small numbers in most adult
    tissues.
  • They can also be extracted from umbilical cord
    blood.
  • They are also called somatic stem cells,
  • They are multipotent in nature.
  • They give rise to a closely related family of
    cells within the tissue.
  • An example is hematopoietic stem cells, which
    form all the various cells in the blood.

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Adult stem cell plasticity and transdifferentiatio
n
  • This ability to differentiate into multiple cell
    types is called plasticity or transdifferentiation
    .

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Differentiation pathways of adult stem cells
  • Neural stem cells in the brain give rise to its
    three major cell types nerve cells (neurons) and
    two categories of non-neuronal cells astrocytes
    and oligodendrocytes.
  • Epithelial stem cells in the lining of the
    digestive tract occur in deep crypts and give
    rise to several cell types absorptive cells,
    goblet cells, Paneth cells, and enteroendocrine
    cells.
  • Skin stem cells occur in the basal layer of the
    epidermis and at the base of hair follicles.
  • The epidermal stem cells give rise to
    keratinocytes, which migrate to the surface of
    the skin and form a protective layer.
  • The follicular stem cells can give rise to both
    the hair follicle and to the epidermis

46
The similarities and differences between
embryonic and adult stem cells
  • Embryonic stem cells can become all cell types
    of the body because they are pluripotent.
  • Adult stem cells are generally limited to
    differentiating into different cell types of
    their tissue of origin.
  • However, some evidence suggests that adult stem
    cell plasticity may exist, increasing the number
    of cell types a given adult stem cell can become.

47
Human embryonic and adult stem cells
  • A potential advantage of using stem cells from an
    adult is that the patient's own cells could be
    expanded in culture and then reintroduced into
    the patient.
  • The use of the patient's own adult stem cells
    would mean that the cells would not be rejected
    by the immune system.
  • Embryonic stem cells from a donor introduced into
    a patient could cause transplant rejection.

48
Umbilical cord stem cells
  • Blood from the placenta and umbilical cord that
    are left over after birth is a rich source of
    hematopoietic stem cells.
  • These so-called umbilical cord stem cells have
    been shown to be able to differentiate into bone
    cells and neurons, as well as the cells lining
    the inside of blood vessels.

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Importance of Cord blood stem cells
  • Cord blood stem cells have been used to treat 70
    different diseases, including leukemia, lymphoma,
    and inherited diseases (of red blood cells, the
    immune system, and certain metabolic
    abnormalities).
  • Cord blood collection is a safe, simple procedure
    that poses no risk to the mother or newborn baby.

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Embryonic Stem Cells (ESC).
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Embryonic Stem Cells
  • Embryonic Stem Cells are derived from embryos
    that develop from eggs that have been fertilized
    in vitro.
  • Embryonic Stem Cells are never derived from eggs
    fertilized inside of a woman's body.
  • The embryos from which Human Embryonic Stem Cells
    are derived are typically four or five days old
    and are a hollow microscopic ball of cells called
    the blastocyst

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Embryonic stem cells (ESC)
  • Embryonic stem cells (ESC), as their name
    suggests, are derived from embryos.
  • Specifically, embryonic stem cells are derived
    from embryos that develop from eggs that have
    been fertilized in vitro donated for research
    purposes with informed consent of the donors.
  • They are not derived from eggs fertilized in a
    woman's body.

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Properties of Embryonic Stem Cells
  • a Derived from the inner cell mass of the
    blastocyst.
  • a Capable of undergoing an unlimited number of
    symmetrical divisions without differentiating
    (long-term self-renewal).
  • Exhibit and maintain a stable, full (diploid),
    normal complement of chromosomes (karyotype).
  • Pluripotent ES cells can give rise to
    differentiated cell types that are derived from
    all three primary germ layers of the embryo
    (endoderm, mesoderm, and ectoderm).

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Potential sources of stem cells are
  • fetal tissue that becomes available after an
    abortion
  • excess embryos from assisted reproductive
    technologies such as commonly used in fertility
    clinics
  • embryos created through in vitro fertilization
    specifically for research purpose, and
  • embryos created asexually as a result of the
    transfer of a human somatic cell nucleus to an
    egg with its own nucleus removed.
  • Other sources of stem cells are those from
    umbilical cord blood, and bone marrow.
  • In addition, neural stem cells, haematopoetic
    stem cells and mesenchymal stem cells can be
    harvested from fetal blood and fetal tissue.

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Cell therapy.
  • Treatment of neural diseases such as Parkinson's
    disease, Huntingtons disease and Alzheimer's
    disease.
  • Stem cells could be used to repair or replace
    damaged neurons.
  • Repair of damaged organs such as the liver and
    pancreas.
  • Treatments for AIDS.

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Stem cell transplantation (SCT)
  • Stem cell transplantation (SCT) is the term now
    used in preference to bone marrow transplantation
    (BMT).
  • When a patient's bone marrow fails to produce new
    blood cells, for whatever reason, he or she will
    develop anaemia, be prone to frequent, persistent
    infections and may develop serious bleeding
    problems.
  • In order to restore blood cell production a
    patient may be given healthy stem cells.

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Therapeutic cloning/ somatic cell nuclear transfer
  • Scientists first remove the nucleus from a normal
    egg cell of a woman. They then extract a nucleus
    from a somatic cell - that is, any body cell
    other than an egg or spermfrom a patient who
    needs an infusion of stem cells to treat a
    disease or injury, and insert the nucleus into
    the egg.
  • The egg, which now contains the patient's genetic
    material, is allowed to divide and soon forms a
    hollow sphere of cells called a blastocyst.
  • Cells from the inner cell mass are isolated and
    used to develop new embryonic stem cell (ESC)
    lines.

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Strategy for therapeutic cloning and tissue
engineering
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Stem cells and cancer treatment
  • Intense chemotherapy damages a persons bone
    marrow, where the stem cells for blood reside.
  • Depleted of a fresh supply of blood cells, the
    patient is left vulnerable to infection, anemia
    and bleeding.
  • These side effects of chemotherapy are often
    treated with a bone marrow transplant.
  • Transplanting bone marrow tissue into a
    chemo-cancer patient may involve hundreds of
    thousands or millions of cells of which only
    two or three may be actual stem cells.
  • It would be much more efficient if you could
    inject a thousand purified stem cells,

63
Therapeutic cloning for tissue repair
  • One human organ, skin, is readily cultured to
    provide replacement tissue for burns victims.
  • Healthy skin cells from the patient can be grown
    rapidly in vitro to provide self-compatible skin
    grafts.

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Is Stem Cell Research Ethical?
  •    Embryonic Stem Cells - always morally
    objectionable, because the human embryo must be
    destroyed in order to harvest its stem cells.  
    Embryonic Germ Cells - morally objectionable when
    utilizing fetal tissue derived from elective
    abortions, but morally acceptable when utilizing
    material from spontaneous abortions
    (miscarriages) if the parents give informed
    consent.   Umbilical Cord Stem Cells - morally
    acceptable, since the umbilical cord is no longer
    required once the delivery has been
    completed.   Placentally-Derived Stem Cells -
    morally acceptable, since the afterbirth is no
    longer required after the delivery has been
    completed.   Adult Stem Cells - morally
    acceptable.

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Sources Consulted
  • Odorico, J.S., Kaufman, D.S., and Thomson, J.A.
    (2001). Multilineage differentiation from human
    embryonic stem cell lines. Stem Cells. 19, 193
    -204.
  • Smith, A.G. (2001). Origins and properties of
    mouse embryonic stem cells. Annu. Rev. Cell. Dev.
    Biol.
  • Thomson, J.A. and Marshall, V.S. (1998). Primate
    embryonic stem cells. Curr. Top. Dev. Biol. 38,
    133-165.
  • Chandross, K.J. and Mezey, E. (2001). Plasticity
    of adult bone marrow stem cells. Mattson, M.P.
    and Van Zant, G. eds. (Greenwich, CT JAI Press).
  • Slack, J.M. (2000). Stem cells in epithelial
    tissues. Science. 287, 1431-1433.

67
Sources Consulted
  • Dzierzak, E., Medvinsky, A., and de Bruijn, M.
    (1998). Qualitative and quantitative aspects of
    haematopoietic cell development in the mammalian
    embryo. Immunol. Today. 19, 228-236.
  • MacKey, M.C. (2001). Cell kinetic status of
    haematopoietic stem cells. Cell. Prolif. 34,
    71-83.
  • J. A. Thomson, et al., 'Embryonic stem cell lines
    derived from human blastocysts', Science, no.
    5391, vol. 282, November 1998, pp. 11457.
  • B. E. Reubinoff, M. F. Pera, C-Y Fong, A.
    Trounson and A. Bongso, 'Embryonic stem cell
    lines from human blastocysts somatic
    differentiation in vitro', Nature Biotechnology,
    vol. 18, pp. 399404, 01 April 2000.

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