Cytotoxicity evaluation of chemical compounds

Basic knowledge in biochemistry, in vitro cell culture and cytophysiology.

Contact hours (32 h):

- lectures - 10 h

- laboratory classes – 20 h

- consultations with teacher - 2 h

Student's individual work (68 h):

- preparation to laboratory classes – 8 h

- writing projects and reports – 10 h

- repetition and extended study - 20 h

- preparation to pass the lab test - 10 h

- preparation to pass the final test – 20 h

Total time 100 h = 4 ECTS

W1: Student describes the important concepts and processes in biochemistry, cell biology and toxicology K_W01, K_W08, K_W15

W2: Student demonstrates the knowledge on selected cytotoxic agents and characterizes methods of their identification and evaluation such as viability or survival tests, assays based on cell proliferation, metabolic activity, microtitration assays and drug interactions K_W02, K_W11

W3: Student uses the terminology specific for toxicology and defines: viability, toxicity, survival, tissue and systemic responses, genotoxicity, and inflammation response K_W02, K_W06

W4: Student defines the research hypothesis and can choose adequate methods/procedures to solve the scientific problem related to detection of potential cytotoxic effect(s) K_W11, K_W12, K_W17, K_W21

W5: Student has the knowledge on preparation of presentations, reports, documentation and evaluation of obtained results, can draw correct conclusions K_W20

U1: Student has the ability to work individually or in a group K_U02

U2: Student uses the knowledge on structure and properties of biomolecules and molecular markers to diagnostic analyses K_U04, K_U05, K_U10

U3: Student uses specialist terminology and biological nomenclature and specialized terms in biochemistry, biotechnology and toxicology K_U13

U4: Student recognizes the possibilities and limitations of molecular diagnostics techniques in cytotoxicity assays K_U10, K_U11, K_U12

U5: Student applies the correct and suitable cytotoxicity assay to determine potential cytotoxicity of compounds being studied K_U05, K_U12

U6: Based on the analyzes and experiments performed, student identifies the effects of cytotoxic influence at the cellular, tissue, organ and whole organism level K_U17, K_U19

U7: Student interprets correctly obtained results, draws the final conclusions and can prepare reports, laboratory documentation and expert presentations using correct scientific and technical terminology K_U13, K_U14

K1: Student understands that the lifelong learning is the ongoing, voluntary and self-motivated pursuit of knowledge for either personal or professional reasons K_K05

K2: Student is aware of the responsibility of his work as diagnostic professional and takes care of developing the positive image of this profession K_K08

K3: Student can work in a team and/or coordinate the work of research group K_K09

K4: Student can critically evaluate the obtained results K_K10

Lecture: informative lecture

Laboratory classes: students (2-3 persons in a team) have to perform the analyses according to the obtained protocol. The experiments have to be conducted in aseptic conditions therefore lab groups should be as small as possible (max. 8-10 persons).

- wykład informacyjny (konwencjonalny)
- wykład problemowy

- ćwiczeniowa
- doświadczeń
- laboratoryjna
- obserwacji
- projektu

The lecture presents selected issues from the field of cytotoxic influence of different compounds on the cytophysiological processes as well as methods and regulations in the cytotoxicity evaluation.

Laboratory classes will make students familiar with the methods of cells selection and preparation for cytotoxicity analyses and with commonly used cytotoxicity tests. Based on the analyzes and experiments performed, students will identify the effects of cytotoxic influence.

Lecture:

1. Basics of in vitro cell culture.

2. Morphological and metabolic changes after exposure to cytotoxic agents.

3. Disorders in cell cycle, genotoxicity and inflammatory responses.

4. Cellular senescence and death.

5. Viability, toxicity and survival – nature of cytotoxicity assays.

6. The national and international standards and regulations in cytotoxicity evaluation (e.g. ISO 10993).

Laboratory classes:

1. Cell isolation and culture for cytotoxic analyses.

2. Tests assessing cell metabolism and membranes function.

3. Integrity of cell membrane as an important parameter in cytotoxicity assays.

4. Oxidative stress damage. Determination of reactive oxygen species.

5. Cellular senescence assays.

6. DNA damage and genotoxicity.

7. Experiment summary and final test.

There is not a single academic book, which covers the whole subject content. Lectures and instructions for laboratory classes will be the basis for the preparation to the final tests. Some of the issues can be found in the following books:

1. Stokłosowa S. „Hodowla komórek i tkanek” (PWN, Warszawa 2011)

2. Freshney R. I. „Culture of Animal Cells: A Manual of Basic Technique” (Wiley, Nowy Jork 2005)

3. Kilarski W. „Strukturalne podstawy biologii komórki” (Wydawnictwo Naukowe PWN, Warszawa 2003)

4. Kawiak J. „Seminaria z cytofizjologii dla studentów medycyny, weterynarii i biologii” (Elsevier Urban&Partner, Wrocław 2002)

Lecture:

Test written exam checking effects K_W01, K_W02, K_W06, K_W08, K_W11, K_W12, K_W15, K_W17, K_W20, K_W21. Test consisting of single-choice questions offering 5 eventualities and open questions. Each correct answer - 2 points. At least 20 questions in the test. Passing the exam after reaching at least 50% of the points available. Very good mark for more than 90% of the points. Other grades proportionally in the 50-90% range.

Laboratory classes:

Student activity and preparation to classes (K_U04, K_U05, K_U10, K_K5) – 20% of the final score

Oral presentation on chosen topic (K_U13, K_U14) – 30%

Final test (K_U04, K_U05, K_U10, K_U13) consisting of single-choice questions offering 5 eventualities - 50% of the final score. Each correct answer - 2 points. At least 10 questions in the test. Passing the test after reaching at least 50% of the points available. Very good mark for more than 90% of the points. Other grades proportionally in the 50-90% range.

not applicable