Biotechnologies: Experimental Models in Research

Progress in cell culture technology allowed the development of "high throughput cell-based assays" widely used in many different disciplines. The module objectives are to provide an introduction to basic principles of cell culture technology and to present some specific advanced applications of cell culture technology to the field of animal nutrition, feed safety and quality and feed bioactive compound activity evaluation.

Lectures
Design and equipment for a cell culture laboratory
Cell Culture - basic principles
- Main types of cell culture: primary cultures and cell lines
- The cell environment: sterile environment, pH, temperature and supply of nutrients for growth
- Basic techniques for cell culture: suspension culture, monolayer culture, 3D culture
- Advanced technique for cell culture: insert, scaffold and mammosphere
- Quality control
- Protocols for cell quantification and viability
Cell culture protocols and application to the field of animal nutrition
- Cell culture protocols for research: differentiation, tissue remodelling and nutrigenomic
- Epithelial cell barrier: trans epithelia transfer of nutrients, feed bioactive compound activity
- Cell culture for evaluation of feed functional and health claims

Journal club
Critically evaluation of recent articles in scientific literature (work group)

Exercises:
- The "Do's and Don'ts" of cell culture: aseptic technique and good cell culture practice
- Preparation and storage of culture media
- Cryopreservation and storage of cell culture
- Cytotoxicity and viability assay
- Culture of cell lines
- Cell-based assays for in vitro testing of functional properties of feed bioactive compounds and additives: probiotic adhesion tests
- National cell culture laboratories and cell bank: visit

Lectures
Toxicology, Alternative Methods and 3Rs Replace, Refine, Reduce (2 hours)
Alternative Methods and Validation (2 hours)
Replacement: validated in vitro test (4 ore)
Integrating Testing Strathegy (2 ore)
Skin sensitization LLNA (2ore)
Carcinogenesis (2 hours)
Reproductive Toxicology and Alternative methods (2 ore)
Botulin Toxin and 3Rs (2 ore)

Practical
In vitro markers in toxicity testing (4 hours)
In silico methods (4 hours)
ITS, Integrating Testing Strathegy examples (2 hours)
Bioreactors (2 hours)

Lectures

Management and breeding of transgenic colonies of Rodents (2 hours)
Management and breeding of Zebrafih (Danio rerio) (2 hours)
Anesthesia, analgesia and euthanasia: D.L. 4 marzo 2014 n. 26 (2 hours)
Dosing and sampling (2 hours)
The experimental protocol in biomedical research (2 hours)
Adoptions (1 hour)
Nematodes in research (1 hour)

Tutorials

Non-invasive imaging techniques: Thermography (2 hours)
Murine models (3 hours)
Visit a research facility (4 hours)
Visits to breeding of laboratory animals (3 hours)