B2195 - Stem Cells

Learning outcomes

At the end of the course the student has basic knowledge of embryonic, somatic stem cells and induced pluripotent stem cells biology, including cell cycle, cell division and differentiation mechanisms, the genetic determinants of stemness, the role of the in situ niche. Has the ability to critically evaluate literature and protocols for the isolation and maintenance of totipotency and differentiation of embryonic stem cells; somatic cells reprogramming; isolation and maintenance of hematopoietic, mesenchymal, epithelial, and neural stem cells from adult tissues. Has the basic knowledge for omics technologies applied to stem cells, including study design and limits of omics application in the field. Knows the technical bases for stem cell use in regenerative medicine and cell therapies, principles of genetic engineering/gene editing for stem cells application in animal transgenesis (from pluripotent stem cells to genetically engineered offspring), reproductive and therapeutic cloning, organoids and organ-on-chip technologies. Has the ability to search and critically evaluate stem cell banks for research purposes.

Course contents

Module 1, Stem Cell, Laura Calzà

Definitions of toti-, multipotentiality, differentiation, terminal differentiation, de-differentiation; embryonic and adult somatic stem cells. Timeline of major discoveries (4 hours)

Adult stem cells: locations, the vascular "niche", cellular and acellular constituents, normal niche and pathological niche. Asymmetric division: cell polarity and role of the niche. Role of Numb and Notch. Omics in the study of somatic stem cells (4 hours)

Skin stem cells. Holoclones, meroclones and paraclones. Role in skin homeostasis and wound repair (2 hours)

Intestinal stem cells. The niche in the intestinal crypt; timing of renewal of different cell types, mechanisms of migration from crypt to villus. Signaling Wnt-beta-catenin; signaling Ephrin. role of Paneth cells (2 hours)

Hematopoietic stem cells: niche composition and stem cell regulation (angiopoietic, Ca++, Shh) (2 hours)

Mesenchymal stem cells: in vivo and in vitro characteristics, adult tissues of origin. Spontaneous and induced differentiation. Paracrine properties (2 hours)

Stem cells from placental and related tissues (amnion, umbilical cord, umbilical cord blood, amniotic fluid, placenta). Cord banking: technical aspects, regulations, and impact for regenerative medicine (2 hours)

Neural stem cells. The system of "neurospheres". Neurogenic regions in the adult Central Nervous System; "numbers", role and regulation of neurogenesis in adulthood. Integration of new neurons into pre-existing circuits: olfactory bulb and dentate gyrus of the hippocampus. Neurogenesis in pathological brains (2 hours)

Stem cells, cell therapies and tissue engineering: brief notes and the role of big data (3 hours)

Take home message: the complexity theory applied to the Life Sciences (1 hour)

 

Module 2 Introduction

The Importance of Stem Cells in Translational Medicine (1 hour)

Embryonic Stem Cells as a Disease Model

Definitions and Key Concepts.

Cell Cycle Regulation (Duration, Characteristics of Different Phases, R-point).

Murine Embryonic Stem Cells.

Isolation, Culture, and Validation Techniques.

Role of Embryonic Stem Cells in Cell Therapy (Brief Overview of Safety and Ethics).

Embryonic Stem Cell Registries and Banks.

(2 hours)

Induced Pluripotent Stem Cells

Definitions and Key Concepts.

Genes and Markers of Pluripotency.

Transcription Factors and Signaling.

Reprogramming Strategies.

Epigenetic Regulation and Cellular Competence.

Culture and Validation Techniques.

Pluripotent Stem Cells in Regenerative Medicine.

Applications in Biomedical Research and Cell Therapy.

Direct Reprogramming.

(4 hours)

Organoids

3D Cultures and Organoids: Characteristics and Definitions.

Methods for Generating and Characterizing Organoids.

The Example of Brain Organoids.

Assembloids: Characteristics and Definitions.

Limitations.

Organ-on-a-Chip.

Applications in Research as Disease Models.

(4 hours)

Genetic Engineering and Stem Cells

Definitions and Key Concepts.

The CRISPR/Cas9 System and Its Components.

DNA Repair Mechanisms in Genetic Engineering.

Variants of the CRISPR/Cas9 System.

Limitations.

Applications of Genetic Engineering in Stem Cells and Therapeutic Approaches.

Ethical Considerations.

(4 hours)

Somatic and Therapeutic Cloning

Definitions and Key Concepts.

Principles and Methods of Somatic Cloning.

Applications of Somatic Cloning in Animal Models.

Limitations.

Therapeutic Cloning: Applications and Limitations.

(3 hours)

Omic Technologies and Stem Cells

Definitions and Key Concepts.

Single-Cell Approach.

Applications in Stem Cells and Therapeutic Approaches.

(3 hours)

Journal Club

Research and Selection of Scientific Articles from Online Databases.

Functional Approach to Scientific Literature.

Methods for Preparing a Scientific Presentation.

(Part 1; 1 hour)

Journal Club

Students will be asked to select articles of interest related to the topics covered in the module and produce a scientific presentation as a starting point for class discussion.

(Part 2; Group Work; 2 hours)

Readings/Bibliography

Stem cells, Gian Paolo Bagnara, Laura Bonsi, Francesco Alviano, Esculapio, 2020

Stem Cells Scientific Facts and Fiction, Christine L. Mummery, Anja van de Stolpe, Bernard Roelen, Hans Clevers, Elesevier, 2021

Teaching methods

Frontal lectures, group discussions, journal club.

Students are hereby informed that, due to recent University directives, it will not be possible to attend the course remotely (e.g., through recordings), except in specific cases that must be duly documented and approved by the University prior to the start of classes.

Assessment methods

Brief PowerPoint presentation (10 minutes) on a topic of the Student's choice from those covered in class or agreed upon with the Instructor, followed by discussion; question at the Instructor's discretion on the remaining portion of the program covered in class. The exam is intended to be joint for the two Modules of the course.

Teaching tools

The ppt presentations used in class will be available on the dedicated spaces.

Office hours

See the website of Laura Calzà

See the website of Corinne Quadalti