Cell Therapies 1

Question 1

Has there been an explosion of stem cell based therapy?

Answer 1

Yes:
1. Mesenchymal stem cells
2. Has significantly increased year upon year
3. Animal data, such as improving of bond generation.

Question 2

Does registering a trial ensure scientific validity?

Answer 2

No:
Therefore for the patients its hard to evaluate the risks and benefits as stem cells therapies.
This is because its hard to covince the patient with scientific evidence

Question 3

What effects to Mesenchymal stromal cells have?

Answer 3

Regenerative, they can be isolated fomr the bone marrow (via gene therapy), adipose tissue, cord(blood), or amniotic fluid.
Also from iPSC cells.

Question 4

Is there any marketing approval from the USA?

Answer 4

NO

Question 4

Where does the knowledge of MSC come from?

Answer 4

Mice and rats
But these are not considered compatible with the clinical trial, compared to the animal model:
Dosing, immune compatibility and fitness of culture adapated to MSCs.

Question 4

MSC types

Answer 4

• Allogeneic marrow MSCs for GvHD
• Fit adipose MSCs for entercutaneous fistular disease (take fats out and those cells were then frozen and revived for 48 hours)
• For fistular its trying to regenerate an area of tissue where it has gone because of the disease

Question 5

MSC modes of action

LOOK AT THE CIRCLE DIAGRAM IN THE LECTURE

Answer 5

1. Paracrine effects (secreting cytokines which affect inflamation, immunomodulatory function, secretions of exosomes which contain miRNAs and proteins) This controls cell differentation and interactions
2. Differentation (bone)
3. Cell-Cell effects (Cell membrane proteins stimulate endogenous cells to differentiate)

Question 5

MSC differentation

Answer 5

Naturally differentiate into bone, fat and cartilidge or skeletal muscle.
Can fuse with other cells and transfer genetic materila with cells that are ins situ and chnages responses.

Question 5

MSC paracrine activity

Answer 5

VEGF to promot angiogenesis, to promote vascularistaion.
SDF, IGF1 are secreted.

Question 5

Ex or micro

Transfer of molecules

Answer 5

Transfer of molecules from MSC-derived via exosomoes or microvesciles

Question 6

Programmed cell death is good

MSC fitness and efferocytosis

Answer 6

Trigger phagocytotic mechanisms to clean out the area and go through efferocytosis mechanism which engulfs apotoic cells but has secondary regulators of immune modulation of a disorder or info happeing in a paticular disease state.

Question 7

ATMPs

Answer 7

Advanced therapy medicinal products, used for humans for treatment of disease and injury

Question 8

Cystic fibrosis examples

Gene therapy

Answer 8

Contains genes that lead to a therapeutic effect
Insert recombinat genes into the body to treat genetic disorders like CFTR gene delivered to the lung or intestine via a virus

Question 9

Somatic cell therapy medicines

Answer 9

Contains cells or tissues that have been manipulate to change their biological characteristics or cells or tissues not intended to be used for the same essential function in the body.

phenotype, dont have oth3r mechanims like MSC to become different cells

Question 10

Tissue engineered medicines

Answer 10

Contain cells or tissues modified to repair and regenerate or replace human tissue.

Question 11

Occular burns cause stem cell deficiency

Occular burns and the use of HOLOCLAR

Look at diagram in lecture

Answer 11

Human corneal epithelial cells containing stem cells for severe limbal stem cell deficiency
The limbals stem cells are grown in the lab and then transplanted onto the diseased cornea.

Question 12

Developing a cell therapy product from limbal stem cells diagram?

Answer 12

Limbal Biopsy: A small tissue sample is taken from a healthy eye’s limbus area.

Enzymatic Dissociation: The biopsy is treated with enzymes to separate the cells.

Primary Culturing: The dissociated cells are seeded onto a layer of irradiated mouse feeder cells, where they are allowed to multiply. Antibiotics are present during this stage to prevent contamination.

Expansion and Cryopreservation: The expanded cells from primary culture are frozen in cryovials, which allows for testing and additional manufacturing if necessary.

Thawing: When needed, the frozen cells are thawed for further processing.

Drug Substance Formation: A cell suspension of epithelial cells is created from the thawed cells, which are then seeded in a fibrin matrix. This is done without expression in paraclones.

Secondary Culturing: The drug substance is cultured a second time on a fibrin matrix along with irradiated mouse feeder cells, but without antibiotics at this stage.

Final Product:Discs of fibrin carrying the autologous cells are prepared and packaged.

Grafting:The final product is grafted into the injured eye to restore the limbal stem cells and thus repair the corneal epithelium.