Animal-free innovations

Question 1

For what can iPSCs be used?

Answer 1

• Cell therapies
• Toxicology tests
• Personalized medicine
• Drug development
• Disease modelling

Question 2

Name characteristics of hypertrophic cardiomyopathy.

Answer 2

• Impaired relaxation
• Metabolic shifts (energy metabolism, oxidative stress)
• Increased left-ventricular wall thickness
• Cardiomyocyte hypertrophy and disarrays
• Fibrosis
• Increased risk on heart failure and sudden cardiac death

Question 3

How can iPSCs be used to study the disease mechanism of hypertrophic cardiomyopathy?

Answer 3

iPSCs can be used to study:
- hypertrophy
- calcium influx
- electrophysiology of cardiomyocytes

Question 4

There are two challenges regarding the use of iPSCs. Name these.

Answer 4

• Biological variability
• Technical variablity

Question 5

What is meant with biological variability in iPSCs?

Answer 5

• Genetic background
• Age or sex of the donor
• Choice of control (isogenic controls)

Question 6

What is meant with technical variability in iPSCs?

Answer 6

• Reprogramming technique
• Differentiation protocol
• Purity of cells
• Culture conditions

Question 7

What are strategies to overcome variability or increase reliability?

Answer 7

• Standardizing methods for differentiation and readouts
• Genetic quality control (karyotyping)
• Function quality control (cell-specific markers, purity of cells, functional read-out dependending on the cell type)
• Include multiple lines of the same mutation
• Include multiple batches of differentiation

Question 8

What is a major drawback for differentiated iPSCs?

Answer 8

Immaturity in morphology, metabolism, function:
- reported in cell types connected to heart, kidneys, lungs, liver and brain

Question 9

What are consideration that need to be taken into account when designing disease modelling studies?

Answer 9

• Determine the goal of your assay (diagnostics, intervention, investigating disease mechanism)
• Mutation of interest
• How many lines?
• Which types of controls to choose?
• Which model mostly suits answering the research question (2D, 3D, coculture)?
• Which functional read-outs are of interest (link to clinical symptoms)?

Question 10

What is the problem with drug safety screening?

Answer 10

• Developing a new drug costs appr. 1.3 billion euros and takes 10-15 yrs.
• Cardiotoxicity is a large problem with newly developed drugs
• Animal models are 78% predictive for contractile changes
• Current cardiotoxicity drug screening is outdated

Question 11