INTEGRATED COURSE OF CELLULAR AND MICROBIAL BIOTECHNOLOGY - CELLULAR BIOTECHNOLOGY
- Overview
- Assessment methods
- Learning objectives
- Contents
- Delivery method
- Teaching methods
- Contacts/Info
The knowledge acquired during the course of Cytology and Histology and the basic techniques of Molecular Biology are fundamental.
Examination tests are scheduled as agreeded by the Degree Course. The test consists of an oral interview during which the report of the laboratory experience is discussed. This part is evaluated with a score between 0 and 6/30. During the interview, the student, in addition to demonstrate the knowledge / skills to work in a cell culture laboratory, must be able to plan an experiment involving the use of cell cultures and propose solutions to situations proposed by the professor. The oral exam will be evaluated with a score between 14 and 24/30. The final mark, expressed out of thirty, is determined by the sum of the two tests, and will take into account the relevance of the answers to the questions asked, the correct use of scientific terminology and the student's ability to present a general vision by establishing cross-connections between the various topics covered during the course.
The test is considered passed with a score of at least 18/30.
The course aims to provide the basic knowledge to work in a cell culture laboratory. Particular attention is paid to the preparation and maintenance of cell cultures whose application aspects will be studied in depth during the course. At the end, the student will have acquired theoretical and practical skills with reference to the use of cell cultures in the biological-biotechnological field.
The course is divided into two parts
1- LESSONS (40 h)
The topics covered concern:
1- Requirements of a cell culture laboratory in terms of equipment and precautions to be taken. History of cell cultures. Preparation of a cell culture, its maintenance, advantages and disadvantages in the various uses. Outline of optical and electronic microscopy.
2- Cell count by Burker and Neubauer, automatic cell counts. Separation of cell populations by clonal selection, differential centrifugation, antigen-antibody interaction, flow cytometer.
3- Outline of radiochemistry with particular attention to the principles of liquid and solid scintillation, evaluation of proliferation (BrdUTP and thymidine) and antigen-antibody interaction (RIA).
4- Immortalization by transfection and transformation techniques using plasmids, chemical and physical methods, or the use of viral vectors.
Control of cell contaminations (microbiological culture in broth and agar, PCR, DNA stain).
5- Control and evaluation of cell death, genotoxicity and gene expression by different methods: colorimetric, chemiluminescent, COMET, TUNEL, micronucleus tests and microarrays.
6- Set up of cytotoxicity tests (colorimetric, fluorescent, chemiluminescent)
7- Production, purification of monoclonal antibodies and examples of their application (ELISA).
8- Preparation, maintenance and use of primary stem cell cultures.
9- FETAX test and CAT assay on zebrafish.
10- Assisted reproduction techniques.
2- LABORATORY EXPERIENCE (12h)
During the laboratory experience the student will learn:
- to count a cell suspension and to evaluate the cell morphology
- to detect the presence of mycoplasma contamination by nested PCR
- to run a cytotoxicity and cell proliferation test using colorimetric assays
- to draw up the technical report
The teaching activity includes lectures and laboratory experiences.
• The lectures in the classroom will be supported with pptx presentations and the viewing of videos.
• The practical activity in the laboratory will be presented both during the frontal lessons and during the practical lessons. The experiences foresee the application of the guidelines to work safely with cell cultures. Each experience is described in the "laboratory notebook" which allows the student to easily carry out the proposed experiences. The organization requires each student to work independently.
The teacher is available to meet students for information on appointment by telephone or e-mail (rosalba.gornati@uninsubria.it).