CELL DIVISION AND STEM CELLS

Cells divide by mitosis for growth and repair.They divide by meiosis to

produce gametes for sexual reproduction. Stem cells differentiate into
specialised cells during the development of organisms

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 Each human body cell contains 46 chromosomes. These can
be arranged into 23 pairs. Each chromosome in a pair carries
the same types of genes.
 Cell division
 Cells divide when:
 an organism grows
 organisms need to replace worn-out cells
 organisms need to repair damaged tissue
 During these processes, the cell undergoes a type of cell
division called mitosis.

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 In mitosis, two cells called daughter cells are produced. It is
essential that any new daughter cells produced contain genetic
information that is identical to the mother cell, and that the number
of chromosomes remains constant.

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 Two parents are needed in sexual reproduction. During this process
the nucleus of a male gamete fuses with the nucleus of a female
gamete, producing a new cell called a zygote .
 In human beings, each gamete contains 23 chromosomes, half the
number found in the other cells of the body. When the male and
female gametes fuse, they become a zygote, which in turn becomes a
new embryo containing the full 46 chromosomes – half from the
father and half from the mother.
 This fusion of gametes is called fertilisation.

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STEM CELLS DEFINITION AND
PROPERTIES.

 Stem cells are generally defined as clonogenic cells capable of

both cell renewal and multilineage differentiation since they are
thought to be undifferentiated cells with varying degree of potency
and plasticity.

 PROPERTIES OF STEM CELLS.

1. STEM CELLS CAN DIVIDE AND RENEW ITSELF.
 Stem cells have a special ability to divide and renew themselves
for extended periods of time. In fact an initial population of stem
cells can produce millions of cells within few month in a laboratory
setting.
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 When the produced cells remain unspecialized for a longer
duration, it indicates the long term self renewal capacity of cells.
2. STEM CELLS ARE UNSPECIALIZED.
The unspecialized nature of stem cells is an important one. It means
that stem cells lack the specific parts that allow them to performed
specialized functions in the body. A stem cells does not have a
specialized function but it has the capacity to differentiate into a
specialized cells that can carry out these functions.

3. STEM CELLS CAN GIVE RISE TO SPECIALIZED CELLS.

The ability of stem cells to give rise to specialised cells is a crucial
one. in this process of differentiation, unspecialised stem cells
produced specialised cells. It is thought that a cell’s genes regulate the
internal signal that trigger this process. This gene carry the specific
code, or instructions, for all parts and function of a cell.

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CLASSIFICATION OF STEM CELLS
There are different type of stem cells.
They can be classified on the basis of
1. Extent to which they can differentiate.

i. Totipotent cells
ii. Pluripotent cells
iii. Multipotent cells
iv. Oligopotent cells
v. Unipotent cells

2. On the basis of origin

i. Embryonic germ cells 10
ii. Umbilical cord stem cells.
iii. Embryonic stem cells.
iv. Adult stem cells.

3. On the basis of their source.

i. Autologous stem cells.
ii. Allogenic stem cells.
iii. Xenogenic stem cells

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ON THE BASIS OF EXTENT TO WHICH THEY
CAN DIFFERENTIATE.
1. Totipotent cells: These cells have the ability to form an entire
organism .
It has potential to generate all the cells and tissues that make up an
embryo and that support its development in utero. In human this ability
is preserve untill 8 cell stage.
Cells produced by first few division of fertilised egg are totipotent.
These cells can differentiate into embryonic and extra embryonic cells
type.
These are most versatile of stem cells type.
It has potential to give rise to any and all human cells, such as brain,
liver, blood, or heart cells etc .
After four days of embryonic cell division, the cells begin to specialize
into pluripotent stem cells.
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PLEURIPOTENT CELLS
 These cells can give rise to any cell type found in primary germinal
layer of embryo, i.e. ectoderm, mesoderm and endoderm.
 These are the cells isolated from inner cell mass at blastocyst-stage
of embryo.
 These are the descents of totipotent cells and can differentiate into
cells derive from three germ layer . They can give rise to all tissue
type, however cannot give rise to an entire organism.
 There are four classes of pleuripotent cells
1. Embryonic stem cells.
2. Embryonic germ cells.
3. Embryonic carcinoma cells .
4. Multipotent adult progenitor cells.
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MULTIPOTENT STEM CELLS
 These cells can give rise to all cell types within a particular lineage.
Multipotent stem cells are found in adult tissues; perhaps most organs
in the body (e.g brain, liver) contain where they can replace dead or
damaged cells. These adult stem cells may also be cells that when one
accumulates sufficient mutations produce a clone of cancer cells.
 These are less plastic and more differentiated stem cells. They give
rise to limited range of cells within a tissue type. They can become
one of the several types of cells within a given organ.
 e.g blood stem cells can develop into red blood cells, white blood
cells, or platelets.

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 OLIGOPOTENT STEM CELLS
 These cells have limited differentiation potential.
e g stem cells from adult. Lymphoid/myeloid organs.

 Unipotent stem cells These cells can differentiate into only a particular defined
lineage. They are found in adult tissue and in comparison with other stem cells
have the lowest differentiation potential.
 They also have therapeutic potential to treat injuries and disease.

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ON THE BASIS OF ORIGIN OF STEM CELLS

 Stem cells are classified as of embryonic and and post natal

derivation based on developmental stage from which they are
obtained.
 Stem cells of embryonic origin:
 Embryonic stem cells can be obtained from an embryo at the
blastocyst stage.
 They have finite replicative life span after which they can no longer
divide and one said to have reached a proliferative senescence.
 Embryonic stem cell sources.
 Embryos created via I V F.
 Embryos or fetuses obtained through elective abortion .
 Embryos created via SCNT (somatic cell nuclear transfer, or
cloning) 17
 When these cells in culture are grown under certain conditions, they
remain undifferentiated, but if they are allowed to clump together to
form embryoid bodies, they begin to differentiate spontaneously and
can be directed to form specific cells by altering the chemical
composition of the culture medium. Embryonic germ cells are
collected from foetal tissue from gonadal ridge.

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 Stem cells of fetal origin :
 Haematopoietic stem cells are present in umbilical cord blood.
 Multipotent and pluripotent stem cells can be obtained from fetus
after termination of pregnancy. The only safe and relatively feasible
source of fetal stem cells is fetal blood.

 Umbilical cord stem cells :

 In the umbilical cord, we can find two types of stem cells source, i.e
the umbilical cord epithelium, derived from the amniotic membrane
epithelium and the umbilical cord blood.

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 Stem cells of adult origin:
 Progenitor and multipotent stem cells are present in adults. Somatic
stem cells can generate for example liver, panceras, bone, cartilage,
neural system cells. Haematopoietic stem cells are present in the
blood, originating from bone marrow after stimulation. An adult
stem cell is a multipotent stem cells in adult humans that is used to
replace cells that have died or lost function.

 Bone marrow transplants are carried out:

in cases of blood cell cancer, eg leukaemia and lymphoma. when
blood cells have been destroyed, e g during cancer treatment.

 They can be obtained from bone marrow, muscle, synovium, skin, 21

decidous teeth, nerve tissue etc.
 Adult stem cells can be found in limited numbers in
several regions of the body, eg:
 brain
 eyes
 blood
 heart
 liver
 bone marrow
 skin
 muscle
 Adult stem cells can differentiate into related cell
types only, for instance, bone marrow cells can
differentiate into blood cells and cells of the immune
system, but not other cell types. 22
ON THE BASIS OF SOURCE
Autologous Stem Cells
Sources of the patient's own stem cells (autologous) are either the cells
from patient's own body or his or her cord blood. For autologous
transplants physicians now usually collect stem cells from the
peripheral blood rather than the marrow .This procedure is easier,
unlike a bone marrow harvest, it can take place outside of an operating
room and the patient does not have to be under general anaesthesia.

Allogeneic Stem Cells

Sources of stem cells from another donor (allogeneic) are
primarily relatives (familial- allogeneic) or completely
unrelated donors (unrelated-allogeneic). The stem cells in this
situation are extracted from either the donor's body or cord
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blood
 Xenogenic Stem Cells
 In this stem cells from different species are transplanted, e.g. striatal
porcine fetal ventral mesencephalic (FVM) xenotransplants for
Parkinson's disease. This has no major ethical concerns and a large
amount of tissue is available, however life long immunosuppression
and risk of rejection are the major limitations.
 Progenitor stem cells:
 .Stem cells act as Progenitor cells .In adult organisms, stem cells and
progenitor cells 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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HOW DOES STEM CELL THERAPY
WORK?
 Stem cells can be used to generate healthy and functioning
specialized cells, which can then replace diseased or dysfunctional
cells. It is similar to the process of organ transplantation only the
treatment consists of transplanting cells instead of organs.

 Bone marrow transplants are an example of cell therapy in

which the stem cells in a donor's marrow are used to
replace the blood cells of the victims of leukemia.

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 Cell therapy is also being used in experiments to graft new
skin cells to treat serious burn victims, and to grow new
corneas for the sight-impaired. In all of these uses, the goal
is for the healthy cells to become integrated into the body
and begin to function like the patient's own cells.

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WHAT DISEASES CAN BE CURED BY STEM
CELL THERAPIES.
 Any disease in which there is tissue degeneration can be a potential
candidate for stem cell therapies
 Major Progress in Several Important Health problems
 Alzheimer’s disease
 Parkinson’s disease
 Spinal cord injury
 Heart disease
 Severe burns
 Diabetes

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 Drug Testing
 Stem cells could allow scientists to test 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.

 Current possible uses

 Research in stem cells has opened up new horizons in the area of
treatment of disorders such as stroke, epilepsy, neuro- degeneration
and trauma. Current research is aimed at finding the appropriate
source of stem cells for a given indication, ways of expanding and
perpetuating these cells in culture, best route of administration of
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these cells and methods to overcome rejection.
 Benefits and risks associated with the use of
stem cells in medicine
 Stem cells have great potential, eg in treating
patients with currently untreatable conditions,
growing organs for transplants, and research. The
benefits of using your own stem cells include:
 no rejection
 no need to find a donor
 no need for tissue typing
 But there are clinical, ethical and social issues
associated with their use. These issues will be
different for the growth and transplant of adult,
embryonic, or therapeutically-cloned stem cells. They
will also depend on whether the stem cells are to be
used for therapy or research. 31
 Clinical issues
 There is no guarantee how successful these therapies will be, for
example, the use of stem cells in replacing nerve cells lost in
Parkinson’s disease patients.

 The current difficulty in finding suitable stem cell donors.

 The difficulty in obtaining and storing a patient’s embryonic stem

cells. These would have to be collected before birth – some clinics
offer to store blood from the umbilical cord when a person is born.

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 Mutations have been observed in stem cells cultured for a number of
generations, and some mutated stem cells have been observed to
behave like cancer cells.

 Cultured stem cells could be contaminated with viruses which would

be transferred to a patient.

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 ASCs are also known as postnatal stem cells or somatic stem cells
which can be obtained from bone marrow, umbilical cord, pancreas,
adipose tissue, and dental pulp.
 Dental stem cell is a type of adult mesenchymal cell which has ease
of access and is feasible source of stem cells. There are various
sources of dental stem cells in the orofacial region such as dental
pulp stem cells, dental follicle stem cells, stem cells of apical papilla,
periodontal ligament stem cells, tooth germ progenitor cells, and
immature dental pulp stem cells.
 There are numerous applications of this progenitor cell such as
continued root formation, regeneration of an immature tooth with
extensive pulp damage, periodontal regeneration, biological tooth,
and stem cell-based therapies, which are gaining worldwide
attention because of its copious benefits.

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 The advances in applications of dental stem cells seem to be
exceeding in the near future, for which specialized skills and
knowledge in this arena are of prime significance. Hence, there is a
need to acquire more knowledge about dental stem cells to obtain
maximum benefits from it in the coming years. Dental stem cells in
India are still at the budding stage, and there seems to be limited
awareness regarding dental stem cells.

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STEM CELLS IN DENTISTRY .
 The severe bone resorption in edentulous areas makes it difficult to
restore the missing teeth with dental implants or denture treatment.
 Therefore, stem cell and tissue engineering therapies are expected to
provide a novel capability to regenerate large defects in periodontal
tissues and alveolar bone and to ultimately replace the lost tooth
itself.
 The tissues and organs targeted for such regenerative medicine
strategies in dentistry include the salivary gland, tongue and
craniofacial skeletal muscles, as well as the condylar cartilage of the
temporomandibular joint.

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 Dental stem cells are present in various dental tissues , including
both deciduous and permanent teeth .(periodontal ligament, Apical
papilla, and dental follicle)

 Constructing complex structures like periodontium, which

provides functional correlation between tooth or implant
and surrounding jaw, could effectively improve modern
dentistry.
 Recent studies have demonstrated that they can also be
used to generate dentin and alveolar bone, and to
bioengineer small, anatomically correct, whole tooth crowns
consisting of enamel, dentin and pulp tissue.

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SOURCES OF STEM CELLS IN DENTISTRY

1. Dental pulp stem cells - DPSC’s

2. Stem cells from Human Exfoliated Deciduous teeth – SHED.
3. Human cementum derived cells- HCDC’s
4. Stem cells from root apical papilla- SCAP
5. Periodontal ligament stem cells- PDLSC’s
6. Epithelial stem cells from the labial cervical loop of rodent incisor
7. Epithelial stem cells from developing molars- EpSCs
8. Stem cells from the dental follicle – DFSC

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 Steps involved in regeneration of tooth are:

1. Harvesting of adult stem cells.

2. Seeding the stem cells into the scaffold that provides optimized the
environment.
3. Cells are instructed with targeted soluble molecular signals spatially
.
4. Confirming the gene expression profile of the cells for the next
stage in odontogenesis .

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 PERIODONTAL REGENERATION
 Cell-based therapeutics for periodontal tissue regeneration may be
engineered
 The use of adult stem cells and of reprogrammed stem cells by
lineage switching or direct conversion may represent a promising
future of periodontal therapeutics .
 Periodontal stem cells are common progenitors of fibroblasts,
cementoblasts, and osteoblasts that form the tissues of the
periodontium & contribute to periodontal regeneration.

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 Stem cell mediated root regeneration offers opportunities to
regenerate bio-root and its associated periodontal tissues, which are
necessary for maintaining the physiological function of teeth. It can
be used for the regeneration of teeth in patients without adequate
bone support.

 Isolation of human periodontal ligament stem cells.

 Dental pulp and periodontal ligament were obtained from
normal human impacted third molars

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 Formation of cementum and regeneration of root . Cultures of
primary human cementum derived cells (HCDC’s) have been
established from healthy teeth using a collagenase .

 With these cells discrete colonies containing cells with fibroblast

like morphology are formed and when these colonies become
sufficiently large, cells from individual colonies were isolated and
cultured.

 The presence of cementoblast progenitors in cultures of

bovine dental follicle cells has been reported and their
differentiation capacity has been demonstrated.

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 DENTAL PULP STEM CELLS(DPSCS):

 DPSCs were the first type of dental stem cells to be isolated which
were obtained by enzymatic digestion of the pulp tissue. These are
fibroblast- like in morphology, clonogenic, in nature and can
maintain their high proliferation rate even after extensive
subculturing
 Historically, dental stem cells were first isolated by Gronthos and
co- workers from the dental pulp and exfoliated deciduous teeth.
Dental stem cells can also be extracted from the apical papilla of
shed primary teeth. They are rich source of mesenchymal stem cells.

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 They are thought to originate from the cranial neural crest which
explains their multi- differentiation potential with the capacity to
give rise to various cell lineages such as adipocytes, osteocytes,
chondrocytes, and myocytes: odontoblasts, and neuronal cells.
DPSCs improve angiogenesis and play a role in cardiac repair
Animal studies regenerative potential of DPSCs and clinical studies
also characterized their role in bone augmentation in tooth extraction
sockets.

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 Human adult dental pulp stem cells(ADPSCs) and stem cells from
human exfoliated deciduous teeth(SHEDs) are self-renewing stem
cells residing within the perivascular niche/ cell rich zone of the
dental pulp. ADPSCs and SHEDs can be obtained without any
complications from impacted adult wisdom teeth and naturally
exfoliated deciduous respectively Culturing ADPSCs with various
differentiation media demonstrated their dentinogenic osteogenic,
adipogenic, neurogenic, chondrogenic and myogenic differentiation
capabilities.

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DENTAL STEM CELLS EXTRACTION
 Stem cells are taken from deciduous teeth ( presence of vital pulp
and physiological resorption of the root no bigger than two third),
third molars ( age limit upto 28 years), extracted teeth for
orthodontic reason , mesiodens or trauma.
 Method of isolation of dental pulp stem cells:
 l). After extraction keep the tooth in saline/ fresh milk in a container
with frozen gel packs for delivery to the laboratory
 2). Then, it is transferred to the tooth bank into a vial containing
hypotonic phosphate buffered saline solution(to prevent the tissue
from drying during transport). The vial is then carefully sealed and
placed into a thermette, which is then placed into an insulated metal
transport vessel

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.
 3). At the tooth bank a stringent protocol is followed for cleaning the
tooth surface by various disinfectants.
 4). The pulp tissue is isolated from the pulp chamber and the cells
are then cultured in a mesenchymal stem cell(MSC) medium.
Different cell lines(odontogenic, adipogenic, and neural) can be
obtained by making various changes in MSC medium.

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 Cultivation of Dental stem cells
 Stem cells suspension (DPSCs) are cultivated in a special media for
human mesenchymal progenitor cells the laboratory dishes holding
the primary culture contain a so called cell+ surface. Finally, they
are treated with trypsin –EDTA and divided into laboratory dishes
with standard surfaces.
 There are two sources from which dental pulp elements are
developed
 Dental mesenchyme from neural crest
 vascular mesenchyme
 The pulp stem cells division time is as follow:
 12-50 hours for the first forty division in cell population, while afer
50 cell divisions, the division time increases to 60-90 hours 55
 Drawbacks of using dental pulp stem cells:
 I.) Stem cells may have a low survival rate.
 2.) Transport and isolation of stem cells is critical.
 3.) Challenges methodology and clinical translation.
 4.) Chances of contamination are high.
 5.) Trained expertises are required.
 6.) Cost is way to high.

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 Dental Pulp Stem Cell Mechanoresponsiveness:
Effects of Mechanical Stimuli on Dental
Pulp Stem Cell Behavior.
 Marrelli M1, Codispoti B1, Shelton RM2, Scheven
BA2, Cooper PR2, Tatullo M1, Paduano F1

 Dental pulp stem cells in chitosan/gelatin scaffolds

for enhanced orofacial bone regeneration.
 Bakopoulou A, Georgopoulou Α, Grivas I, Bekiari C,
Prymak O, Loza Κ, Epple M, Papadopoulos GC, Koidis
P, Chatzinikolaidou Μ.
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VASCULARIZED BONE GRAFTS
 Vascularized bone grafts can be used for reconstruction of bone in
recipient site that is morbid due to infection, radiation injury or
extensive damage to local blood vessels.
 They are generated by placing BMSC’s in collagen sponges, which
are then wrapped around an artery and vein. These cells are
subsequently wrapped with Teflon to prevent collateral ingrowth of
blood vessels.
 After several weeks, these bony rudiments are perfused entirely by
artery and vein which they surround, and then they can be moved to
another site where blood vessels can be reattached to existing blood
vessels in the margins of the recipient site

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 Anastomosis occurred by day 11, with most BMSCs associating
closely with the network. Although initially it was immature and
highly permeable, at 4 weeks the network was mature., Initiation of
scaffold mineralization also occurred by this period.
 Some human-derived vessels were still present at 5 months, but the
majority of the graft vasculature became functionally remodeled
with host cells.
 So clinically relevant progenitor sources for pericytes and
endothelial cells can serve to generate highly functional
microvascular networks for tissue engineered bone graft

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 .Dental pulp of human exfolliated deciduous teeth contains
multipotent stem cells from human exfoliated deciduous teeth
(SHED). Were identified to be a population of highly proliferative,
clonogenic cells capable of differentiating into a variety of cell types
including neural cells, adipocytes, and odontoblasts. Thus, exfoliated
teeth may be an unexpected unique resource for stem-cell therapies
including autologous stem-cell transplantation and tissue
engineering.

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 stem cells of apical papilla. A unique population of dental stem cells
known as stem cells from the root apical papilla (SCAP) is located at
the tips of growing tooth roots . The apical papilla tissue is only
present during root development before the tooth erupts into the oral
cavity . SCAP have the capacity to differentiate into odontoblasts
and adipocytes .These cells are CD24+ but expression is down
regulated upon odontogenic differentiation in vitro coincident with
alkaline phosphatase up regulation

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 Dental follicle progenitor cells:
The dental follicle is a loose ectomesenchyme-derived connective
tissue sac surrounding the enamel organ and the dental papilla of the
developing tooth germ before eruption . It is believed to contain
progenitors for cementoblasts, PDL and osteoblasts. Dental follicle
cells (DFC)form the PDL by differentiating into PDL fibroblasts that
secrete collagen and interact with fibres on the surfaces of adjacent
bone and cementum.

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 Bone marrow-derived cells (BMDCs)
 They have the potential to engraft into several tissues after injury,
but whether they can become dental tissue-specific progenitor cells
under normal conditions and the relationship of these cells to the
tissue-resident cells are unknown. bone marrow progenitor cells
communicate with dental tissues and become tissue-specific
mesenchymal progenitor cells to maintain tissue homeostasis.

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 GROWTH FACTORS DEFINITION: Growth factors are
polypeptides which have the ability to bind to specific receptors on
the target cells (in this case the stem cells and other pulpal cells) and
modulate or facilitate certain activities like migration, proliferation,
differentiation, and apoptosis

 DIFFERENT TYPES
 Bone morphogenetic proteins (BMPs)
 Platelet derived growth factor (PDGF) • Transforming growth factor
(TGF) • Fibroblast growth factors (FGF) •
 Insulin like growth factors (IGF) •
 Nerve growth factor (NGF)
 Vascular endothelial growth factor (VEGF) FGF TGF BMPs 65
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 SCAFFOLDS
 All multicellular living organisms have natural
scaffolds that surround the cells and provide
structural support for formation and maintenance of
tissues and organs.
 Natural scaffolds
 Synthetic scaffolds
 Collagen Platelet rich plasma
 Fibrin Glycosaminoglycans Polylactic acid (PLA),
Polyglycolic acid (PGA),
 Polycaprolactone (PCL).
 Polyethyleneglycol (PEG) HYDROXYAPETITE
COLLAGEN 3- D HYDROGEL 67
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 DPSCs has the potential for regenerating both pulp and dentin
tissues. Thus, may offer an alternative method to save teeth that may
have compromised structural integrity. The possibility of pulp tissue
regeneration is restricted by several factors, like, the dental pulp has
minimal collateral blood supply, impairing the ability of the immune
system to combat infection.

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CONCLUSION

 As the research in this field continuing on with

promising future , we may be able – in the near
future – to implant a biotooth primordium into a
patient’s gum in the place of a removed orlost
tooth. Although many studies have to be
conductedbefore applying such therapeutic
modalities,the dental stem cells represent a
powerful toolwhich holds a significant potential
for advancementin the field of regenerative
dentistry and medicine

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