LEARNING OUTCOMES

Provide students with an in-depth study of the techniques used for the growth and manipulation of cells "in vitro" and the use of different types of cell culture.

AIMS AND LEARNING OUTCOMES

Participation in the planned training activities (lectures and laboratory) will allow the students of the Biological Sciences Course to acquire the general knowledge and the fundamental notions for cell culture, which are necessary for the professional outlets in the Biological Sciences field.

The course aims to describe the basic principles of cell cultures and their use in many fields of experimentation and diagnostics. The course includes a theoretical part that illustrates the use of the types of cells that can be grown in a laboratory, the composition of the culture media, the methods of sterilization and the architecture of a cell culture laboratory, and a practical part where the individual participants will experience working in sterile conditions and develop basic cell counting protocols, determination of cell viability and wound healing.

The student must be able to:

• Knowing the principles that dictate the correct practice of cell cultures;

• Acquiring practical skills in working in sterile conditions, cell manipulation and measurement of some parameters;

• Understanding the purposes of cell culture practice.

Skills acquired at the end of the course: At the end of the course, the student must  acquire knowledge of the basic requirements that allow him to grow cells successfully and safely.

PREREQUISITES

Knowledge of the Biochemistry and Physiology programs.

Knowledge of the main aspects of General Biology with particular regard to the structure of the cell and its components.

TEACHING METHODS

The teaching consists of lectures and a part of a theoretical-practical laboratory. Attendance at classes and lab exercises is strongly recommended.

The laboratory exercises are aimed at the deepening of some basic methods for the practice of cell cultures.

In particular, small group exercises will be developed (3 students max) for the determination of:

 - Trypsinization;

- Cell count with trypan blue; 

 - Cell seeding;

 - Familiarization with the calculation for dilutions;

- Cell viability following different treatments;

 - Wound healing following different treatments;

The exercises are carried out in the DISTAV equipped laboratories (with optical microscopes, spectrophotometers, spectrofluorimeters, etc.).

SYLLABUS/CONTENT

In vitro cells. Primary cultures, finite and continuous cell lines. Basic techniques of cell cultures in vitro. Advantages and disadvantages of cell cultures. The 3 Rs. Necessary equipment and sterilization methods.

Composition of different media. Trypsinization and propagation of cell culture. Freezing and thawing. Cell adhesion. Kinetics of growth. Cellular count. Evaluation methods of cell proliferation, survival and toxicity. Viability test. Some applications: wound healing.

Architecture of a laboratory and safety regulations.

The Laboratory credit will be dedicated to the practical application of what was explained during the theoretical lessons (setting up a culture, cell count, subculture, wound healing).

RECOMMENDED READING/BIBLIOGRAPHY

• G.L. Mariottini et al. Introduzione alle colture cellulari: metodiche relative – MORGAN ed. tecniche
• Biochimica e biologia molecolare. Principi e tecniche
• Keith Wilson, John Walker Ed. RaffaelloCortina
• Provided by the teacher