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 (4 hours)

 

Module 2, Stem Cell Technologies and Applications,Corinne Quadalti

The role of embryonic stem cells in disease modelling. Cell cycle regulation: cycle length, R-point and S-G1 transition. Methods for isolation and culture of ESCs. Role of ESCs in cell therapy. The example of murine embryonic stem cells. Human embryonic stem cells repositories and biobanks. (3 hours)

Pluripotent and induced pluripotent stem cells. Genes and markers of pluripotency. Transcription factors and signalling in pluripotency: Oct4, Sox2, Nanog, LIF, BMP. Epigenetic regulation. Pluripotent stem cells in regenerative medicine. Reprogramming strategies for the generation of iPSCs. Competence in cellular reprogramming. PSCs and iPSCs in disease modelling and cell therapy. Isolation techniques and culture method. (4 ore)

3D coltures: organoids and assemploids generation and characterization methods. Organ-on-a-chip. Application of 3D colture in medical research as disease models. (4 hours)

Genetic engineering in stem and pluripotent stem cells for the generation of disease models. Overview of protocols, available tools, main applications and limitations. (4 hours)

Somatic and therapeutic cloning. Overview of SCNT protocols. Application of SCNT for the generation of animal models and relative limitations. Application examples. (3 hours)

“Omic” technologies applied to stem cells and 3D coltures: how to plan an experiment and possible limitations. Application examples. (4 hours)

Intrgration of “omic” sciences with computational biology in stem cells and 3D systems research. Analysis of reference datasets. (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

Lectures, group discussions

Assessment methods

Short ppt presentations of a topic covered in class or agreed with the teacher, and related discussion

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