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Dr.T.V.Rao MD
STEM CELLSADVANCES IN SCIENCE
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STEM CELL DEFINITION
A cell that has the
ability to
continuously divideand differentiate
(develop) into
various otherkind(s) of
cells/tissues
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DR.T.V.RAO MD
WHAT MAKES A CELL A STEM CELL?
Plasticity: A stem cell is plastic, meaning it candevelop into another type of cell.
Differentiation: The process where a stem cellspecializes or develops into another type ofcell.
Self-Renewal: a stem cell can divide (renewitself) indefinitely(go through mitosis) andwithout always developing into another cell.
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STEM CELL HISTORY
1998 - Researchers first extract stem cells from human embryos
1999 - First Successful human transplant of insulin-making cellsfrom cadavers
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
2003?? - California ok stem cell research
2004 - Harvard researchers grow stem cells from embryos usingprivate funding
2004 - Ballot measure for $3 Billion bond for stem cells
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HISTORY OF HUMAN EMBRYONICSTEM CELL RESEARCH
In 1998, James Thomson(University ofWisconsin-Madison) isolated cells from the innercell mass of the blastocyst, and developed the first
human embryonic stem cell line in culture.
Isolate inner cell mass
Culture cells
Inner cell mass(forms fetus)
Day 5-6Blastocyst
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IN 1998, JAMES THOMSON (UNIVERSITY OF
WISCONSIN-MADISON) ISOLATED CELLS FROM
THE INNER CELL MASS OF THE EARLY EMBRYO,
AND DEVELOPED THE FIRST HUMAN EMBRYONICSTEM CELL LINES,
HISTORY OF HUMAN EMBRYONIC STEM CELLRESEARCH
In 1998, John Gearhart (JohnsHopkins University) derived human
embryonic germ cells from cells infetal gonadal tissue (primordialgerm cells).
Pluripotent stem cell lines were
developed from both sourcesDR.T.V.RAO MD 6
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WHAT ARE STEM CELLS?
Stem cells are the raw
material from which all of
the bodys mature,
differentiated cells aremade. Stem cells give
rise to brain cells, nerve
cells, heart cells,pancreatic cells, etc.
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DR.T.V.RAO MD
MAJOR TYPES OF STEM CELLS
Embryonic Stem Cells
Totipotent : cell can
develop into all celltypes
Immortal: can self-renew indefinitely
Plentiful
Adult Stem Cells
Multipotent: cell can develop
into a few cell types but notall
Located in few organs or
may be unidentified
Hard to find
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STEM CELL ARE DYNAMIC
Are undifferentiatedmastercell that do notyet have a specificfunction
Can change to one orseveral different cell types(differentiate) under properconditions
Can undergo unlimited celldivision, self-renewal)
Stem cell
Stem cell Specialized cell(e.g., white blood cell)
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ONE CELL - SEVERAL LINEAGES
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EMBRYOGENESIS ANDDIFFERENTIATION
Specific regions of the embryo giverise to the specific organ systems Ectodermgenerates the outer layer of the embryo
and produces the surface layer (epidermis) of the skinand forms the nerves
Endoderm becomes the innermost layer of the
embryo and produces the digestive tube and itsassociated organs (including the lungs)
Mesodermbecomes sandwiched between theectoderm and endoderm and generates the blood,
heart, kidney, gonads, bones, and connective tissues.DR.T.V.RAO MD 11
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STAGES OF DEVELOPMENT
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EARLY HUMAN DEVELOPMENT
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AN OVERVIEW OF EARLY DEVELOPMENT
MODELED WITH PLAY-DOUGH
Fertilized egg
Totipotentstem cells
Totipotent: Can become any cell inbody or placenta
Fate Decision
Pluripotentstem cells(3-5 days old)
Blastocyst
Pluripotent: Can become any cell in
body
Implantation
Fate Decision
Gastrulation (day 14) leads toPrimary Germ CellsEndoderm (inner)digestive tract, resp. trackMesoderm (middle)bones, blood cells, heartEctoderm (outer)skin, CNS
Multipotent: Can become any cell
within a specific germ layer or celllineage
Embryonic stem cells come from inner cell mass of blastocyst.
Implantation
Fate Decision
Gastrulation (day 14) leads toPrimary Germ CellsEndoderm (inner)digestive tract, resp. trackMesoderm (middle)bones, blood cells, heartEctoderm (outer)skin, CNS
Multipotent: Can become any cell
within a specific germ layer or celllineage
Multipotent
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Somatic Cell
Endoderm Ectoderm Mesoderm
LiverIntestinePancreas
SkinHair
Nerves
BloodMuscleBone
Cartilage
Somaticstem cell
Germ stemcell
Primitivegerm cell
Gametes
PluripotentEmbryonal stem cell
Totipotentstem cells
Pluripotentstem cells
Adult Multipotent
stem cell
Courtesy of Dr F. Prosper Cardoso.DR.T.V.RAO MD 15
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BONE MARROW STEM CELLS
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HOW TO DERIVE AN EMBRYONIC STEM CELL LINE?
Inner cell mass
Day 5-6Blastocyst
Isolate inner cell mass(destroys embryo)ETHICS?
Culture cells
A stem cell line iscomposed of a
population of cellsthat can replicatethemselves forlong periods oftime in vitro(out of the body) An embryonic
stem cellcloneDR.T.V.RAO MD 17
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THE SCIENCE OF STEM CELLS
Stem cells have the ability to continually reproducethemselves while maintaining the capacity to giverise to other more specialized cells.
Stem cells are found at all stages of development,from embryonic stem (ES) cells that candifferentiate into all specialized cells found in thehuman body, to adult stem cells capable of
regenerating their tissue of origin.
Stem cells occur from the earliest stages ofdevelopment and provide the starting material
for every organ and tissues.DR.T.V.RAO MD 18
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EMBRYONIC STEM (ES) CELLS
ES cells arefound at the
blastocyst stage,four to five daysafter the union of
the sperm andegg, before theembryo implants
in the uterus.DR.T.V.RAO MD 19
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ES CELLS ARE "PLURIPOTENT" - I.E. CAPABLE OFFORMING EMBRYONIC TISSUES
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Stem cells may be derived
from autologous,
allogeneic or xenogeneic
sources. Histocompatibility
is prerequisite fortransplantation of
allogeneic stem cells. Fatal
tissue is the best current
tissue source for humanneural stem cells, however
ethical issues are a major
concern.
SOURCE OF STEM CELLS
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C SO C O S
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PLACENTA A SOURCE OF STEMCELLS
Placental stem cells,
like umbilical cord
blood and bone
marrow stem cells,can be used to cure
chronic blood-related
disorders such assickle cell disease,
Thalassemia, and
leukaemia.DR.T.V.RAO MD 23
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REPORT OFPRELIMINARY
RESULTS OFTRANSPLANTATIONUSING PARTIALLY HLA-MISMATCHEDPLACENTAL BLOODFROM UNRELATEDDONORS.
JOANNE KURTZBERG,M.D.ET AL
Placental Blood as a Source of Hematopoietic Stem
Cells for Transplantation into Unrelated Recipients
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UMBILICAL CORD BLOOD STEM CELLTRANSPLANT
Umbilical cord blood
stem cell transplants are
less prone to rejection
than either bone marrowor peripheral blood stem
cells. This is probably
because the cells have
not yet developed thefeatures that can be
recognized and attacked
by the recipient's immune
system DR.T.V.RAO MD 25
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KINDS OF STEM CELLSStem cell type Description Examples
Totipotent Each cell can develop into a
new individual
Cells from early (1-3
days) embryos
Pluripotent Cells can form any (over 200)
cell types
Some cells of blastocyst
(5 to 14 days)
Multipotent Cells differentiated, but can
form a number of other tissues
Fetal tissue, cord blood,
and adult stem cells
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They have the potential toreplace cell tissue that hasbeen damaged ordestroyed by severeillnesses.
They can replicatethemselves over and overfor a very long time.
Understanding how stem
cells develop into healthyand diseased cells willassist the search for cures.
WHATS SO SPECIAL ABOUT STEM CELLS?
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Embryonic(also calledpluripotent) stem cellsare capable of developinginto all the cell types of the
body.
Adult stem cells are lessversatile and more difficultto identify, isolate, and
purify.
TWO KINDS OF STEM CELLS
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EMBRYONIC STEM CELLSEmbryos are formedin labs that helpcouples get
pregnant. An eggand sperm fertilize azygote and areinserted into awomans uterus to
develop into anembryo and thenfetus.
http://biodidac.bio.uottawa.ca/thumbnails/filedet.htm?File_name=HUMN150B&File_type=GIF
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STAGES OF EMBRYOGENESIS
Day 1Fertilized egg
Day 22-cell embryo Day 3-4Multi-cell embryo
Day 5-6BlastocystDay 11-14
Tissue DifferentiationDR.T.V.RAO MD 30
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DERIVATION AND USE OF EMBRYONIC STEMCELL LINES
Isolate inner cell mass(destroys embryo)
Heart muscleKidney
Liver
Special sauce(largely unknown)
Day 5-6Blastocyst
Inner cells
(forms fetus)
Outer cells(forms placenta)
Heart
repaired
Culture cells
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EMBRYONIC STEM CELLS
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EMBRYONIC STEM CELLS:Researchers extract stem cells from a 5-7 days oldblastocyst.
Stem cells can divide in culture to form more of their own kind,
thereby creating a stem cell line.
The research aims to induce these cells to generate healthytissue needed by patients.
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Adult stem cells are calledsomatic or body stem cells.Some people call these adultbecause they are found afteran embryo develops into a
fetus and are no longer anembryonic stem cell. Not onlyadults have adult stemcells! Some organs arebelieved to lack stem cells
and these cells grow andreplace dead cells withmitosis.
ADULT STEM CELLS
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Adult stem cells develop
into a few cell types. These
Multipotent cells are usedin bone-marrow transplants
and will develop into all theblood cells. These cells are
important, but some organs
may not have adult stem
cells and these cells can bedifficult to find
ADULT STEM CELLS
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AUTOLOGOUS STEM CELLS Sources of the patient's
own stem cells(autologous) are either thecells from patient's ownbody or his or her cord
blood. For autologoustransplants physicians nowusually collect stem cellsfrom the peripheral bloodrather than the marrow
This procedure is easier,unlike a bone marrowharvest, it can take placeoutside of an operatingroom and the patient doesnot have to be undergeneral anaesthesia.DR.T.V.RAO MD 35
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Sources of stem cellsfrom another donor(allogeneic) are primarilyrelatives (familial-
allogeneic) or completelyunrelated donors(unrelated-allogeneic).The stem cells in thissituation are extractedfrom either the donor'sbody or cord blood
ALLOGENEIC STEM CELLS
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In this stem cells fromdifferent species aretransplanted, e.g.striatal porcine fetal
ventral mesencephalic(FVM) xenotransplantsfor Parkinson'sdisease. This has nomajor ethical concerns
and a large amount oftissue is available,however life longimmunosupression andrisk of rejection are themajor limitations
XENOGENEIC - STEM CELLS
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HOW DOES CELL THERAPY WORK?
Bone marrow transplants are an example of celltherapy in which the stem cells in a donor's marroware used to replace the blood cells of the victims ofleukemia.
Cell therapy is also being used in experiments to graftnew skincells to treat serious burn victims, and togrow new corneas for the sight-impaired.
In all of these uses, the goal is for the healthy cellsto become integrated into the body and begin tofunction like the patient's own cells.
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WHAT DISEASES CAN BECURED BY STEM CELL THERAPIES
Any disease inwhich there is
tissuedegeneration canbe a potential
candidate for stemcell therapies
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MAJOR PROGRESS IN SEVERAL IMPORTANTHEALTH PROBLEMS
Alzheimers disease
Parkinsons disease
Spinal cord injury
Heart disease
Severe burns
Diabetes
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ALZHEIMERS DISEASE AND CAN STEM
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ALZHEIMERS DISEASE AND CAN STEM
CELLS HELP?
Stem cells could,however, be geneticallymodified so as to deliversubstances to theAlzheimer brain, to stop
cells from dying andstimulate the function ofexisting cells. A recentclinical trial (Phase I) hasshown this approach tobe of some benefit topatients with Alzheimersdisease, by slowing downthe progression of the
disease. DR.T.V.RAO MD 41
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DRUG TESTING
Stem cells could allow scientists totest new drugs using human cell line
which could speed up new drug
development.Only drugs that were safe and had
beneficial effects in cell line testing
would graduate to whole animal or
human testing.
It would allow quicker and safer
development of new drugs.DR.T.V.RAO MD 42
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MAJOR TYPES OF STEM CELLS
The two broad types of
mammalian stem cells are:
embryonic stem cells thatare isolated from the inner
cell mass of blastocysts, andadult stem cells that arefound in adult tissues. In a
developing embryo, stem
cells can differentiate into allof the specialized embryonic
tissues. I
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STEM CELLS ACT AS PROGENITOR CELLS
In adult organisms, stem
cells and progenitorcells act as a repair
system for the body,replenishing specialized
cells, but also maintain
the normal turnover of
regenerative organs,such as blood, skin or
intestinal tissues.
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HISTORY OF ANIMAL CLONINGSince then, animals including mice (1998),
cows (1998), pigs (2000), cats (2001), andrabbits (2002) were successfully cloned.
RABBIT
MOUSE
CAT
Cattle
PIG
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Dolly (sheep) 1 live birth out of 29 clonedembryos
3%
Cloned mice 31 live births out of 2468 clonedembryos
1%
Cloned pigs 5 live births out of 335 clonedembryos 1%
Cloned goats 3 live births out of 85 clonedembryos
3%
Cloned cattle30 live births out of 496 cloned
embryos 6%Cloned cat 1 live birth out of 87 cloned
embryos1%
Cloned rabbits 6 live births out of 371 of cloned
embryos
1%
HOW SUCCESSFUL WAS ANIMALCLONING? VERY LOW (~1-3%)
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FIRST SUCCESS OF HUMAN EMBRYOCLONING
On February 12, 2004, SouthKorean scientists, Dr. Woo SukHwang and Dr. Shin Young Moon ofSeoul National University, reported thesuccessful creation of30 cloned
human embryos developed to theblastocyst stage and then destroyedby stem cell extraction,yielding one
embryonic stem cell line.DR.T.V.RAO MD 47
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Tens of thousands offrozen embryos areroutinely destroyed whencouples finish theirtreatment.
These surplus embryoscan be used to producestem cells.
Regenerative medicalresearch aims to develop
these cells into new,healthy tissue to healsevere illnesses.
SOURCE OF STEM CELLS FOR MEDICALTHERAPIES
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STEM CELL RESEARCH WORLDWIDE
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ADULT MU TIPOTENT STEM CELLS
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ADULT MULTIPOTENT STEM CELLS
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Adult Stem Cells
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AUTOLOGOUS STEM CELLS
While most blood stem cells
reside in the bone marrow, a
small number are present in
the bloodstream. These
Multipotent peripheral bloodstem cells, or PBSCs, can be
used just like bone marrow
stem cells to treat leukaemia,
other cancers and variousblood disorders
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multipoten
t
Speculation
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TREATMENTS BECOMES SPECIFIC
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APPLICATIONS OF STEM CELLS
Cell Replacement Therapies Cells could be stimulated to develop into specialized
cells that represent renewable sources of cells and
tissue for transplantation.
Cell replacement therapy could treat injuries andvarious genetic and degenerative conditions includingmuscular dystrophies, retinal degeneration, Alzheimerdisease, Parkinson's disease, arthritis, diabetes, spinalcord injuries, and blood disorders such as hemophilia.
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UNDERSTANDING CELL
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UNDERSTANDING CELLSPECIALIZATION
Studying human pluripotent stem cells can lead tothe identification of factors responsible fordifferentiation of stem cells into specialized cell
types. these factors may ultimately be used to drive tissue
regeneration and repair if administered therapeutically.
This work will provide basic knowledge on celldetermination and differentiation, humandevelopment, genomic imprinting and somatic cellaging.
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DEVELOPMENT AND TESTING OF
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DEVELOPMENT AND TESTING OFDRUGS
Researchers couldstudy the beneficialand toxic effects of
new medications onhuman pluripotentstem cells that havebeen developed to
mimic the diseaseprocesses.
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CAN SEX MAKE DIFFERENCE IN STEM
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CAN SEX MAKE DIFFERENCE IN STEM
CELL THERAPY ? Are there sex-specific
differences in the biology ofstem cells? (short-long term
How do sex-specificdifferences play out in termsof self-renewal anddifferentiation? (mid-longterm)
Is there existing evidencethat the sex of stem cellsaffects success of thetransplant?
XX vs.
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STEM CELLS BLINDNESS
In clinical trials atMoorfields EyeHospital in London,
surgeons restored eyesight for six patientswho lost their sightafter chemical
accidents and geneticdiseases. The patientswent under successfulstem-cell transplant.
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LIMBAL STEM CELL THERAPY
The treatment is known aslimbal stem cell therapy, andthe patients who received thetreatment suffered fromchemical burn or geneticdisease know as aniridia
By replacing the limbalstem cells, the cornea beginsto clear up as the cells are
replaced with the healthytransparent layer again.
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Research in stem cells hasopened up new horizons in thearea of treatment of disorderssuch as stroke, epilepsy, neuro-degeneration and trauma.Current research is aimed atfinding the appropriate source ofstem cells for a given indication,ways of expanding andperpetuating these cells inculture, best route ofadministration of these cells andmethods to overcome rejection
CURRENT POSSIBLE USES
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POSSIBLE USES OF STEM CELL TECHNOLOGY
Replaceable
tissues/organs
Repair of defective cell
types
Delivery of genetic
therapies
Deliverychemotherapeutic agents
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FUTURE MAKING CELLS AND
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FUTUREMAKING CELLS ANDREPLACING THE DISEASED CELLS ?
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DR.T.V.RAO MD
STEM CELL DEBATE
Some people oppose stem cell research becausethey believe that the 4-5 day old ball of cells is aliving human being. What some do not understand is
that unused embryos are trashed regardless. Manypeople, religious and non-religious, believe it is betterto use these embryos for research on how to curehuman diseases rather than to just trash them without
purpose. The underlying issue is the personal beliefwhen life actually begins.
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OBSTACLES OF STEM CELL RESEARCH
How to find the right type of stem cells?
How to put the stem cells into the right place?
Will the stem cells perform the desiredfunction in the body?
Differentiation protocols for many cell types
have not been developed.
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EMBRYONIC STEM CELLS ARE UNSTABLE ANDMUTATE IN CULTURE
Like ordinary cells,
stem cells
accumulate
significant numbers
ofmutationsover
time, including
several that could
cause them to
become tumors.DR.T.V.RAO MD
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Harvesting ES cellsdestroys the blast cyst
This is murder
ES cell researchrequires human cells
Could create acommercial market forhuman cells
This devalues life
ETHICAL DEBATE
ReproducedbypermissionofDaveCatrowandCopleyNews
Service
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DESTROYING LIFE TO CURE SOME
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If stem cells have suchpotential to relieve
suffering, why are so many
people so upset about their
use? The reason is that themost powerful type of stem
cell embryonic stem(ES) cells can only beobtained from human
embryos. Many people
think that it's wrong to
create and destroy human
embryos to treat disease
DESTROYING LIFE TO CURE SOMEONEETHICAL ?
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RELIGIOUS DEBATE OVER HARVESTING
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RELIGIOUS DEBATE OVER HARVESTINGEMBRYONIC STEM CELLS
The pro-life group generallybelieves that:
Personhood happens at, or shortlyafter, conception.
Thus, they consider the removal ofstem cells from an embryo -- aprocedure which kills the stem cells -- to be a form of murder of a humanbeing.
They argue that no potential healthbenefits to even hundreds of millionsof people can justify the murder ofother humans.
Day 5-6Blastocyst
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RELIGIOUS DEBATE OVER HARVESTING
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RELIGIOUS DEBATE OVER HARVESTINGEMBRYONIC STEM CELLS
The pro-choice group generallybelieves that:
Personhood is attained much later inpregnancy, perhaps when the fetal brain
develops consciousness during the thirdtrimester.
Thus, extracting stem cells from an fiveor ten-day old pre-embryo is notmurder.
Killing a pre-embryo, which is onlya potential human being, is
justified if it has the potential tocure diseases and extend the livesof people.
Day 5-6Blastocyst
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WHY WE SHOULD SUPPORT
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WHY WE SHOULD SUPPORTCAN HELP SEVERAL DISABLED
Human embryonic
stem cell (HESC)
research offers great
promise of cures forotherwise incurable
conditions: spinal cord
injuries, ALS,Alzheimers,
Parkinsons, etc.
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Arguments for and againsthuman cloning research.
Should we ban human
cloning? Why investors are
moving away from humancloning and why human
cloning now looks a last-
century way to fight
disease. Why some people
want to clone themselves
or even to clone the dead.
SHALL BE CLONE HUMANS ?
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FUTURE APPLICATIONS
Stem Cells may one day help scientists to regeneratecells lost in diseases like:
Repair heart muscle after a heart attack
Pancreas cells lost in diabetes
Neurons lost in Alzheimers
Retinal cells causing blindness
Understand the cell growths of cancers Help organ transplantation
DR.T.V.RAO MD 75
S C O S C S S OG SS G
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8/4/2019 Stem Cells and Progress of Science
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RESEARCH ON STEM CELLS IS PROGRESSING INSPITE OF SEVERAL RESTRICTIONS
DR.T.V.RAO MD 76
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8/4/2019 Stem Cells and Progress of Science
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ARE WE CROSSING OUR LIMITS IN SCIENCE ???
DR.T.V.RAO MD 77
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8/4/2019 Stem Cells and Progress of Science
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Programme created by Dr.T.V.Rao MD from
several web based and documented information
for Medical and Paramedical Professionals inthe Developing World
Email