Chapter 1 : Introduction & Aim of the work

Stem Cells

:Under supervision
Prof.Dr: Mohammed Shabaan Hemeda
Lecturer of forensic medicine
portsaid university
:BY
Somaya Mohammed Attia
ID:
3114
 Acknowledgement

- First and foremost, all praises and thanks to Allah, the Almighty, for his assistance and favor to
accomplish this work.

- I would like to express my deepest appreciation to my, Prof.Dr: Mohammed Shabaan Hemeda
Lecturer of forensic medicine at Faculty of Medicine - Port Said University who guided me
throughout my project and offered with constructive criticism that made my project what it is.

- Nevertheless, I express my gratitude toward my colleagues for their kind co-operation and
encouragement, which help me in completion of this study.

I
 Chapter 1 : Introduction & Aim of the work

Index
1 List of figures

2 List of abbreviations

Chapter 1 : Introduction 3
Introduction &
3 Aim of the work 3
Aim of the work
4 Chapter 2 : Historical perspectives
literature Review
Stem cells’ definition 4
Challenges in stem cell research 5
Classification of stem cells on the basis of potency 8
(Totipotent – Pluripotent – Multipotent – Oligopotent -
Unipotent)
Classification of stem cells on the basis of their source 9
(Embryonic stem cells - Adult stem cells - Pluripotent stem
cells).
Cancer stem cells in drug discovery 11
Stem cells as an alternative for arthroplasty 13
Stem cells complement animal models in drug discovery 16
Stem cell culture 20
Target identification and evaluation tools aiding stem cell- 20
based drug discovery.
Applications of stem cells in treatment of certain diseases 21
(Skin replacement - Brain cell transplantation - Treatment for
diabetes)
5 Chapter 3 Conclusion 26
6 Chapter 4 References

II
 List of figures
Ll

Page
No. Title
Number

1 Characters of stem cell: replication and differentiation 6

Illustrates Stem cells can turn into any type of cell

2 7
before they become differentiated.

3 Classification of stem cell on basis of potency 9

4 Sources of adult stem cells 11

Targeted screens for drug development with immediate

5 19
clinical applicability.

6 Stem cell culture and stem cell lines 22

Examples of applications of stem cells in treatment of

7
diseases

8 Applications of stem cells to wounds

9 Types of differentiations of Multipotent Stem Cells.

III
Chapter 1 : Introduction & Aim of the work

List of abbreviations

Abbreviation Full Form

iPS cells Induced pluripotent stem cells

ONFH Osteonecrosis of the femoral hip

MSCs Mesenchymal stem cells:

OA osteoarthritis

THA total hip arthroplasty

hESCs Human embryonic stem cells

GM-CSF granulocyte colony stimulating factor

TNF-α tumor necrosis factor

IFNγ interferon gamma

IV
Chapter 1 : Introduction & Aim of the work

Chapter 1 : Introduction & Aim of the work

 Section I: Introduction:
Stem cells are defined as cells that have clono-genic and self-renewing
capabilities and differentiate into multiple cell lineages. Stem cells are found in all
of us, from the early stages of human development to the end of life. Stem cells are
basic cells of all multicellular organisms having the potency to differentiate into
wide range of adult cells. Self-renewal and totipotency are characteristic of stem
cell. Though totipotency is shown by very early embryonic stem cells, the adult
stem cells possess multipotency and differential plasticity which can be exploited
for future generation of therapeutic options. [1]
All stem cells may prove useful for medical research, but each of the
different types has both promise and limitations. For decades, researchers have
been studying the biology of stem cells to figure out how development works and
to find new ways of treating health problems. The scientific researchers and
medical doctors of today hope to make the legendary concept of regeneration into
reality by developing therapies to restore lost, damaged, or aging cells and tissues
in the human body.[1]

 Section II: Aim of the work:

Increasing awareness and knowledge about stem cells and what have
reached in executing their applications for therapeutic and beneficial purposes, and
realizing the recent limits established in this particular field.

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Chapter 4 : References

Chapter 2 : literature Review

 Section I: Historical perspectives

The history of stem cell research had a benign, embryonic beginning in the
mid 1800's with the discovery that some cells could generate other cells. In the
early 1900's the first real stem cells were discovered when it was found that some
cells generate blood cells. The term "stem cell" was proposed for scientific use by
the Russian histologist Alexander Maksimov in 1908. Bone marrow transplant
between two siblings successfully treated SCID in 1968. Haemopoietic stem cells
were discovered in human cord blood in 1978. [2]
James Thomson and coworkers derived the first human embryonic stem cell
line at the University of Wisconsin–Madison in 1998. More recently, in 2005,
scientists at Kingston University in England were purported to have found another
category of stem cell. These were named cord blood embryonic-like stem cells,
which originate in umbilical cord blood. Korean researcher Hwang Woo-Suk
(2004–2005) claimed to have created several human embryonic stem cell lines
from unfertilized human oocytes.[2]
The first published study of successful cartilage regeneration in the human knee
using autologous adult mesenchymal stem cells is published by clinicians from
Regenerative Sciences in 2008.
Embryonic stem cell isolated from a single human hair was reported in 2008.
[3]

Over the last few years, national policies and debate amongst the public as well
as religious groups, government officials and scientists have led to various laws
and procedures regarding stem cell harvesting, development and treatment for

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Chapter 4 : References

research or disease purposes. The goals of such policies are to safeguard the public
from unethical stem cell research and use while still supporting new advancements
in the field. [3]

 Section II: Stem cells’ definition

A stem cell is a non-specialized, generic cell which can make exact copies of
itself indefinitely and can differentiate and produce specialized cells for the various
tissues of the body. Stem cells are cells found in most, if not all, multi-cellular
organisms. They are characterized by self-renewal and potency as shown in figure
1 i.e. - the ability to renew themselves through mitotic cell division and
differentiating into a diverse range of specialized cell types. They are vital to the
development, growth, maintenance, and repair of our brains, bones, muscles,
nerves, blood, skin, and other organs.[4]

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Chapter 4 : References

Figure (1): Characters of stem cell: replication and differentiation

 Section III: Challenges in stem cell research

Stem cells need much more study before their use can be expanded.
Scientists must first learn more about how embryonic stem cells develop. This will
help them understand how to control the type of cells created from them. Another
challenge is that the embryonic stem cells available today are likely to be rejected
by the body. And some people find it morally troubling to use stem cells that come
from embryos. [28]
Scientists also face challenges when using adult pluripotent stem cells.
These cells are hard to grow in a lab, so researchers are looking into ways to
improve the process. These cells are also found in small amounts in the body.
There is a greater chance that they could contain DNA problems. Clinical trials
that use stem cell therapies are currently being done in the U.S. If you are
interested in trying this therapy to treat a certain condition, ask your healthcare
provider how to find out about trials available in your area. [28]

 Section IV: Adult stem cells

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Chapter 4 : References

Figure (2) illustrates Stem cells can turn into any type of cell before they become
differentiated.
A person’s body contains stem cells throughout their life. The body can use
these stem cells whenever it needs them. Also called tissue-specific or somatic
stem cells, adult stem cells exist throughout the body from the time an embryo
develops. The cells are in a non-specific state, but they are more specialized than
embryonic stem cells. They remain in this state until the body needs them for a
specific purpose, say, as skin or muscle cells. [30]

Day-to-day living means the body is constantly renewing its tissues. In some
parts of the body, such as the gut and bone marrow, stem cells regularly divide to
produce new body tissues for maintenance and repair. [30]

Stem cells are present inside different types of tissue. Scientists have found
stem cells in tissues, including:

 the brain

 bone marrow

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Chapter 4 : References

 blood and blood vessels

 skeletal muscles

 skin

 the liver

However, stem cells can be difficult to find. They can stay non-dividing and
non-specific for years until the body summons them to repair or grow new tissue.
Adult stem cells can divide or self-renew indefinitely. This means they can
generate various cell types from the originating organ or even regenerate the
original organ entirely. This division and regeneration are how a skin wound heals,
or how an organ such as the liver, for example, can repair itself after damage. [30]

In the past, scientists believed adult stem cells could only differentiate based
on their tissue of origin. However, some evidence now suggests that they can
differentiate to become other cell types, as well.[30]

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Chapter 4 : References

 Section V: Classification of stem cells on the basis of potency

Stem cells can be classified by the extent to which they can differentiate into
different cell types. These four main classifications are totipotent, pluripotent,
multipotent, or unipotent as shown in figure (3).

Figure (3): Classification of stem cell on basis of potency

Totipotent:
The ability to differentiate into all possible cell types. Examples are the
zygote formed at egg fertilization and the first few cells that result from the
division of the zygote.[6]
Pluripotent:
The ability to differentiate into almost all cell types. Examples include
embryonic stem cells and cells that are derived from the mesoderm,
endoderm, and ectoderm germ layers that are formed in the beginning stages
of embryonic stem cell differentiation.[6]

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Chapter 4 : References

Multipotent:
The ability to differentiate into a closely related family of cells. Examples
include hematopoietic (adult) stem cells that can become red and white
blood cells or platelets.[6]
Oligopotent:
The ability to differentiate into a few cells. Examples include (adult)
lymphoid or myeloid stem cells. [6]
Unipotent:
The ability to only produce cells of their own type but have the property of
self-renewal required to be labeled a stem cell. Examples include (adult)
muscle stem cells.

 Section VI: Classification of stem cells on the basis of their sources

The easiest way to categorize stem cells is by dividing them into two types:
Early or embryonic and mature or adult. Early stem cells, often called embryonic
stem cells, are found in the inner cell mass of a blastocyst after approximately five
days of development. Mature stem cells are found in specific mature body tissues as
well as the umbilical cord and placenta after birth.[6]

Embryonic stem cells:

Embryonic stem cells are self-replicating pluripotent cells that are
potentially immortal. They are derived from embryos at a developmental stage
before the time of implantation would normally occur in the uterus. The embryos

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Chapter 4 : References

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. [6]
Adult stem cells:
Adult stem cells are undifferentiated totipotent or multipotent cells, found
throughout the body after embryonic development, as shown in figure (4), that
multiply by cell division to replenish dying cells and regenerate damaged tissues.
The primary role of adult stem cells in a living organism is to maintain and repair
the tissue in which they are found. Unlike embryonic stem cells, which are defined
by their origin (the inner cell mass of the blastocyst), the origin of adult stem cells
in some mature tissues is still under investigation. [6]

Pluripotent stem cells:

A third type of stem cell, with properties similar to embryonic stem cells,
has emerged. Scientists have engineered these induced pluripotent stem cells (iPS
cells) by manipulating the expression of certain genes - 'reprogramming' somatic
cells back to a pluripotent state.[6]
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Chapter 4 : References

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Chapter 4 : References

 Section VII: Cancer stem cells in drug discovery

Cancerous cells also exploit the same properties of self-renewal and

multipotency that make stem cells so attractive in regenerative medicine and drug
discovery. Recent studies have identified a small unique population of cells known
as cancer stem cells, or tumor-inducing cells, that reside in tumors. These cells
arise from oncogenic transformation of either stem cells or progenitor cells. Cancer
stem cells or tumor-inducing cells have “stem-like” character, a slow proliferation
rate, a high capacity for self-renewal, resistance to standard chemical/radiation
therapy, and a propensity to differentiate into actively proliferating tumor cells. [29]

Cancer stem cells have been isolated from many tumor types, including
brain, renal, colon, and prostate tumors, as well as hematopoietic cancers, and
therapeutic strategies are being developed to target cancer stem cells for apoptosis
or cell cycle arrest specifically and thereby eradicate tumors more effectively than
current treatments. Cancer drug discovery programs have, to a large extent,
employed immortalized cell lines or primary tumor tissue for in vitro assay. This
approach has been largely unsatisfactory in developing effective therapeutics for
cancer.[29]

Cancer drug discovery programs involving cancer stem cells as a platform

offer a discovery process with a high degree of therapeutic efficiency. This is
particularly evident from the drug discovery process in leukemia. The fact that
cancer stem cells feed the cancer growth and promote resistance to existing
chemotherapeutic drugs makes them candidate cells for drug discovery screens. In
cancer of the hematopoietic system, cancer stem cells have been well characterized

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Chapter 4 : References

in chronic myelogenous leukemia, acute myelogenous leukemia, and acute

lymphoblastic leukemia. Drug discovery programs targeting CD33 in acute
myelogenous leukemia and the ABL kinase inhibitor, imatinib mesylate, in chronic
myelogenous leukemia have been very effective. Biomarkers such as CD33 have
been shown to be uniquely characteristic of the cancer stem cells of acute
myelogenous leukemia, and efforts to design immunotherapy against such
biomarkers are becoming very useful. [29]

From the perspective of drug discovery and translational research, cancer

stem cells provide a solid platform with significant implications for designing
therapeutic approaches and developing long-term treatment. It has to be noted that
in vitro propagation of cancer stem cells does not mimic factors governing the
microenvironment of cancer stem cells, such as hypoxia, but they do offer a
platform for evaluating drug screens with a higher degree of confidence, thereby
facilitating preclinical trials and clinical trials of antitumor therapies. [29]

Research reports indicating that cancer stem cells can be selectively targeted
without harming normal stem cells offer a possibility to perform targeted drug
discovery screens against cancer stem cells from different tumors. [29]
 Section VIII: Stem cells as an alternative for arthroplasty:

One of the biggest fears of professional sportsmen is getting an injury, which

most often signifies the end of their professional career. This applies especially to
tendon injuries, which, due to current treatment options focusing either on
conservative or surgical treatment, often do not provide acceptable outcomes. Stem
cell therapy is a method that can help delay or avoid arthroplasty when treated in

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Chapter 4 : References

the early stages of osteoarthritis, when cartilage has not yet been completely
destroyed. [32]
Mesenchymal stem cells are used to restore joints. The essence of the
technique is to introduce them into the damaged area. Problems with the tendons
start with their regeneration capabilities. Instead of functionally regenerating after
an injury, tendons merely heal by forming scar tissues that lack the functionality of
healthy tissues. Factors that may cause this failed healing response include
hypervascularization, deposition of calcific materials, pain, or swelling.
Additionally, in addition to problems with tendons, there is a high probability of
acquiring a pathological condition of joints called osteoarthritis (OA). OA is
common due to the avascular nature of articular cartilage and its low regenerative
capabilities. [32]
Although arthroplasty is currently a common procedure in treating OA, it is
not ideal for younger patients because they can outlive the implant and will require
several surgical procedures in the future. These are situations where stem cell
therapy can help by stopping the onset of OA. However, these procedures are not
well developed, and the long-term maintenance of hyaline cartilage requires further
research. [32]
Osteonecrosis of the femoral hip (ONFH) is a refractory disease associated
with the collapse of the femoral head and risk of hip arthroplasty in younger 20
populations. Although total hip arthroplasty (THA) is clinically successful, it is not
ideal for young patients, mostly due to the limited lifetime of the prosthesis. An
increasing number of clinical studies have evaluated the therapeutic effect of stem
cells on ONFH. Most of the authors demonstrated positive outcomes, with reduced
pain, improved function, or avoidance of THA. [32]

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Chapter 4 : References

Currently, scientists are considering two mechanisms of action of

mesenchymal stem cells:
 MSCs differentiate (turn) into cells of a certain type and replace damaged
or missing cells in bones, cartilage, ligaments or tendons.
 MSCs secrete paracrine factors, specific molecules that reduce
inflammation, which plays a key role in cartilage degeneration.

In general, treatment contributes to the restoration of damaged articular

structures. In the first few days after administration, the inflammatory process
decreases until it is completely eliminated, which is accompanied by pain relief. In
addition, cell therapy contributes to:
 Strengthening and growth of new blood vessels, normalization of blood
circulation (articular tissues receive more nutrients and growth factors
that stimulate the healing process of damaged areas of bones, cartilage,
tendons and ligaments).
 Slowing down the progression of the disease, which makes it possible to
delay or avoid endoprosthetic surgery.
 Restoration of mobility and stability of the joint, so that the patient can
return to a full life. [32]

In recent years, numerous clinical trials have been conducted that have
proven the effectiveness of the treatment of joint diseases with stem cells. As an
example, there are several studies conducted at the Prodromos Stem Cell Institute
in Maryland. [32]
1) Treatment with autologous (taken from the patient) mesenchymal stem
cells has proven effective in the treatment of osteoarthritis of the knee.

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Chapter 4 : References

Regenerative therapy has shown good results in relieving the symptoms

of the disease for at least 6 months after the procedure.
2) Cellular therapy using MSCs derived from adipose tissue and bone
marrow, in combination with the use of platelet-rich plasma, is an
effective alternative to knee arthroplasty in patients with moderate
arthrosis. In some cases, after treatment, patients did not even need to
take medication.
3) Injections of autologous mesenchymal cells for the treatment of osteo-
and inflammatory arthritis are recognized as completely safe. During the
study, scientists did not identify adverse events specific to stem cells,
such as infection, oncogenesis, or chondrolysis. [32]

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Chapter 4 : References

 Section IX: Stem cells complement animal models in drug

discovery:

Stem cells have already received enormous attention for their potential in
refining drug screens. Stem cell screens are a more cost-effective strategy than
animal testing. They are also seen as a process of refining, reducing, and possibly
replacing animal testing procedures. However, animal models offer a whole system
setup for testing the effects and side effects of drugs. It has to be noted that the
response to drugs in animal models may not be the same as in humans. Animal
models differ from those in humans in a number of ways. In particular, they do not
reflect the ethnic diversity of humans and their response to drugs may be different
to that seen in humans. In addition, animal models are costly and time-consuming.
[29]

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Chapter 4 : References

Stem cells offer an alternate platform to overcome the limitations of ethnic

diversity, provide a uniform response to a drug which can correlate with the human
response, and are cost- and time-effective. Unfortunately, the current status of stem
cell research does not allow us to substitute for whole animal testing, as seen with
animal models. However, stem cells are very promising substitutes in terms of
single organ toxicity. Embryonic or adult tissue-specific stem cells that can
ultimately be grown into an entire human organ will be a gold standard for single
organ drug testing. [29]

Cell-based in vitro assays provide a scaffold to perform high throughput or high

content screens (Figure 5), but they do not reflect the complex in vivo scenario. [29]

Figure (5): Targeted screens for drug development with immediate clinical applicability.
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Chapter 4 : References

Cell-based screens do not take into consideration the cross-talk between

organs (or different cell types), or the general metabolism and side effects seen in
vivo. Hence in vitro assays have to be complemented with various in vivo assays
using different animal models. Currently, various invertebrate and vertebrate
models are available for screening the toxicity and efficacy of lead molecules. [29]

Invertebrate models of Drosophila and Caenorhabditis elegans offer a

powerful model suitable for drug discovery research. Their small size, short
generation time, relatively low cost of housing and maintenance, highly conserved
molecular pathways, and availability of various genetic and biochemical tools have
made it possible to include them in multistep drug screening processes. The ease
with which transgenics, overexpression, and mutation of proteins can be done, as
well as the relatively simple genetic cascade involved have enabled their use in
performing screens directed against Alzheimer’s disease, Huntington’s disease,
oncogenic transformation, stem cell niche, metastases, neurodegeneration,
apoptosis, and behavioral analyses. [29]

These simple models offer a setup whereby a particular process such as

neurodegeneration can be quickly screened. For example,
a Drosophila Huntington’s disease model and transgenic line provide an ideal in
vivo system for not only examining mutant Huntington gene-mediated cellular
defects, such as impairment of axonal transport, but also facilitate rapid assays for
screening and validating potential treatment to alleviate the observed cellular
defects. The relatively simple metabolic and genetic cascades and the long
evolutionary separation from humans are major disadvantages limiting the use of
these models in drug development research. [29]

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Chapter 4 : References

Higher vertebrate models such as rabbits, dogs, monkeys, and rodents have
been extensively used in drug safety testing. Among them, rodents are a very good
complement to stem cell-based screening. The availability of knockings,
conditional knockouts, and transgenic models forms a powerful support system to
evaluate the in vivo response of various lead molecules identified in stem cell-
based in vitro screens. [29]

The identification of a tissue-specific stem cell niche in rodent models offers

the advantage of developing screens directed towards manipulation of the stem cell
microenvironment to aid in understanding and developing therapeutic strategies for
various diseases, such as neurodegenerative disorders, stroke, organ transplant,
brain trauma, wound healing, and cancer. Such screens have identified molecules
that affect the progenitor pool size of the adult neural stem cell
population. However, the traditional methods of analysis in rodent models are slow
and rely extensively on analyses of tissues collected from sacrificed animals. [29]

In recent years, teleost vertebrate models, in particular zebrafish and

medaka, have become popular models for studying various aspects of
developmental biology and genetics. Their rapid external development,
transparency of embryos, husbandry, and large sample size are some of the
advantages readily offered by these models. The possibility of performing toxicity
and efficacy screening of chemicals, pharmaceuticals, and pesticides that can be
correlated in terms of human health risks are propelling this model as a choice for
toxicological or pharmacological screens. [29]

Small molecule screening to identify and characterize a molecule that

produces specific effects against various disease processes in humans has been

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Chapter 4 : References

successfully developed. The zebrafish (Danio rerio) has been more extensively
used than the medaka (Oriyzas latipes), but both show a high degree of anatomical
and physiological homology with that of other higher order vertebrates, and also
have very similar cellular structure, signaling processes, and cognitive
behavior. The collection of various mutant lines with defects in the development
and function of the various metabolic processes, and the availability of various
biochemical, molecular, and genetic techniques has facilitated the development of
various in vivo drug screens targeting development, metabolism, and physiological
conditions in terms of various human disease. [29]

 Section X: Stem cell culture

Growing cells in the laboratory is known as cell culture. Human embryonic
stem cells (hESCs) are generated by transferring cells from a preimplantation stage
embryo into a plastic laboratory culture dish that contains a nutrient broth known
as culture medium. The cells divide and spread over the surface of the dish.
However, if the plated cells survive, divide and multiply enough to crowd the dish,
they are removed gently and plated into several fresh culture dishes. The process of
replating or subculturing the cells is repeated many times and for many months.
Each cycle of subculturing the cells is referred to as a passage. Once the cell line is
established, the original cells yield millions of embryonic stem cells. Embryonic
stem cells that have proliferated in cell culture for six or more months without
differentiating, are pluripotent, and appear genetically normal are referred to as an
embryonic stem cell
line. At any stage in
the process, batches

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Chapter 4 : References

of cells can be frozen and shipped to other laboratories for further culture and
experimentation. Stages are shown in figure (6).

• Section XI: Target identification and evaluation tools aiding

Figure (6): Stem cell culture and stem cell lines stem cell-based
drug discovery

Drug discovery screens using stem cells are a new and immensely necessary
resource. The most commonly used screening process involves growing cells as a
monolayer culture and exposing them to libraries of new chemical entities. The
final readout in most of the high throughput scenarios is based on imaging.
Imaging-based screens are quick and can be automated with ease. Time-lapse
assays can also be performed to understand the onset of the phenotype. Imaging-
based readouts can simply employ bright field images to understand the impact of
the compound on colony size, morphology, proliferation, and cell number. These
image-based readouts can also be combined with immunofluorescence to test the
expression/inhibition of markers of interest. Commercially available assays, which
use fluorescent readouts to measure cell proliferation, apoptosis, and toxicity, can
be employed with relative ease in a high throughput screen scenario. [29]

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Chapter 4 : References

Other methodologies, such as quantitative polymerase chain reaction and

fluorescence-activated cell sorting, can be performed at a low to medium
throughput screening level and offer wide endpoint assays, such as measuring cell
cycle and proliferation, as well as quantifying expression of biomarkers at the
RNA or protein level. Techniques such as microarrays, chip-sequencing, single cell
illumina-based sequencing, and proteomic approaches, eg, affinity column
purification and matrix-assisted laser desorption ionization time-of-flight mass
spectrometry, can be used effectively as a high-end analysis for target
identification and characterization of the lead compound. [29]

 Section XII: Applications of stem cells

The goal of any stem cell therapy is to repair a damaged tissue that can't heal
itself. Ongoing research on stem cell therapies gives hope to patients who would
normally not receive treatment to cure their disease but just to alleviate the
symptoms of their chronic illness. Stem cell therapies involve more than simply
transplanting cells into the body and directing them to grow new, healthy tissue. It
may also be possible to coax stem cells already in the body to work overtime and
produce new tissue. There are some good applications of stem cells in treatment of
certain diseases as stem cells have good characteristics as shown in figure (7). [8]

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Figure (7): Examples of applications of stem cells in treatment of diseases

A) Stem cells in treatment of wounds:

Wound healing is a complex process that involves mitosis, inflammation,
angiogenesis, synthesis, and remodeling of the extracellular matrix. When wound
healing does not occur, the wound may become chronic and need additional
interventions. [8] Research has been performed on the effects of using MSCs in the
treatment of wounds, both with indirect and direct delivery to the wound site as
shown in figure (8). New studies have shown that MSCs are home to sites of injury
and provide therapeutic impact. Several studies have suggested that MSCs home to
regions of injury by specific trafficking to chemokine ligand 7 (CCL7). [10, 11]
Once the MSCs reach the point of injury, the MSCs exit the vasculature in
the connective tissue stromal region. [9] The MSCs respond to the specific tissue
milieu while at the same time contributing to the milieu through the secretion of
biomolecules. [13] It has shown that MSCs can be activated using cytokines such as
granulocyte colony stimulating factor (GM-CSF), tumor necrosis factor (TNF-α),
or interferon gamma (IFNγ) to enhance activity and therapeutic impact. [12, 15]

Figure (8) Application of stem cells to wounds

B) Stem
cells in
orthopedics:
It is known that MSCs have the capability to move chemotactically to areas
of inflammation and infection in an organism’s tissue. MSCs actively contribute to

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Chapter 4 : References

tissue regeneration such as intravenous routes, secreting soluble factors, and

regulating inflammatory responses. [16]
Further, MSCs also secrete factors which promote bone regeneration. In
cartilage regeneration, MSCs have been used as a therapeutic to repair damage. In
research conducted by Shafiee and colleagues, MSC based cartilage repair in rabbit
models that had full thickness cartilage defects showed promise with improved
healing, measured through macroscopic scores. At six months after the study, the
MSCs showed effectiveness in chondrocyte transplantation, as well as tissue
regeneration. [17]
The largest problems are big bone defects, often as a result of infection,
tumors, trauma, insufficient blood supply or as a post-infection consequence. [18]
These defects pose a great problem, as bone supply is greatly limited, thus creating
difficulties in producing autologous bone grafts. [18] These bone grafts also result in
high levels of morbidity in donors, as well as a heightened risk of disease
transmission or rejection in recipients. Thus, the use of MSCs as an alternative
treatment in the area of bone defects is appealing. [19]
So, stem cells can differentiate into multiple types of tissue as shown in figure 9. [19]

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Chapter 4 : References

Figure (9): Types of differentiations of Multipotent Stem Cells.

C)
Stem cells in hematological pathology:
Hematopoietic stem cells (HSCs) are often used as a treatment in
hematological pathologies but can cause many adverse reactions, such as bleeding,
graft versus host disease (GvHD), and other forms of rejection. [20] MSCs have the
potential to successfully aid in HSC engraftment and prevent rejection with their
immune-suppressive properties. MSCs also generate cytokines that aid
hematopoiesis and could enhance the efficacy of MSCs in bone marrow recovery
after chemotherapy and/or radiation. [20]

D) Stem cells for inherited and neurological disease:

MSCs have demonstrated their ability to change into neurons and astrocytes.
Due to these observations, mouse models have been used to test MSC transplants
on mice with acid sphingomyelinase, a neurodegenerative disease. Infusion of the
MSCs resulted in a delay in the start of neurological abnormalities and improved
overall survival in the mouse model. Based on this experiment, a study was started
to determine the effectiveness of MSC transplantation into human patients with
amyotrophic lateral sclerosis, a disease that causes degeneration of motor neurons
and muscle functionality. [20]
Central nervous system injury (CNS) situations can be caused by a stroke,
trauma, or an underlying neurological condition. In CNS, neural MSCs (NSCs) and
MSCs are used for regeneration purposes to create new cells to replace those that
were lost. However, this process has not been completely effective due to oxidative
stress and toxic by-products, which can affect MSC transplantation. [21]

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Chapter 4 : References

This finding causes slowing of tissue regeneration, as well as reduced

longevity. In yet another neurological disease, Parkinson’s, MSCs have been
shown effective at inhibiting inflammatory cytokine production, a main factor that
contributes to the disease. Scientists from the University Hospital of Tubingen in
Germany observed yet another effective way to deliver MSCs to neurological
patients, through the nose. Studies were performed in a Parkinson’s induced rat
model, with intranasal administration of bone marrow MSCs. In these rodents,
MSCs were found in the olfactory bulb, cortex, striatum, cerebellum, brain stem,
hippocampus, and spinal cord up to 4.5 months after administration, providing data
that suggested MSCs could proliferate in vivo successfully. [21]
It was observed that intranasal administration increased tyrosine hydroxylase
levels in the lesioned ipsilateral striatum and substantia nigra, while decreasing
levels of the toxin 6- hydroxydopamine. [22, 23]

E) Stem cells in cystic fibrosis:

Cystic fibrosis (CF) is a genetically inherited disease which results in
mutations in the cystic fibrosis transmembrane regulator (CFTR) gene. The
mutation in this disease impacts almost every organ of the body, but the major
cause of morbidity and mortality is the inability to control lung infection and
inflammation. Since bone marrow MSCs have both anti-inflammatory and
antimicrobial properties studies were done to investigate the potential of using
MSCs as a therapeutic in the murine model of CF lung infection and inflammation.
[24]

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Chapter 4 : References

In this model CF mice lose considerable weight without resolution and often
succumb to the infection. Therapeutic bone marrow MSCs in this model resulted in
weight changes similar to control mice with improved gross lung pathology and
decreased cellular recruitment into the lung. Further, the bone marrow MSCs
shifted the pulmonary differential from predominantly neutrophils to a more
evenly distributed differential of both neutrophils and macrophages. [24, 25, 26]

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Chapter 4 : References

Chapter 3 : Conclusion

Stem cell therapies give hope to patients who would normally not receive
treatment to cure their disease. Stem cells have a bright future for the therapeutic
world by promising stem cell therapy. Hoping to see new horizon of therapeutics
in the form of bone marrow transplant, skin replacement, organ development, and
replacement of lost issues such as hairs, tooth, retina and cochlear cell

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 Chapter 4 : References

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