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1. COURSE DECRIPTION – GENERAL INFORMATION Nataša Bauer, Maja Matulić 1.1. Course teacher 1.2. Name of the course Animal and plant cell culture 1.3. Associate teachers 1.4. Study programme (undergraduate, graduate, integrated) undergraduate obligate 1.5. Status of the course 2. COURSE DESCRIPTION 2.1. Course objectives 2.2. Enrolment requirements and required entry competences for the course 2.3. Learning outcomes at the level of the study programme to which the course contributes 2.4. Expected learning outcomes at the level of the course (4-10 learning outcomes) 1.6. Year and semester of study 1.7. Credit value (ECTS) 1.8. Type of instruction (number of hours L+S+E+e-learning) 1.9. Expected enrolment in the course 3rd y of undergraduate study of molecular biology 5 1+0+2 30-40 1.10. Level of use of e-learning (1, 2, 3 level), percentage of instruction in the course on line (20% maximum) Introduction to basic techniques in plant and animal cell culture cell biology theoretical and practical knowledge of the basic techniques in plant and animal cell culture Understanding principals of growth and development of cells and tissues Understanding the possibilities and obstacles during cell growth and development manipulation Ability of independent work on establishment and directioning of cell culture growth Ability of genetic manipulation with cells and tissues Animal cell culture Introduction in animal cell culture: what is animal cell culture and where is it used today, comparison of cell culture with model organisms, history of cell culture Conditions of cell culturing: types of cell culture medium and its components, , types of substrates for cell growth, contamination, buffers used for pH conditioning, cryopreservation t types of cell lines, growth curves 2.5. Course content broken down in detail by weekly class schedule (syllabus) Growth of cell colonies, methods of colony isolation, survival curves Primary cell culture: preparation of primary culture, methods of separation of different cell types, “evolution” of the cell line Methods of cell quantitation: cell counting, determination of the cell viability by differential staining, flow cytometry . Cell authentication and characterization: methods used for differentiating cells from diverse organisms, tissues and entities 1 Cell growth synchronisation: mitotic selection, synchronisation in G1 phase, inhibitors of S phase, double block in synchronization, methods for evaluation of cell synchrony New techniques in cell culture. Cell fusion and differentiation. Basic characteristics of cancer cell growth in culture and senescence Plant cell and tissue culture: Introduction to plant cell culture: what is plant cell and tissue culture and where is it used today. Structural, physiological and biochemical aspects of organogenesis in vitro. Characterization of callus tissue culture and patterns of growth in tissue cultures. Establishment and use of suspension cultures. Establishment and manipulation of protoplasts. Organ culture. Micropropagation and regeneration pathways and principles of vegetative propagation in vitro. Methods used in transformation of plant cells. Regeneration of transgenic tissues and plants. Somaclonal variation in cell cultures. Long term culturing of plant cells and cryopreservation. Exercises Animal cells: Introduction to cell culture laboratory and work in cell culture. Trypsinizing and counting cells. Seeding cells for growth curve and analysis of cell growth in culture by counting. Calculating the duplication time Primary cell culture: preparation of fibroblast culture from mouse skin. Explantation technique and preparation of cell suspension by trypsinization Influence of chemotherapeutics on the cell growth: growth curve obtained by counting the cells Influence of chemotherapeutics on the cell growth: survival curve obtained by counting colonies Cell synchronization: mitotic selection. Plant culture: Preparation of culture media. Sterilization of plant tissues, media and supplies. Sterile manipulation of cells and tissues. Selection and determination of transgenic plants. 2 Micropropagation by nodal segments. Micropropagation by adventitious buds. Suspension culture: determination of cell growth and viability. Final analysis and discussion of results. 2.6. Type of instruction x lectures seminars and workshops x exercises online in entirety mixed e-learning field work independent study multimedia and the internet x laboratory work with the mentor 2.7. Comments: (other) 2.8. Student responsibilities 2.9. Screening of student’s work (specify the proportion of ECTS credits for each activity so that the total number of CTS credits is equal to the credit value of the course)): 2.1. Grading and evaluation of student work over the course of instruction and at a final exam Class attendance Research Experimental work Report 0.2 Essay Seminar essay Tests 1.9 Oral exam 1 Written exam 1.9 Project partial written exams, class attendance, reports on laboratory exercizes Title Allberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P (2002): Molecular biology of the cell, Garland Publishing, New York. 2.2. Required literature (available at the library and via other media) 2.12. Optional literature (at the time of the submission of the study programme proposal) 2.13. Methods of monitoring quality that ensure acquisition of exit Animal cell culture exercizes, script scientific papers from relevant journals Practical training (Other--describe) (Other—describe) (Other—describe) Number of copies at the library 3 Availability via other media x x x Masters J. R. W. (2000): Animal cell culture 3. iz. Oxford University Press, Oxford Butler M (2004): Animal cell culture & technology, 2. iz. Bios Scientific Publishers, London i New York Freshney R. I. (2000): Culture of animal cells: A manual of basic technique. 4. iz. Willey-Lys, J. Willey & Sons. New York partial and final written exam, oral exam, laboratory exercizes and final report 3 competences 4