P.Anusha M.

Pharmacy 119A2S0302

What are stem cells

Stem cells are the raw material from which all of the bodys mature, differentiated cells are made. Stem cells give rise to brain cells, nerve cells, heart cells, pancreatic cells, etc. They have the potential to replace cell tissue that has been damaged or destroyed by severe illnesses.
They can replicate themselves over and over for a very long time.

Two Kinds of Stem Cells

Embryonic (also called pluripotent)

stem cells are capable of developing into all the cell types of the body. Adult stem cells are more difficult to identify, isolate, and purify.

Stem cells based upon their source

Embryonic stem cells obtained from the

undifferentiated inner mass cells of a blastocyst, (an embryo that is between 50 to 150 cells) Umbilical cord blood derived from the blood of the placenta and umbilical cord after birth Adult stem cells undifferentiated cells found among differentiated cells of a specific tissue and are mostly multipotent cells Bone marrow Adipose tissue CNS stem cells Umbilical Cord Blood

Embryonic Stem Cells:

Two Sources of Embryonic Stem Cells Excess fertilized eggs from IVF (in-vitro fertilization) clinics

Therapeutic cloning (somatic cell nuclear transfer)

Stem cell therapy

Stem cell therapy is most often regarded as a means to provide

replacement of specific cells for repair of a damaged or failing organ. Therefore, great efforts have been made to identify both ideal sources of cells, and those with the greatest differentiate potential, so that multiple cellular phenotypes can be obtained.

However, recent

studies have provided compelling evidence that, in some disease states, engraftment of transplanted stem cells may not, in fact, be required to gain a therapeutic benefit.

 stem cells exert their benefit by selectively seeking out

areas of injury/damage within the body and serving as a reservoir of cytokines and growth factors that promote repair of the endogenous cellular pool within the damaged organs. These findings suggest that stem cells would be ideally suited as vehicles for delivering therapeutic genes or drugs to damaged/ diseased tissues.

Adult stem cell gene manipulation

The prototypic example of adult stem cells, the hematopoietic stem cell. Although they are relatively rare in the human body, these cells can be readily isolated from bone marrow or after mobilization into peripheral blood. After in vitro manipulation, these cells may be retransplanted into patients by injection into the bloodstream, where they travel automatically to the place in the bone marrow in which they are functionally active. Another adult bone marrow-derived stem cell type with potential use as a vehicle for gene transfer is the mesenchymal stem cell, which has the ability to form cartilage, bone, adipose (fat) tissue. The traditional method to introduce a therapeutic gene into hematopoietic stem cells from bone marrow or peripheral blood involves the use of a vector derived from a certain class of virus, called a retrovirus. However, these particular retroviral vectors were only capable of transferring the therapeutic gene into actively dividing cells. Since most adult stem cells divide at a relatively slow rate, efficiency was rather low. Vectors derived from other types of retroviruses (lentiviruses) and adenoviruses have the potential to overcome this limitation, since they also target non-dividing cells. The major drawback of these methods is insertional mutagenesis. and difficult to ex-vivo manipulation.

embryonic stem cell gene manipulation

Following derivation, human embryonic stem cells are easily accessible for controlled and specific genetic manipulation. When this facility is combined with their rapid growth, remarkable stability, and ability to mature in vitro into multiple cell types of the body, human embryonic stem cells are attractive potential tools for gene therapy. First, human embryonic stem cells could be genetically manipulated to introduce the therapeutic gene. This gene may either be active or awaiting later activation, once the modified embryonic stem cell has differentiated into the desired cell type.

 The therapeutic gene needs to be introduced into the

cell type used for therapy. Genes may be introduced into cells by transfection or transduction. Transfection utilizes chemical or physical methods to introduce new genes into cells. Transduction utilizes viral vectors for DNA transfer. Viruses, by nature, introduce DNA or RNA into cells very efficiently.

 parameter that must be carefully monitored is the random integration

into the host genome, since this process can induce mutations or serious gene dysfunction. However, several copies of the therapeutic gene may also be integrated into the genome, helping to bypass positional effects and gene silencing. Positional effects are caused by certain areas within the genome and directly influence the activity of the introduced gene. Gene silencing refers to the phenomenon whereby over time, most artificially introduced active genes are turned off by the host cell, a mechanism that is not currently well understood

 Gene silencing and positional effects can be avoided by

to introduce the gene of interest specifically into a defined region of the genome by the gene targeting technique ( homologous recombination ).

Gene manipulated stem cell delivery

Direct delivery
Therapeutic gene Genetically modified ES

Cell based delivery

embryonic stem cells

Therapeutic gene

The therapeutic gene is packaged into a delivery vehicle such as a retrovirus

Adult stem cells are isolated and propagated in the laboratory. Adult stem cells

Invitro differentiated stem cell

The therapeutic g is packaged into a delivery vehicle su as a retrovirus and introduced into th cells.

and injected into the patient

The genetically modified cells are reintroduced into the patient

Applications of manipulated stem cells for delivery as systemic therapeutics

Any disease in which there is tissue degeneration can be a

potential candidate for stem cell therapies. Alzheimers disease Parkinsons disease Spinal cord injury Heart disease Severe burns Diabetes

Stem cells for Alzheimers disease

Stem cells could, however, be genetically modified so as to deliver substances to the Alzheimer brain, to stop cells from dying and stimulate the function of existing cells.

Manipulated stem cells for Parkinson's disease

Stem cells for diabetes

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.

Limbal stem Cell therapy

The treatment is known as limbal stem cell therapy, and the patients who received the treatment suffered from chemical burn or genetic disease know as aniridia By replacing the limbal stem cells, the cornea begins to clear up as the cells are replaced with the healthy transparent layer again.

References

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