Therapy, introduction, applications

2. OVERVIEW OF STEM CELLS
 Stem cell biology
 Requirements & Potency definition
 Types of stem cell
 Stem cell collection
 Cryopreservation of stem cell
 Stem cell banking
 Stem cell therapy-disease treatment
 Benefits& limitation of stem cell
 Application of stem cell

3. STEM CELL BIOLOGY
 The science of stem cells is known as stem cell biology. It has been one of the
fascinating areas of biology today provides cell-based therapies to treat
reparative diseases.
STEM CELLS
 The undifferentiated animal cells that are capable of proliferation and
differentiation to do specialized functions are called stem cells.
1. Actual stem cells
2. Potential stem cells
 The body is made up of about 200 different kinds of specialised cells such as
muscle cells, nerve cells, fat cells and skin cells.
 All cells in the body come from stem cells (Stem cells are the building blocks
of the blood and immune systems.
 Stem cells can self-renew to make more stem cells
 differentiate into a specialized cell type
WHY ARE STEM CELLS SPECIAL?

4. CHARACTERISTICS OF STEM CELLS
 Stem cells are cells are capable of continuous proliferation for several months.
Sub-culturing the stem cells for more than six months and observing the culture
under a microscope for a clump formation. The stem cells do not form clumps.
 They have identical chromosomes as in diploid cells of the adult organisms.
 They can be sub-cultured, stored in liquid nitrogen.
 They are pluripotent. This is confirmed by allowing the ESCs.
 They have no tumour inducing property. When the ESCs are injected into
immunosuppressed mouse, they do not induce tumours in that mouse.
POTENCY DEFINITIONS
 Pluripotent: Stem cells that can differentiate into any tissue type except for
placenta tissue.
 Multipotent: Stem cells that can differentiate into multiple cells in a closely
related family of cells.

5. MULTPOTENT PLURIPOTENT
blood stem cell
found in
bone marrow
MULTIPOTENT
only specialized types of blood cell:
red blood cells, white blood cells,
platelets
differenti
ation
Stem cells that can become many
types of cells in the body are called
pluripotent
Stem cells that can become only a few
types of cells are called Multipotent
Tissue stem cells
(Multipotent)
Embryonic stem
cells (pluripotent)

6. TYPES OF STEM CELLS
1. EMBRYONIC STEM CELLS
2. ADULT STEM CELLS
EMBRYONIC STEM CELLS
 The undifferentiated cells isolated from embryos at blastocyst stage are called
(ESCs).
 These cells are capable of continuous proliferation and differentiation, leading to
the development of many different cell types in the adult. Hence, they are said to be
Pluripotent.
 ESCs are found as the inner cell mass in the blastocyst.
ESCs (Germ layer)
Endoderm Ectoderm Mesoderm
(Thymus,thyroid,lung, (Skin,nerves,head, (Bonemarrow,blood
liver,pancreas, gastro eyes, ears) cells,muscles, heart,
-intestinal tract) blood vessels)

7. EMBRYONIC STEM (ES) CELL CULTURE
feeder layer
Blastocyst
embryonic stem cells taken from
the inner cell mass
outer layer of cells
= ‘trophectoderm’
cells inside
= ‘inner cell mass’
culture in the lab
to grow more cells
fluid with
nutrients
embryonic stem
cell
PLURIPOTENT
differentiation
All possible types of specialized cells

8. ADULT STEM CELLS
 The stem cells isolated from fully developed organs are called adult stem cells (ASCs).
They can be isolated from liver,skin,heart, bonemarrow,blood stream,muscles, pancreas.
 In these organs, adult stem cells are found in discrete populations. If a part of these
organs are injured, the adult stem cells regenerate to heal up the injury. Thus they
regenerate tissues lost due to injury, tear or disease.
FEATURES OF ASCs:
 They are tissue specific stem cells. Each ASC type gives only a particular cell type
present in the organ from which it was isolated.
 Stem cell obtained from one organ may give rise to a cell type of yet other organ. This is
called plasticity.

9. STEM CELL COLLECTION
Peripheral Blood Stem Cell
Collection
Bone Marrow Harvest:
 Stem cells can be collected from the
blood. This technique does not require
surgery. It conventional bone marrow
harvest. Prior to the collection, the
donor is given a medication to promote
the growth and release of stem cells
from the bone into the blood. The stem
cells are then collected using a special
machine called a Cell Separator.. Stem
cells are generally collected using this
method here at the Leukemia.
 Stem cells can be collected directly
from the bone marrow spaces, most
often from the pelvic bones. Several
puncture sites are made along the bone
and the cells are removed using a
needle. This procedure is known as
a bone marrow harvest. Note that this
technique is used less often here at the
Leukemia.

10. CRYOPRESERVATION
 Cell lines (SC) have been selected
for long time storage at -196ºC.
 The selected cell lines are
subjected to various test such as
Cytological, biochemical,
physiological & immunological
tests.
 COS media 3 contains 100µg/ml
Streptomycin sulfate, 20% Calf
serum, 100 units/ml Penicillin-G,
1% L-Glutamine& Dulbecco’s
Modified Eagle Medium per litre.
 During preservation, cells are kept
on Solid carbon dioxide (-79ºC) or
Liquid Nitrogen (-196ºC).

11. STEM CELL (CORD BLOOD)BANKING
 An amniotic stem cell bank is a facility that stores stem cells derived from amniotic
fluid for future use. The first private amniotic stem cell bank (life cell) in the US was
opened by Biocell Center in October 2009 in Medford.
 Cord blood banking newborn's umbilical cord and placenta and storing it for future
medical use. Cord blood contains potentially lifesaving cells called stem cells. Life cell-
largest private bank 2004 in Chennai.
 Cord blood(bio-insurance) is a rich source of blood stem cells.
 Stem cells are also found in bone marrow, human embryos, fetal tissue, hair follicles, baby
teeth, fat, circulating blood, and muscle. Every part of the human body contains some
stem cells, but most are not a rich enough source to be harvested for therapeutic
applications.

12. HUMAN EMBRYONIC STEM CELLS
 The undifferentiated cells present in the inner cell mass of
human blastocyst embryo are called human embryonic stem
cells (HESCs).
 (HESCs) have been used in cell-based therapies to cure several
diseases in man. Stem cell research since 1998.
 The HESCs can be used to prepare translating cells to cure
Parkinson’s disease, diabetes, heart disease, vision loss and
hearing loss, autism, Alzheimer's diseases,etc. Cord blood stem
cells have been used successfully to treat more than 70
different diseases.
 Researches on stem cells to treat these diseases are progressing
every year towards success.

13. APPLICATIONS IN DISEASE TREATMENT
Pancreas
Pancreas
LEUKEMIA
 TYPE 1 DIABETES
 HEART DISEASE(MUSCLE REPAIR)
 PARKINSON’S DISEASE(PD)

14. LEUKEMIA
 Leukemia is cancer of the blood or bone marrow(which produce blood
cells), person suffer abnormal production of blood cells
[Leukocytes(WBC)].
FUNCTION OF THE BONEMARROW:
Marrow-large bones of adults produces blood cells,4% our total
body weight consists of bone marrow.
TWO TYPES OF BONE MARROW:
Red marrow:
WBC - Fight disease
RBC - Carry oxygen
PLATLETS - Essential for blood clotting
Yellow marrow : Inside middle section of the long bones.

15. TYPES OF LEUKEMIA:
Acute Lymphocytic Leukemia
Chronic Lymphocytic Leukemia
Acute Myelogenous Leukemia
Chronic Myelogenous Leukemia
SYMPTOMS OF LEUKEMIA:
Blood clotting is poor, affected immune system, anemia, fever, nausea, night
sweats, tiredness ,headache & weight loss.
CAUSES OF LEUKEMIA:
Artificial ionizing radiation,HIV, hair dyes, genetic predisposition(family
history).
TREATMENTS:
Chemotherapy , radiation therapy, stem cell transplant.

16. STEM CELL TRANSPLANTATION FOR LEUKEMIA:
It is also called Hematopoietic progenitor cell transplantation. S.C collect
from the peripheral blood& umbilical cord blood.
STEM CELL TRANSPLANT: (BONE MARROW)
A stem cell transplant is a procedure to replace diseased bone marrow
with healthy bone marrow. Before a stem cell transplant, receive high
doses of chemotherapy or radiation therapy to destroy diseased bone
marrow. Then receive an infusion of blood-forming stem cells that help to
rebuild bone marrow. May receive stem cells from a donor, or in some
cases may be able to use your own stem cells. A stem cell transplant is
very similar to a bone marrow transplant.
TYPES:
 Autologous stem cell transplant
 Allogeneic stem cell transplant

17. AUTOLOGOUS STEM
CELL TRANSPLANT
ALLOGENIC STEM CELL
TRANSPLANT
Stem cell collected from the
patients blood
Harvested
Frozen
Stored until needed
(Radiation & Chemotherapy)
Destory the cancer cells
Stem cell taken from matching
donor
Relative& family
Umbilical cord blood
(Sibling)
Donor’s stem cells are the right
match ( HLA test), compare the
patient’s blood & tissue type with
blood samples from the donor.

18. STEM CELL TRANSPLANTATION FOR LEUKEMIA

19. DIABETES
Diabetes mellitus (DM):
It is simply known as diabetes, is a group of metabolic diseases in
which there are high blood sugar levels over a prolonged period. The normal
cells of pancreas that produce insulin are destroyed by the patients own immune
system, leading to diabetes in humans.
TYPES OF DIABETES
There are three main types of diabetes mellitus:
Type 1 DM:
Results from the body's failure to produce enough insulin. This form was
previously referred to as "insulin-dependent diabetes mellitus" (IDDM). Is an
autoimmune disease in which the body attacks beta cells in the pancreas that
secrete the important hormone insulin.

20. Type 2 DM :
Insulin resistance, cells fail to respond to insulin properly. As
the disease progresses a lack of insulin may also develop. This form
was previously referred to as "non insulin-dependent diabetes
mellitus" (NIDDM) or "adult-onset diabetes". The primary cause is
excessive body weight and not enough exercise.(family history).
Gestational diabetes :
Is the third main form and occurs when pregnant women
without a previous history of diabetes develop a high blood glucose
level.
SYMPTOMS:
This high blood sugar produces the symptoms of frequent
urination, increased thirst, and increased hunger. Untreated, diabetes
can cause many complications. Acute complications
include coma. heart disease, stroke, kidney failure, foot
ulcers and damage to the eyes.

21. PREVENTION AND TREATMENT :
Involves a healthy diet, physical exercise, not
using tobacco and being a normal body weight. Blood
pressure control and proper foot care are also important for
people with the disease.
Type 1 -diabetes must be managed with insulin injections.
Type 2- diabetes may be treated with medications with or
without insulin. Insulin and some oral medications can
cause low blood sugar.
Gestational diabetes- usually resolves after the birth of the
baby.

22. STEM CELL BASED THERAPY
Isolated islet cells from pancreas of foetus, purified and introduced
them into pancreas of patients suffering Type-I diabetes.
It increased the insulin concentration for a few days and thereafter
the insulin concentration decreased slowly.(transplanted cells
remained in the pancreas).
The gene PDX1- insulin producing gene in cells. Isolated PDX1
gene& introduced into human embryonic stem cells to differentiate
them into insulin secreting cells (β-cells of pancreas).
Cells were introduced into pancreas using a laparoscopic device. The
introduced cells formed of islets like groups of cells and produced
insulin continuously in the patients.

23. This technique has been tried to cure diabetes in clinical laboratories

24. PARKINSON’S DISEASE (PD)
 Parkinson's disease is a progressive nervous
system disorder that affects the person moves,
including they speak and write.
 The PD is a neurodegenerative disorder
seen in 2% of human population over 65 years
of age. It is due to progressive degeneration
and loss of dopamine-producing neurons.
TREATMENT:
 Embryonic stem cell transplantation was used. Lab-differentiated ESCs into
dopamine, producing neurons for transplantation in patient suffering from PD.
 The PD cannot be treated with known drugs.
 New Technique Could Treat Parkinson’s Using Patient’s Own Brain Cells
 Scientists have successfully implanted nerve stem cells into the brain of a
monkey, targeting the area destroyed by Parkinson’s disease.

25. EMBRYONIC STEM CELL TRANSPLANTATION
Nurr 1 gene was introduced into mouse ESCs to differentiate them into
dopamine producing neurons.
Then the differentiated neurons are transplanted into the brain of the
Parkinson's model mouse.
The differentiated neurons reinnervated the brain, released dopamine
and improved the motor function.
Attempts to differentiate dopamine producing neurons from HESCs for
transplantation in Parkinson's patients.
Neurotransplantation for Parkinson's disease available in the near
future.

26. STEM CELL TREATMENT

27. STEM CELL THERAPY FOR HAIR LOSS
 Recent technology for the treatment of Alopecia (Baldness) i.e.
Male & female pattern hair loss & hair loss associated with
various disorders.
 It is a revolutionary treatment for hair fall. The stem cells
promote new hair growth within 3 to 4 weeks of treatment. The
hair density increases by 30 to 40% by three sessions of
treatment.
 On an average six sessions are required once in a week to 10
days along with which supplements are given.
 Stem cells activate the dead hair follicles and convert them into
growing healthy new hair follicles.

29. BENEFITS OF ADULT STEM CELL
RESEARCH
• Easy to obtain
 Potentially limitless in supply
 Patients can use their own stem cells for treatment and
therapy
 Adult stem cells are politically neutral
 Not offensive to any major interest group nor do they
generate controversy.

30. LIMITATIONS OF ADULT STEM CELL
 The isolation of some types of ASC, for example the isolation of
neural cells from a patient's brain, would be impractical.
 Where a person suffers from a genetic disorder or some types of
cancers, ASC isolated from that individual will retain the
damaging genetic alterations underlying the disease and so be of
little therapeutic value.
 Maintenance in culture is difficult.

31. REFERENCE
www.stemcellcentre.edu.au
www.stemcellchannel.com.au
Hiremath; M. B. and Nerli; R. B. (2008). “Advances in Stem Cell
Research”. CURRENT SCIENCE, VOL. 95, NO. 3, 10 AUGUST 2008.
Bio-Medicine (2009). “Life Cell in India can store Cord Blood Stem Cells
and Save Live”. Retrieved from [Online] Available http://www.bio-
medicine.org/medicine-news/-u2018Life-Cell-u2019-In-India-Can-store--
Cord-Blood- Stem-Cells-and-Save-Live--6033-1/
Gupta, DK et al. Stem cell therapy: Hope and scope in pediatric practice. J
of Indian Assoc Pediatric Surgery. 2005:10;138-41.
Biotechnology- V. Kumaresan.