Fundamentals of Cellular and Tissue Engineering

Course guarantor:

Lecturer:

Tutor:

Supervisor:

Department of Biomedical Technology

Synopsis:

The aim of the subject is to acquaint students with the basics and methods of working with cell culture and their use both as a model replacement for in-vivo experiments and also as a means of preparing artificial tissue and organ replacements using tissue engineering methods. From the point of view of basic procedures, the issue of obtaining cell cultures of various phenotypes, suitable cultivation conditions, cultivation media, and instrumentation will be addressed. Substrates, biomaterials and decellularized carriers suitable for cell growth, the use of culture systems and bioreactors in terms of simulating physiological conditions and their contribution to supporting cell proliferation and differentiation for the needs of developing artificial tissue and organ replacements and 3D bioprinting technology will be addressed for application. The subject will also address the issue of legislation and regulatory conditions in connection with the use of tissue engineering for the preparation of artificial substitutes and modern therapy preparations.

As part of practical laboratory exercises, topics related to the basics of working with cell culture, working in sterile conditions will be implemented; preparation of culture media; imaging of cell morphology using fluorescence microscopy and histological staining; cell culture isolation; preparation of two- and three-dimensional carriers based on nanofibers and hydrogels and their deployment in cultivation bioreactors; 3D bioprinting.

Requirements:

Form of verification of study results: Credit - attendance at exercises and keeping a laboratory diary. Written exam in the form of a test (10 questions with multiple correct options) and 4 methodical examples. Students can have their own lab journal and lecture notes for the exam.

Student requirements: Mandatory attendance at laboratory exercises, keeping a laboratory diary for the given laboratory experiments. The laboratory journal replaces the protocol and is a condition for obtaining credit.

Syllabus of lectures:

1. Cell cultures, meaning, history, possibility of use in regenerative medicine, necessary equipment.

2. Obtaining cell culture from tissue, primary culture, cell line, stem cell and their characterization.

3. Substrates for cell growth, culture media, growth factors and their influence on culture development.

4. Tissue engineering, biomaterials, artificial substitutes with cell culture, decellularization, 3D bioprinting.

5. Cultivation systems and bioreactors, dynamic cell cultivation, stimulation of cell culture to support proliferation and differentiation.

6. Legislation associated with tissue engineering, modern therapy preparations, GMP/GLP.

Syllabus of tutorials:

1. Preparation of space for working in sterile conditions, preparation and sterilization of media and other solutions, basics of working in a laminar box.

2. Passage of cell culture, deployment of adherent culture on a planar carrier, stocking density.

3. Sample fixation, membrane permeabilization, antibody blocking, fluorescent labeling of nuclei.

4. Immunofluorescence staining using primary and secondary antibodies, imaging by fluorescence microscopy.

5. Preparation of a composite 3D carrier based on nanofibers and collagen.

6. Preparation of hydrogel with incorporated cell culture and deployment as a 3D substrate.

7. Isolation of cell culture from tissue – stem cells from adipose tissue.

8. Model preparation and subsequent 3D bioprinting of cell culture on a prepared nanofibrous carrier.

9. Preparation and installation of the substrate in the pressure stimulation reactor.

10. Preparation and placement of the substrate in the flow reactor, cultivation in shear stress.

11. Tissue decellularization.

12. Histological staining of the specimen, deparaffinization, HE staining.

Study Objective:

Study materials:

BRONZINO, Joseph D. The Biomedical Engineering Handbook. First edition, Boca Raton: CRC Press, 2000. ISBN 0-8493-0461-0.

VEMURI, Mohan C., CHASEm Lucas G. a Mahendra S. RAO. Mesenchymal stem cell assays and applications. First edition, Humana Press/Springer, 2011. ISBN 978-1-60761-998-7.

Note:

Further information:

No time-table has been prepared for this course

The course is a part of the following study plans:

• Biomedical Laboratory Methods (compulsory course)