Biotechnologies:experimental models in research

A.Y. 2020/2021
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
Didactic methods
Lessons will be held on the Microsoft Teams/Adobe Connect platform and can be followed synchronously on the basis of the second semester timetable, if still necessary. Lesson PDF files and teaching material may be available on the Ariel platform.

Program and reference material
The program and reference material will not be changed as far as possible

Learning assessment procedures and assessment criteria
The exam will take place orally using the Microsoft Teams platform.
Course syllabus
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
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)
TutorialsNon-invasive imaging techniques: Thermography (2 hours)
Murine models (3 hours)
Visit a research facility (4 hours)
Visits to breeding of laboratory animals (3 hours)
Prerequisites for admission
Basic knowledge in toxicology is required.
Teaching methods
Lectures, laboratory and tutorial activity
Teaching Resources
Unit 1
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

Unit 2
Didactic material available on ARIEL https://fcalonimat.ariel.ctu.unimi.it/v5/home/Default.aspx

Unit 3
Didactic material available at www.fabioluzi.it
Assessment methods and Criteria
The exam is aimed at:
- ascertain the achievement of the objectives
- ascertain the ability to apply knowledge and understanding
- verify the student's independence of judgment
- ascertain the command of the specific language
Unit: Alternative methods in Toxicology
VET/07 - VETERINARY PHARMACOLOGY AND TOXICOLOGY - University credits: 4
Practicals: 12 hours
Lessons: 18 hours
Professor: Caloni Francesca
Unit: Cell culture
AGR/18 - ANIMAL NUTRITION AND FEEDING - University credits: 5
Practicals: 36 hours
Lessons: 12 hours
Professor: Cheli Federica
Unit: Wellbeing and care of laboratory animals
AGR/20 - AQUACULTURE, POULTRY AND RABBIT SCIENCE - University credits: 3
Practicals: 12 hours
Lessons: 12 hours
Professor: Luzi Fabio