What are Stem Cells?

Stem cells are a class of undifferentiated cells that are able to differentiate into specialized cell types.
Commonly, stem cells come from two main sources:

Embryos formed during the blastocyst phase of embryological development (embryonic stem
cells) and

Adult tissue (adult stem cells).

Adult stem cells

Adult or somatic stem cells exist throughout the body after embryonic development and are found
inside of different types of tissue. These stem cells have been found in tissues such as the brain,
bone marrow, blood, blood vessels, skeletal muscles, skin, and the liver. They remain in a quiescent
or non-dividing state for years until activated by disease or tissue injury.

Adult stem cells can divide or self-renew indefinitely, enabling them to generate a range of cell types
from the originating organ or even regenerate the entire original organ. It is generally thought that
adult stem cells are limited in their ability to differentiate based on their tissue of origin, but there is
some evidence to suggest that they can differentiate to become other cell types.

Embryonic stem cells

Embryonic stem cells are derived from a four- or five-day-old human embryo that is in the blastocyst
phase of development. The embryos are usually extras that have been created in IVF (in vitro
fertilization) clinics where several eggs are fertilized in a test tube, but only one is implanted into a
woman.

Sexual reproduction begins when a male's sperm fertilizes a female's ovum (egg) to form a single cell
called a zygote. The single zygote cell then begins a series of divisions, forming 2, 4, 8, 16 cells, etc.
After four to six days - before implantation in the uterus - this mass of cells is called a blastocyst. The

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blastocyst consists of an inner cell mass (embryoblast) and an outer cell mass (trophoblast). The
outer cell mass becomes part of the placenta, and the inner cell mass is the group of cells that will
differentiate to become all the structures of an adult organism. This latter mass is the source of
embryonic stem cells - totipotent cells (cells with total potential to develop into any cell in the body).

9-week Human Embryo from Ectopic

In a normal pregnancy, the blastocyst stage continues until implantation of the embryo in the uterus,
at which point the embryo is referred to as a fetus. This usually occurs by the end of the 10th week of
gestation after all major organs of the body have been created.

However, when extracting embryonic stem cells, the blastocyst stage signals when to isolate stem
cells by placing the "inner cell mass" of the blastocyst into a culture dish containing a nutrient-rich
broth. Lacking the necessary stimulation to differentiate, they begin to divide and replicate while
maintaining their ability to become any cell type in the human body. Eventually, these undifferentiated
cells can be stimulated to create specialized cells.

Stem cell cultures

Human embryonic stem cell colony

Stem cells are either extracted from adult tissue or from a dividing zygote in a culture dish. Once
extracted, scientists place the cells in a controlled culture that prohibits them from further specializing

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 or differentiating but usually allows them to divide and replicate. The process of growing large
numbers of embryonic stem cells has been easier than growing large numbers of adult stem cells, but
progress is being made for both cell types.

Stem cell lines

Once stem cells have been allowed to divide and propagate in a controlled culture, the collection of
healthy, dividing, and undifferentiated cells is called a stem cell line. These stem cell lines are
subsequently managed and shared among researchers. Once under control, the stem cells can be
stimulated to specialize as directed by a researcher - a process known as directed differentiation.
Embryonic stem cells are able to differentiate into more cell types than adult stem cells.

Potency

Stem cells are categorized by their potential to differentiate into other types of cells. Embryonic stem
cells are the most potent since they must become every type of cell in the body. The full classification
includes:

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.

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.

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.

Oligopotent - the ability to differentiate into a few cells. Examples include (adult) lymphoid or myeloid
stem cells.

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.
Embryonic stem cells are considered pluripotent instead of totipotent because they do not have the
ability to become part of the extra-embryonic membranes or the placenta.

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Identification of stem cells

Although there is not complete agreement among scientists of how to identify stem cells, most tests
are based on making sure that stem cells are undifferentiated and capable of self-renewal. Tests are
often conducted in the laboratory to check for these properties.

One way to identify stem cells in a lab, and the standard procedure for testing bone marrow or
hematopoietic stem cell (HSC), is by transplanting one cell to save an individual without HSCs. If the
stem cell produces new blood and immune cells, it demonstrates its potency.

Clonogenic assays (a laboratory procedure) can also be employed in vitro to test whether single cells
can differentiate and self-renew. Researchers may also inspect cells under a microscope to see if
they are healthy and undifferentiated or they may examine chromosomes.

To test whether human embryonic stem cells are pluripotent, scientists allow the cells to differentiate
spontaneously in cell culture, manipulate the cells so they will differentiate to form specific cell types,
or inject the cells into an immunosuppressed mouse to test for the formation of a teratoma (a benign
tumor containing a mixture of differentiated cells).

Research with stem cells

Scientists and researchers are interested in stem cells for several reasons. Although stem cells do not
serve any one function, many have the capacity to serve any function after they are instructed to
specialize. Every cell in the body, for example, is derived from first few stem cells formed in the early
stages of embryological development. Therefore, stem cells extracted from embryos can be induced
to become any desired cell type. This property makes stem cells powerful enough to regenerate
damaged tissue under the right conditions.

Organ and tissue regeneration

Tissue regeneration is probably the most important possible application of stem cell research.
Currently, organs must be donated and transplanted, but the demand for organs far exceeds supply.

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Stem cells could potentially be used to grow a particular type of tissue or organ if directed to
differentiate in a certain way. Stem cells that lie just beneath the skin, for example, have been used to
engineer new skin tissue that can be grafted on to burn victims.

Cardiovascular disease treatment

A team of researchers from Massachusetts General Hospital reported in PNAS Early Edition (July
2013 issue) that they were able to create blood vessels in laboratory mice using human stem cells.

The scientists extracted vascular precursor cells derived from human-induced pluripotent stem cells
from one group of adults with type 1 diabetes as well as from another group of healthy adults. They
were then implanted onto the surface of the brains of the mice.

Within two weeks of implanting the stem cells, networks of blood-perfused vessels had been formed -
they lasted for 280 days. These new blood vessels were as good as the adjacent natural ones.

The authors explained that using stem cells to repair or regenerate blood vessels could eventually
help treat human patients with cardiovascular and vascular diseases.

Brain disease treatment

Additionally, replacement cells and tissues may be used to treat brain disease such as Parkinson's
and Alzheimer's by replenishing damaged tissue, bringing back the specialized brain cells that keep
unneeded muscles from moving. Embryonic stem cells have recently been directed to differentiate
into these types of cells, and so treatments are promising.

Cell deficiency therapy

Healthy heart cells developed in a laboratory may one day be transplanted into patients with heart
disease, repopulating the heart with healthy tissue. Similarly, people with type I diabetes may receive
pancreatic cells to replace the insulin-producing cells that have been lost or destroyed by the patient's
own immune system. The only current therapy is a pancreatic transplant, and it is unlikely to occur
due to a small supply of pancreases available for transplant.

Blood disease treatments

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Adult hematopoietic stem cells found in blood and bone marrow have been used for years to treat
diseases such as leukemia, sickle cell anemia, and other immunodeficiencies. These cells are
capable of producing all blood cell types, such as red blood cells that carry oxygen to white blood
cells that fight disease. Difficulties arise in the extraction of these cells through the use of invasive
bone marrow transplants. However hematopoietic stem cells have also been found in the umbilical
cord and placenta. This has led some scientists to call for an umbilical cord blood bank to make these
powerful cells more easily obtainable and to decrease the chances of a body's rejecting therapy.

General scientific discovery

Stem cell research is also useful for learning about human development. Undifferentiated stem cells
eventually differentiate partly because a particular gene is turned on or off. Stem cell researchers may
help to clarify the role that genes play in determining what genetic traits or mutations we receive.
Cancer and other birth defects are also affected by abnormal cell division and differentiation. New
therapies for diseases may be developed if we better understand how these agents attack the human
body.

Another reason why stem cell research is being pursued is to develop new drugs. Scientists could
measure a drug's effect on healthy, normal tissue by testing the drug on tissue grown from stem cells
rather than testing the drug on human volunteers.

CELL CYCLE

The cell cycle is the process of cell replication where two daughter cells are created from one parent
cell. The cell cycle is made up of interphase & mitosis. Mitosis is the phase of actual cell splitting
(duplication), while Interphase is made up of the G0, G1, S, and G2 phases which prepare the cell for
mitosis. The S phase involves Synthesizing new DNA and doubling the amount of DNA present so
there is enough for both daughter cells. G stands for Gap as these are the gaps in time between
mitosis and the S Phase. Cells in G0 are no longer in the cell cycle. Cells in G0 are resting & no
longer actively going through the phases of the cell cycle. Some tissue can easily transition between
G0 & G1 with the right signals while other cell types are stuck in G0.

Anti-metabolites chemotherapy drugs like 5-flourouracil & sulfa drugs (antibiotic) inhibit DNA
synthesis and stall the S phase of the cell cycle. Chemotherapy Paclitaxel inhibits microtubule action
involved in Mitosis. Therefore, cells being treated with this medication are stuck in M phase.
Permanent tissues are stuck in the G0 phase and cannot undergo further mitosis. Permanent tissues
include muscle (cardiac & skeletal) and the CNS. These tissues can only undergo hypertrophy (not
hyperplasia) to deal with increased stress and form fibrous nonfunctioning scars during the healing
process.

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Stable tissues are those that primarily reside in the G0 phase, but given the right signals can reenter
the cell cycle. For example, lymphocytes are most often in a resting period, but during infection they
are triggered to divide. Liver cells are another example of a stable tissue that can undergo replication
following injury to the organ to facilitate healing.
Labile Tissues are cells which are constantly replicating with no time in the G 0 phase. These cells act
as the stem cells for tissues such as the bone marrow and skin.

Repair and Healing

The difference between the two types of repair is the cell type that will make the replacement . In
regeneration there is a replacement for the morphofunctional cells while in healing is the replacement
by fibrous tissue.

The entire repair process is controlled by growth factors that are stored in the extracellular matrix
(ECM ) . Furthermore , the process of repair needs cell proliferation , angiogenesis and extracellular
matrix deposition. The ECM is extremely important in the healing process because it supports cell
adhesion , it is the reservoir of growth factors and, in particular , is composed of fibroblasts that
synthesize collagen which is essential in healing .

There are basically three cell types related to tissue repair : labile , stable or quiescent cells and
permanent cells . These cells are in different stages of the cell cycle. Stable cells are in G0 , ie, they
are at rest and when they receive appropriate signals, they enter into the cell cycle. They are little
specialized cells such as epithelial cells . Some cells have cell cycle shortened . It is the case of labile
cells which are induced to divide more rapidly . The permanent cells are highly specialized ones such
as neuronal cells and cardiomyocytes. We can make the analogy that the more specialized a cell is ,
the less it proliferates . Thus , only permanet cells are related to healing because, in that case, the
dead cells are replaced by fibrous tissue .

Regeneration

The inflammatory process will bring new blood vessels and ECM . Growth factors present in the ECM
will induce the labile and stable cells to enter into a shorter cell cycle in order to replace by the
parenchymal cells of the same organ cells .

Healing

Having the final components of inflammation available , the body will use angiogenesis and fibroblasts
to healing. In the affected area will be a deposit of fibroblasts ( fibroplasia ) that synthesize collagen
. Then there is the remodeling where the synthesized collagen will acquire resistance to the scar .

Healing can be further classified into first and second intention . The first intention occurs when the
lesion is not extensive and without secondary infection, while the second intention occurs when the
lesion is very extensive , reaching the stroma and the wound is contaminated .

It is important to know that what will drive the type of repair that will occur will be :
1 - the type of tissue reached
2 - the type of injury
3 - the extent of injury

As in all processes of our body , there are factors that influence the healing process . These are
called local and systemic factors . As local factors we can cite foreign bodies , lesion size and location
of the injury , for example. As systemic factors we can cite nutrition, circulatory status and metabolic
state , for example.

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