OVERVIEW

Studying a biological phenomenon involves choosing an appropriate study model, and in vitro cellular culture systems have proven to be very useful over the years for various experiments. In recent years, also with the aim of minimizing the use of animals in experimentation, various 3D culture systems have been developed that increasingly approximate the physiology of the organism. In particular, thanks to the development of technology, various types of three-dimensional structures are obtained on which cells can recreate an environment similar to an organ or tissue. Lastly, various innovative engineering systems have been developed to support cell culture, taking into account the different experimental needs.

In this course, various 3D cell culture systems applied to different areas of basic and applied biology study will be presented, providing the student with an updated bibliography.

AIMS AND CONTENT

LEARNING OUTCOMES

The aim of this course is to offer an overview of the main aspects of three-dimensional models of cell cultures, from spheroids to organoids and 3D-printing. Simulated microgravity models to support the formation of 3D structures and some examples of bioreactors and microfluidics, which allow the maintenance and control of cell culture for pharmacological and biomedicine studies, will also be presented.

AIMS AND LEARNING OUTCOMES

The aim of this course is to broaden knowledge on 3D culture systems, models that are increasingly used in both basic and applied research. In particular, the following will be covered:

• The "dynamic reciprocity" between the cell and the extracellular matrix in determining cell fate

• Modulation of microenvironment signals compared in 2D and 3D cultures.

• Definition of spheroid and organoid, differences, advantages, and disadvantages of the different types of 3D cultures. Their applications in various areas of biological research

• Biomaterials as a three-dimensional support for cell cultures

• Use of 3D-printing for the formation of tissue-specific organoids

• Use of organoids for the study of innervation and vascularization processes

• Use of bioreactors for dynamic cell cultures and basics of microfluidics and organ-on-chip

• Use of simulated microgravity models for the formation of 3D cellular structures: Random positioning machine (RPM), rotating wall vessel (RWV). Their application in ground-based research

• Review of scientific literature

• Laboratory activity for setting up a 3D cell culture

By the end of the course, the student will be able to:

• Understand the recent literature on various systems of three-dimensional cell cultures

• Understand the advantages and disadvantages of 3D cultures in various applications

• Have the basic knowledge to set up three-dimensional cell cultures in static and dynamic conditions, through the use of bioreactors and tools that aid the formation of three-dimensional cellular structures.

PREREQUISITES

Students will be required to have a basic understanding of chemistry, biochemistry, cell biology and 2D cell cultures.

TEACHING METHODS

24 hours of lectures and 16 hours of laboratory work.

Students with valid certifications for Specific Learning Disorders (SLD), disabilities, or other educational needs are invited to contact the professor and the disability coordinator of the School/Department at the beginning of the course to discuss any teaching methods that, while respecting the goals of the instruction, take into account individual learning methods.

SYLLABUS/CONTENT

Program of Lectures:

• The "dynamic reciprocity" between the cell and the extracellular matrix in determining cell fate
• Modulation of microenvironment signals such as cell adhesions, mechanical forces, and soluble factors, etc., compared in 2D and 3D cultures.
• 2D and 3D cultures: differences. Advantages and disadvantages
• Definition of spheroid and organoid, differences, advantages, and disadvantages of the different types of 3D cultures. Their applications in various areas of biological research
• Biomaterials as a three-dimensional support for cell cultures
• Use of 3D-printing for the formation of tissue-specific organoids
• Use of organoids for the study of innervation and vascularization processes
• Use of bioreactors for dynamic cell cultures and basics of microfluidics and organ-on-chip
• Use of simulated microgravity models for the formation of 3D cellular structures: Random positioning machine (RPM), rotating wall vessel (RWV). Their application in ground-based research
• Review of scientific literature

Laboratory Activities: 

• Observation of the morphological characteristics of different rigid scaffolds under the microscope for 3D cultures in static, dynamic, and perfusion conditions
• Preparation of a cell culture in a hydrogel and on rigid scaffolds, followed by cell staining for identification
• Preparation of a spheroid cell culture using the hanging drop technique
• Introduction to the printer and its components (laminar flow, UV rays, compressed air, touch screen)
• Practical demonstration of the preparation and use of printing tools (syringes, inks, cells, tips), setting printing parameters, printing a model, and crosslinking the ink with calcium chloride
• Observation of bioreactors for static, dynamic, and perfusion cultures present in the laboratory and used in various projects

RECOMMENDED READING/BIBLIOGRAPHY

Scientific literature.

TEACHERS AND EXAM BOARD

Exam Board

CHIARA GENTILI (President)

SARA TAVELLA (President)

LESSONS

Class schedule

The timetable for this course is available here: Portale EasyAcademy

EXAMS

EXAM DESCRIPTION

The examination takes place in a written format with questions covering all the topics discussed during the course. The exam consists of a multiple-choice test with 30 questions on the lectures and laboratory experience.

ASSESSMENT METHODS

The assessment will be based on the correctness of the answers provided during the final exam, which aims to verify the assimilation of the covered contents and the ability to link and apply the acquired knowledge.