502272 - LABORATORY OF CELLULAR METHODS

courses

ID:

502272

Duration (hours):

72

CFU:

SSD:

BIOLOGIA APPLICATA

Year:

2025

Date/time interval

Primo Semestre (29/09/2025 - 16/01/2026)

Course Objectives

Module 1: teacher Elena Maria Clotilde Raimondi - Learning of the techniques for somatic cell colture and for cytogenetic and molecular cytogenetic analysis. - Knowledge of the human karyotype and of the methods used for chromosome identification Module 2: teacher Sergio Comincini The module intends to provide the theoretical-practical knowledge of basic methodologies for cell maintenance and analysis together with the basic principles of optical microscopy, fluorescence and spectrophotometric analysis. Through these activities the student will acquire basic skills for the preparation of eukaryotic cell cultures, the analysis of their sub-cellular composition and functional microscopic evaluations of the alterations of the physiological state of eukaryotic cells. Module 3: Teacher Rossella Tricarico Learning Objectives -Provide the theoretical and practical foundations for designing functional assays on adherent cell cultures with short- and long-term endpoints. -Introduce principles of data analysis and critical interpretation. Learning Outcomes -Set up cell preparations on appropriate culture supports for specific functional assays. -Perform clonogenicity and senescence assays, as well as proliferation and migration assays, under basal conditions and following treatments or stress. -Acquire images and measurements, quantify relevant parameters, and present results in tabular/graphical form. -Interpret results, distinguishing between short- and long-term effects.

Course Prerequisites

Basics of Genetics, Cytology and Histology.

Teaching Methods

Laboratory exercises

For the categories of students who have the right to benefit from "inclusive teaching methods", the teachers will arrange ad personam meetings, in person and/or remotely, agreeing an exam program with the student.

Assessment Methods

Written test on the experiments carried out. Module 1: Two questions requiring a detailed answer and 10 questions with a short answer are asked. Module 2: 3 open-ended questions are asked. Module 3: 6 multiple-choice questions + 2 open-ended questions.

Texts

All the slides presented during the course are available for the students.

Modulo 1: teacher Elena Maria Clotilde Raimondi Mammalian cell cultures, tripsinization, counting and seeding of the cells,viability and proliferation assays, chromosome preparation,immunofluorescence, purification and labelling of DNA probes, in situ hybridization, experimental demonstratio of the semiconservative DNA replication (SCE assay), chromosome banding, human karyotype reconstruction, identification of the human chromosomes involved in the main chromosome pathologies, fluorescence microscopy analysis and use of CCD cameras. Module 2: teacher Sergio Comincini Isolation and analysis of extracellular vesicles. Modification of vesicular contents by introducing fluorescent molecules via electroporation techniques and subsequent administration of the modified vesicles to recipient cells. Observation of the cellular internalization process using a fluorescence microscope. Module 3: Teacher Rossella Tricarico Functional assessment of adherent cell cultures at short- and long-term endpoints, under physiological conditions and after treatments or stress: clonogenic assay (long-term survival), senescence assay (proliferation arrest), proliferation assay, and migration assay, with particular emphasis on data analysis and interpretation. Morphological observation across multiple cell lines (bright-field and fluorescence microscopy); preparation of cell specimens on different supports and in distinct experimental setups tailored to each assay. For each assay: image and measurement acquisition, quantification, and analysis of results.