Program
Fundamentals of cell culture and biosafety
– Introduction: concepts of cell culture (objectives of in vitro cell culture, cell types, cell lines vs. primary cultures, media, supplements, and other reagents).
– Cell culture laboratory: equipment and workflow (laminar flow chambers, CO₂ incubators, phase contrast microscope; cleaning and maintenance).
– Biosafety and good practices: typical BSL2; PPE; good aseptic practices; waste classification and management.
– Basic cell culture techniques: subculture, counting (manual/automatic), seeding cells, cryopreservation, and thawing.
– Advanced models: spheroids vs. organoids; matrix requirements; differences from 2D.
– Contamination: types, prevention, detection, and response (bacterial/fungal, mycoplasma, cross-contamination; quality control routines).
– Good practices & troubleshooting.
Learning outcomes: acquire basic knowledge for performing cell culture procedures; identify risks and apply GLP/BSL; recognize signs of contamination and response plan.
Functional and molecular biology assays
– Viability, proliferation, and survival: MTT/XTT/Resazurin/ATP); doseresponse curves and IC50 calculation.
– Immunocytochemistry/immunofluorescence: marker labeling, negative controls/isotype, quantification concepts.
– Flow cytometry: principles and applications (labeling, basic protocols for assessing viability/apoptosis).
– Genotoxicity and cytotoxicity: comet assay, γH2AX, micronuclei.
– Migration and invasion: wound healing, Transwell®.
– Nucleic acid isolation: DNA/RNA from culture; quality/quantity.
Learning outcomes: choose and perform an assay according to scientific rationale; understand limitations and sources of bias; interpret results.
Experimental design and biostatistics
– Design of in vitro studies: controls, replicates (technical/biological), randomization, curves and normalization; reproducibility checklists.
– Essential biostatistics: types of data, value of n, confidence interval, parametric vs. nonparametric tests, results representation.
Learning outcomes: build a reproducible experimental plan; apply appropriate statistical analysis and report results.
Laboratory Practice
Objective: Apply the essential cell culture methodologies and techniques from modules 1-3 in a laboratory setting, ensuring good practices and reproducibility.
– Cell culture techniques: demonstration and practice
o Good practices
o Working under aseptic conditions
o Thawing and cryopreservation
o Subculture procedures
– Cell culture methods and applications: demonstration and practice
o Contamination detection
o Cell culture assays
o Troubleshooting
Assessment distribution
– Continuous practice (skills logbook) – 40%
– Mini-project: design of an in vitro study (2D or 3D), sampling plan, control, metrics, planned statistical analysis – 40%
– Theoretical quiz (15–20 questions) – 20%
Internship(s)
NAO