Biotechnologies:experimental Models in Research

Unit 1: Cell culture
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
Educational visit in Italian laboratory

Unit 2: Alternative methods in Toxicology
Lectures
Toxicology, Alternative Methods and 3Rs Replace, Refine, Reduce (2 hours)
Alternative Methods and Validation (2 hours)
Replacement: validated in vitro test (4 hours)
Integrating Testing Strategy (1 hours)
Skin sensitization (2 hours)
Endocrine disruptor and alternative methods (1 hour)
Carcinogenesis (2 hours)
Reproductive Toxicology and Alternative methods (2 hours)
Botulin Toxin and 3Rs (1 hour)
Laboratory activity
In vitro markers in toxicity testing (2 hours)
In silico methods (4 hours)
ITS, Integrating Testing Strategy examples (2 hours)
Educational visit in Italian laboratory (4 hours)

Unit 3: Wellbeing and care of laboratory animals
Lectures:
Welfare and behaviour of laboratory animals
Husbandry and breeding of transgenic Rodents
Husbandry and breeding of Zebrafish (Danio rerio)
Husbandry and breeding of African clawed frog (Xenopus laevis)
Husbandry and breeding of Nematodes
Practical experiences:
Research facilities visit
Laboratory animals breeding facility visit