PREREQUISITI E PROPEDEUTICITÀ

Conoscenze di base di biologia cellulare e  molecolare,

Organ physiopathology,

Epigenetics and stem cells.

CONOSCENZE E ABILITÀ DA ACQUISIRE

Objective of this series of lessons is to introduce students to the concepts of stem cells, tissue healing and regeneration. The first module will be specifically focused on the cardiovascular system and will discuss: the involvement of stem cells in cardiovascular development and physiology, the experimental use of stem cells for cardiovascular healing and regeneration, and clinical trials experimenting the use of stem cells for the treatment of acute and chronic heart disease.

A particular emphasis will be given to review literature data, pointing out to seemingly divergent results. At the end of this module, students should be able to critically revise regenerative medicine studies trying to reconcile seemingly contrasting results or disproving them.

Students that will develop a special interest in these branch of biotechnology, will be given the opportunity to perform supervised training in our research laboratories.

The second module will illustrate how microscopy-, flow-cytometry-, cell separation-, cell culture-, and nanotechnological-techniques can impact both in research and diagnostic practice. At the end of this module, students must be able to chose among the different in vitro and in vivo assays the most proper for a specific experimental or diagnostic task. Additionally, they are required to understand the advantages and limits of each assay.

Last, students will be introduced to a very relevant clinical issue related to organ transplantation, that is organ preservation, especially in the scenario of older and ‘higher risk’ donors. Specifically, at the end of this series of lectures, students must know the main mechanisms of organ injury associated with organ preservation and advanced methodologies that have been experimented to improve it.

PROGRAMMA/CONTENUTI DELL’INSEGNAMENTO

MODULO:

1. Stem Cells and Regenerative Medicine
1. Define tissue healing, distinguishing regeneration from repair and fibrosis,
2. discuss examples of tissue regeneration, providing evidence for the possible involvement of stem cells and the immune system in these processes,
3. define the fundamental in vivo and in vitro properties of stem cells,
4. present the major models that have been put forward to characterize stem cell proliferation and differentiation properties,
5. discuss fundamental stem cell properties, such as asymmetric cell division, self-renewal, potency, quiescence, and dysfunction,
6. introduce the concept of stem cell niches and the effects exerted by the microenvironment (both soluble factors and physical forces) in regulating stem cell fate,
7. discuss the identification, properties, and growth requirements of Embryonic Carcinoma (EC) and Embryonic Stem (ES) cells,
8. present the molecular basis for pluripotency and explain the discovery of induced Pluripotent Stem cells (iPS),
9. briefly introduce heart disease, underscoring the importance of atherosclerosis and coronaropathy in the pathogenesis of ischemic cardiomyopathies,
10. introduce the importance of angiogenesis in the prevention of ischemic heart disease,
11. present evidence on the existence of a postnatal cardiomyocyte renewal, the involvement of stem cells in cardiogenesis, and in postnatal cardiomyocyte turnover,
12. document the experimental use of cardiac resident stem cells for myocardial repair,
13. discuss how to drive the differentiation of pluripotent cells towards cardiomyocyte lineages, briefly delineating the transcription factor network governing cardiogenesis,
14. discuss the main features of the three major stem cell classes present in the bone marrow; hematopoietic stem cells, marrow stromal cells and endothelial progenitor cells,
15. present data on the experimental use of bone marrow derived cells for myocardial repair,
16. summarize data obtained in clinical trials experimenting the use of different stem cell classes for myocardial repair, offering a critical perspective on the main findings,
17. discuss critical issues that need to be addressed to ameliorate cell based therapies, such as delivery route, engraftment, integration of donated cells into the host tissue, electrical coupling.
18. A series of conclusive lessons will introduce students to practical issues that need to be addressed when experimenting the use of stem cells for tissue healing. Specifically, cell growth, cell selection methods, and methods for the identification of donated cells into host tissues will be explained and shown in the laboratory.

1. Diagnostic and experimental technologies, nanotechnologies
1. Epifluorescence and confocal microscopy,
2. Fluorescence-based assays,
3. Fluorescence image acquisition and quantification,
4. FRET, FRAP, FLIM, FLIP,
5. Two-photon microscopy,
6. Electron microscopy: TEM and SEM,
7. Scanning probe microscopy: tunnel microscopy and atomic force microscopy,
8. Flow cytometry and flow-cytometry based assays,
9. Separation techniques: fluorescence –activated and immunomagnetic cell sorting,
10. In vivo imaging techniques,
11. Cell cultures,
12. Liquid biopsy: CTC, exosomes and cfDNA,
13. Nanotechnologies.

1. Applicazioni mediche delle biotecnologie e della biologia artificiale
1. Meccanismi di danno d’organo causati da: inadeguato apporto di ossigeno e nutrienti, sia nella fase calda, che durante l’schemia fredda.
2. Effetto delle lesioni associate alla preservazione di organi ideali e organi marginali.
3.  Metodiche di preservazione d’organo convenzionali.
4. Metodiche di preservazione innovative: macchina di perfusione ipotermica.
5. Metodiche di preservazione innovative: macchina di perfusione normotermica. 

ATTIVITÀ DI APPRENDIMENTO E METODI DIDATTICI PREVISTI

This course will include both taught modules and laboratory practice. This latter will be aimed at showing how the competences acquired by the students need to integrate with specific technical skills to address specific, clinically relevant, questions. 

MODALITÀ DI VERIFICA DELL’APPRENDIMENTO

The exam will be written, constituted part of multiple choice quizzes, part of open questions.

TESTI/BIBLIOGRAFIA

Students will be provided both of copy of the slides employed by the Lecturers, and of review and original papers that are considered of crucial importance.

Those that are interested in further reading, may refer to “Principles of Regenerative Medicine” Author(s): Anthony Atala, MD, Robert Lanza, MD, James A. Thomson, PhD, and Robert M. Nerem, PhD ISBN: 978-0-12-369410-2.