Material provided by the teacher and present on the e-learning platform
of the university (access of students enrolled in the course through the
own credentials)
Learning Objectives
The aim is to transmit updated information on the methods of culture and handling of yeasts, insect cells, mammalian cell lines, stem cells and tumor cells, in order to learn their main biotechnological fields of application
Prerequisites
Good knowledge of the topics of general biology and molecular biology is required
Teaching Methods
Lectures and lab exercises
Type of Assessment
Oral examination
Course program
Introduction to different cell systems: yeasts, insect cells, human cell lines, stem cells, tumor cells. Maintenance of cultures in vitro, in suspension and in adhesion. Stem cells: maintenance, supports, co-cultures, growth factors, molecular mechanisms of differentiation, embryonic stem cells (ESC), somatic (ASC), induced pluripotent stem cells (iPSC). Cultures on three-dimensional supports, spheroids, organoids and their applications. Yeast: growth and maintenance conditions, genetic manipulation, expression of recombinant proteins, induction systems. Insect cells: characteristics and growth, genetic manipulation through baculovirus infection, protein expression. Mammalian cell lines: growth and maintenance, transfection techniques. Expression methods in mammalian cells: constitutive and inducible promoters, Tet-On / Tet-Off, expression of recombinant proteins, secretion, post-translational modifications. Methods for selecting stable cell lines, selection markers, fluorescent reporter. Genetic manipulation by use of transposons. Genome editing techniques: zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), CRISPR / Cas9 system.