Degree | Type | Year | Semester |
---|---|---|---|
2502445 Veterinary Medicine | OT | 5 | 0 |
CONTEXTUALIZATION:
Cell Culture in Biomedical Research is taught in the 2nd semester of the 5th year of the Veterinary degree in the Faculty of Veterinary Medicine. This is a subject with a certain degree of specialization. It is intended that the student acquires basic notions
regarding cell culture methodologies used for the manipulation and culture of mammalian eukaryotic cells. For this reason, the subject has a strong practical component.
This subject aims to provide the Veterinary student with the basic knowledge regarding the basic theoretical and practical knowledge to perform cell culture techniques of animal cells. The students will also learn techniques used for the characterization of
the cells and for the detection of cytotoxic and mutagenic effects of various compounds (medications, additives, etc.). These are techniques that are very useful in research labs and frequently used in derivative industries, such as pharmaceuticals.
OBJECTIVES:
1) To learn about the equipment and basic material of a cell culture laboratory
2) To learn the basic methodologies and protocols used to characterize mammal cells
THEORY PROGRAM
1. Introduction to cell culture: Basic review of cell biology: organelles and functions of the cell. History of cell culture. Applications of cell culture. Advantages / disadvantages of cell culture.
2. Equipment and aseptic environement: Cell culture room or laboratory, laboratory equipment, cell culture equipment, flow hoods, biological security cabinets, aseptic technique.
3. Establishment of a cell culture: Types of cell culture: monolayer, suspension, 3D. Tissue isolation, explant cell culture, primary cell culture, cell separation: centrifugation, cytometry, matrix, magnetism. Cellular requirements: environment,
media components, pH control, cell growth, cell count, viability assessment, cell proliferation, cryopreservation: freezing media, cryoprotectors and cooling rate. Establishment of a cell line.
4. Mortal and immortal cell lines: primary cell culture, cell transformation, cell immortalization, stem cells: types, culture, differentiation and applications. Hybridomas: immortal cells and antibodies synthesis.
5. Transfection: transfection, transduction and transformation. Viral transfection. Chemical and physical transfections. Advantages and disadvantages.
6. Cell characterization: cellular morphology, microscope types, fluorescence microscopy. DNA characterization: fluorescence in situ hybridization (DNA and RNA), DNA extraction, PCR, sequencing.
Protein characterization: fluorescence immunodetection of protein markers, western blot.
PRACTIC PROGRAMM
1. Maintenance and subculture of cell lines. Growth control of in vitr cultured cells. Morphology assessment.
2. Fixing and targeting of cells by immunofluorescence. Cellular characterization through antibodies.
3. Cytotoxicity tests: Control of cell viability and proliferation and toxicity testing: apoptosis and DNA damage.
4. Freezing and thawing of cell lines.
5. Discussion of results.
Title | Hours | ECTS | Learning Outcomes |
---|---|---|---|
Type: Directed | |||
Practical lessons | 16 | 0.64 | 3, 2 |
Theory lessons | 10 | 0.4 | 2 |
Type: Autonomous | |||
Individual study | 42.5 | 1.7 | 1, 2 |
Practical lessons report elaboration and presentation | 3 | 0.12 | 1, 3 |
Scientific paper presentation | 2 | 0.08 | 1, 3 |
Title | Weighting | Hours | ECTS | Learning Outcomes |
---|---|---|---|---|
Practical evaluation | 50% | 0.5 | 0.02 | 1, 3, 2 |
Theory evaluation | 50% | 1 | 0.04 | 1, 2 |
BIBLIOGRAPHY:
R.I. Freshney. Culture of Animal Cells: A manual of basic technique 6th Edition. John Wiley & Sons, 2010.
WEBS:
http://bcs.wiley.com/he-bcs/Books?action=index&bcsId=5959&itemId=0470528125
https://www.youtube.com/user/gibcocellculture/videos?view=0&sort=p&flow=grid