Biotechnologies: experimental models in research

A.Y. 2019/2020
12
Max ECTS
102
Overall hours
SSD
AGR/18 AGR/20 VET/07
Language
English
Learning objectives
The aim of the course is to present basic and advanced methodologies in the field of biotechnological research. Several class activities will give students useful tools to develop a critical ability in identifying and choosing experimental models in relation to their applicability.
Expected learning outcomes
At the end of the course, the student will know:  the main basic and advanced methodologies in the research field and will acquired the skills allowing him to identify the most suitable experimental models for biotechnological research. Specifically, students will acquire knowledge on:
cell culture techniques
3Rs in toxicology and Replacement
alternative methods in toxicology and application
lab animals wellbeing, management and rearing techniques
Course syllabus and organization

Single session

Responsible
Lesson period
Second semester
Prerequisites for admission
Basic knowledge in toxicology is required.
Assessment methods and Criteria
The exam consists of a written evaluation aimed to evaluate knowledge and understanding of the lectures' subjects and of the laboratory activities. The test will be evaluated in thirtieths (minimum grade is 18 and the maximum is 30 "cum laude", with honors). The final score of course is the sum of the scores of the three teaching units. The score will take into account the accuracy and the capacity to motivate the answers, as well as the capacity to explain the concepts acquired with appropriate and updated terminology.
Mod. 1: Colture cellulari
Course syllabus
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
Teaching methods
Lectures and laboratory activity
Teaching Resources
Didactic material available on ARIEL: https://fchelicc.ariel.ctu.unimi.it/v5/home/Default.aspx
Culture of Animal Cells: A Manual of Basic Technique. R. Ian Freshney, Wiley-Liss, 2000.
Animal Cell Culture: A Practical Approach. John R. W. Masters, Oxford University Press, 2000.
Animal Cell Culture and Technology. Mike Butler, Michael Butler, Taylor & Francis, Inc., 2004.
Introduzione alle colture cellulari, Mariotti GL et al., Tecniche Nuove, 2010
Mod. 2: Metodi alternativi in tossicologia
Course syllabus
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)
Teaching methods
Lectures and laboratory activity
Teaching Resources
Mod. 3: Benessere dell'animale da laboratorio "Animal care"
Course syllabus
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)
Teaching methods
Lectures and tutorial activity
Teaching Resources
Didactic material available at www.fabioluzi.it
Mod. 1: Colture cellulari
AGR/18 - ANIMAL NUTRITION AND FEEDING - University credits: 5
Practicals: 36 hours
Lessons: 12 hours
Mod. 2: Metodi alternativi in tossicologia
VET/07 - VETERINARY PHARMACOLOGY AND TOXICOLOGY - University credits: 4
Practicals: 12 hours
Lessons: 18 hours
Professor: Caloni Francesca
Mod. 3: Benessere dell'animale da laboratorio "Animal care"
AGR/20 - AQUACULTURE, POULTRY AND RABBIT SCIENCE - University credits: 3
Practicals: 12 hours
Lessons: 12 hours
Professor: Luzi Fabio