C8348 - IN VITRO MODELS AND ORGANOIDS

Learning outcomes

By the end of the course, the student understands the history and development of in vitro models, from HeLa cells to the generation of cellular spheroids and organoids. The student acquires knowledge of the main techniques for culturing cell lines, including the distinction between primary and immortalized cells, and understands and differentiates between 2D and 3D culture methods, as well as the current protocols for producing various types of spheroids and organoids. Additionally, the student is able to select, apply, and interpret appropriate experimental methods for in vitro cell culture, including both 2D and 3D approaches and the generation of multicellular organoids.

Course contents

This course is part of the Integrated Course “Regenerative Medicine”.

Upon completion of the Integrated Course unit, the student will be able to understand the history and development of in vitro models, from HeLa cells to the generation of cellular spheroids and organoids. The student acquires knowledge of the main techniques for culturing cell lines, including the distinction between primary and immortalized cells, and understands and differentiates between 2D and 3D culture methods, as well as the current protocols for producing various types of spheroids and organoids. Additionally, the student is able to select, apply, and interpret appropriate experimental methods for in vitro cell culture, including both 2D and 3D approaches and the generation of multicellular organoids.

For a better understanding of the contents it is recommended that the student has already acquired knowledge about: structure of the main biological macromolecules, knowledge of the biology and physiology of eukaryotic cells, calculations to carry out dilutions and to set up a PCR reaction, use the main instruments of the biological laboratory (micro-pipettes, centrifuges, optical microscope).

Module 1: Prof. Chiara Bernardini

The programme is as follows:

Theoretical lessons:

In vitro models: history and development of cell culture (2 hours)
Overview of the evolution of cell culture techniques and their contribution to biomedical and biotechnological research.

Role of cell cultures in animal biotechnology (1 hour)
Applications of cell culture systems in animal biotechnology, including research, diagnostics, and biotechnological production.

Cell types in culture: primary cultures and established cell lines (2 hours)
Characteristics, advantages, limitations, and applications of primary cell cultures and continuous (established) cell lines.

Cell culture methodologies: from two-dimensional (2D) to three-dimensional (3D) models (3 hours)
Principles, techniques, and applications of conventional 2D cultures and advanced 3D cell culture systems.

The cell culture laboratory: aseptic techniques and sterility (4 hours)
Laboratory organization, good cell culture practices, contamination prevention, aseptic techniques, and maintenance of sterile working conditions.

Management and maintenance of cell lines (4 hours)
Routine procedures for cell culture, including thawing, passaging, cryopreservation, quality control, and monitoring of cell health.

Module 2: Prof. Augusta Zannoni:

The programme is as follows:

Practical lessons:

Preparation of materials and reagents for animal cell culture (2 hours)
Preparation of culture media, reagents, and sterile laboratory materials required for the routine maintenance of animal cell lines, following good cell culture practices.

Thawing of an animal cell line (2 hours)
Procedures for the safe recovery of cryopreserved cell lines, including post-thaw handling and assessment of cell recovery.

Cell expansion, cell counting, and growth curve analysis (4 hours)
Routine subculturing procedures, determination of cell concentration and viability, and generation and interpretation of cell growth curves.

Assessment of cell viability and cryopreservation (4 hours)
Evaluation of cell viability using standard laboratory methods and application of protocols for the cryopreservation and long-term storage of animal cell lines.

Data analysis and interpretation of experimental results (4 hours)
Analysis, presentation, and critical interpretation of experimental data obtained during laboratory activities, with discussion of technical variability, sources of error, and the reliability of the results.

Readings/Bibliography

The teaching materials for this course are available on the Virtuale Learning Environment (https://virtuale.unibo.it/?lang=en ).

Teaching methods

The course includes both theoretical lectures and practical/laboratory sessions including lectures, group work, in-class exercises, lab work.

Considering the types of activities and teaching methods adopted, attendance for this course requires the successful completion of Modules 1 and 2 via e-learning, and Module 3 on health and safety training in study environments. Information about the schedule and access to Module 3 is available in the dedicated section of the Degree Program website.

Participation in practical and laboratory sessions requires wearing a lab coat and appropriate footwear. Suitable personal protective equipment (PPE), such as disposable latex gloves, will be provided as needed.

Assessment methods

Assessment methods

The final exam of the Integrated Course "Regenerative Medicine" consists of two parts.

The first part relative to the assessment for the Integrated Course unit “C8347 STEM CELLS” consists of a single test composed of:

• Multiple choice written test comprising 20 questions (13 regarding the contents of Module 1 and 7 the contents of Module 2). Each correct question will earn 1 point; no points will be subtracted in case of wrong answer or unanswered question.

• Open-ended written questions comprising 5 questions (3 regarding the contents of Module 1 and 2 the contents of Module 2). Each correct question will earn 2 points; no points will be subtracted in case of wrong answer or unanswered question.

The test will cover topics from all components of the Integrated Course, and is designed to assess the student's comprehensive knowledge of the subject matter. No supplementary materials or electronic devices (e.g., calculators, tablets, smartwatches, computers) may be used during the exam, except for those explicitly allowed by the instructor. If needed, a follow-up discussion will allow the Candidate to comment on the wrong answers. The exam is considered passed only if all parts are successfully completed. The final grade is determined by the sum of the results obtained in the two parts of the test.

The test assignment will last 1h and will be considered as successful with a minimum grade of 18/30.

The results of the written exam will be published within 5 working days on the Virtuale Learning Environment (https://virtuale.unibo.it/?lang=en ) by the designated course contact.

The second part relative to the assessment for the Integrated Course unit “C8348 - IN VITRO MODELS AND ORGANOIDS” consists of a single test composed of:

• Multiple choice written test comprising 15 questions (10 regarding the contents of Module 1 and 5 the contents of Module 2). Each correct question will earn 1 point; no points will be subtracted in case of wrong answer or unanswered question.

• Open-ended written questions comprising 3 questions (1 regarding the contents of Module 1 and 2 the contents of Module 2). Each correct question will earn 2 points; no points will be subtracted in case of wrong answer or unanswered question.

The test will cover topics from all components of the Integrated Course, and is designed to assess the student's comprehensive knowledge of the subject matter. No supplementary materials or electronic devices (e.g., calculators, tablets, smartwatches, computers) may be used during the exam, except for those explicitly allowed by the instructor. If needed, a follow-up discussion will allow the Candidate to comment on the wrong answers. The exam is considered passed only if all parts are successfully completed. The final grade is determined by the sum of the results obtained in the two parts of the test.

The test assignment will last 1h and will be considered as successful with a minimum grade of 18/30.

The results of the written exam will be published within 5 working days on the Virtuale Learning Environment (https://virtuale.unibo.it/?lang=en ) by the designated course contact.

The final exam of the Integrated Course "Regenerative Medicine" is considered passed only if two parts (Stem Cells and In vitro Models and organoids) are successfully completed. The final grade is determined by the average of the grades from the two parts of the exam, expressed out of 30. A minimum final grade of 18/30 is required.

With regard to the learning assessment, the use of AI is prohibited. Any use of AI constitutes a breach of academic integrity.

Teaching tools

Slides will be used together with videos; interactive software as Kahoot may be also used.

Teaching materials are available on the Virtuale platform and could be reached through the link on the teacher webpage.

In case of difficulty understanding the course content, the instructor is available for clarification meetings, which must be scheduled via email.

See the website of Chiara Bernardini

See the website of Augusta Zannoni

Office hours

See the website of Chiara Bernardini

See the website of Augusta Zannoni