INTEGRATED COURSE OF CELLULAR AND MICROBIAL BIOTECHNOLOGY - CELLULAR BIOTECHNOLOGY
- Assessment methods
- Learning objectives
- Delivery method
- Teaching methods
Thiss class requires the knowledge given during the lessons of Cytology and Histology toghetr with the rudiments of Molecular Biology tecniques.
Evaluation of the knowledges by an oral interview. The candidate has to demonstre to be able to work in a cell culture lab. In addition he/she has to set a cell culture experiment and propose alternative solutions to critical situations eventually proposed by the professor. During the interview, it will be also evaluated and discussed the quality of the report produced at the end of laboratory experience.
The final score will be the mathematical mean of the score acquired at the end of the interview (max 24/30) and that of the laboratory report (max 6/30).
The aim of the class is to provide the basic knowledge about the organization of a cell culture laboratory together with the comprehension of the most used applications in biotechnological field. This part which will be deepened during the laboratory practise.
The topics covered during the frontal lessons include:
1- Requirements for a cell culture laboratory in terms of instrumentation and precautions. History of cell cultures. Preparation of a cell culture, its maintenance, advantages and disadvantages. References concern optical and electronic microscopy (0.5 CFU - 4h).
2- Cell count using Burker and Neubauer, automatic cell counters and citofluorimeter. Separation of cellular populations by clonal selection, differential centrifugation, antigen-antibody interaction, cell sorting by citofluorimeter. (0.5 CFU - 4h).
3- Radioactive isotopes, liquid and solid scintillation, radioactive assays: evaluation of proliferation (radioctive thymidine) and antigen-antibody interaction (RIA and IRMA).
Transfection and transformation techniques with plasmids and other chemical and physical methods. Use of viral and non-viral vectors. (1 CFU -8h).
4- Immortalization control of contaminations (microbiological culture in broth and agar, PCR, DNA stain) (0.25 CFU - 2h).
5- Control and evaluation of cell death, genotoxicity and gene expression: colorimetric and chemiluminescent methods, COMET assay, micronucleus test, microarray (0.25 CFU - 2h).
6- Cytotoxicity tests and assessment methods including the use of radioisotopes (1 CFU - 8h).
7- Production, purification and quantification of monoclonal antibodies. Non radioactive assays (ELISA). (0.25 CFU - 2h).
8- Preparation and maintenance of primary stem cell cultures (0.25 CFU - 2h).
9- FETAX Test and ETF Assay on zebrafish
10- Male and female reproductive apparatus, differentiation of male and female gametes (0.5 CFU - 4h).
11- Assisted fertilization techniques (0.25 CFU - 2h).
12- Preparatory lesson for laboratory practise: how to set up a cytotoxicity test from the choice of cell culture to the methods of evaluation and data analysis (0.25 CFU - 2h).
Program of laboratory practice:
Control of Mycoplasma contamination by PCR, cell counting by Burker chamber, preparation of a cytotoxicity test ans evaluation by MTT assay, evaluation of proliferation and cellular morphology by colorimetric assays and microscopic observation.
The recommended books are:
Freshney R.I. “Culture of animal cells. A manual of basic technique” Wiley-Liss, Inc.
Brown T.A. “Biotecnologie molecolari. Principi e tecniche” Zanichelli.
Alberts B., Johnson A., Lewis J., et al. “Biologia molecolare della cellula” Zanichelli.
The teaching support is periodically updated and consists of the slides that are shown by the professor during the class. These supports are available to students on the e-learning platform.
The course includes lectures (40 hrs) and laboratory training (16 hrs).
For what concern lectures, topic will be illustrated with the help of slides.
The laboratory training will be held by the teacher and two assistants.Students have to attendance all the laboratory course and eventual absences have to be justified.