1. What is a stem cell? Different definitions for different purposes. Pluripotent stem cell; in vivo stem cell; in vitro clonogenic cell; transplantable cell; cell expressing certain markers; label retaining cell. 2. Early mouse and human development. Gametogenesis, fertilization, cleavage, ICM-TE formation, epiblast-primitive endoderm, extraembryonic ectoderm, X-inactivation, implantation, placenta, sex determination. Specifics of human development. 3. Pluripotent stem cells. Mouse embryonic stem cells, human embryonic stem cells, naive and primed states, induced pluripotent stem cells. Somatic cell nuclear transfer. Ethical issues surrounding embryonic stem cells. 4. Body plan formation and organogenesis. Developmental commitment. Hierarchical nature of development. Major signalling systems (Wnt, FGF etc). Key genes characterising developmental states (Oct4, Sox2, Bra, Cdx2, p63, Hox). Development of CNS, epidermis, myogenesis, cardiogenesis, blood and blood vessels, intestine, pancreas. Do postnatal pluripotent stem cells exist? 5. Cell differentiation. Organs, tissues and cell types. Histological classification. Lateral inhibition: Notch and bHLH factors. Neurogenesis, differentiation in epithelia. Transdifferentiation. Design of differentiation protocols for pluripotent stem cells. 6. Characterizing cells. Sections and wholemounts, laser dissection, RNAseq, in situ hybridization, immunostaining, flow cytometry. The cell cycle. Intrinsic markers. DNA synthesis markers. Cell death markers. 7. Genetic modification and labelling of cell lineages. Introducing genes to cells, homologous recombination in mice, CRISPR-Cas. Principles of clonal analysis; passive lineage labels, genetic labels. CreER system in mice. 8. Tissue culture and tissue engineering. Culture media, gases, serum and growth factors, defined media, GMP cultivation, organ culture, suspension culture. Scaffolds, 3D cultures, 3D printing of matrices and cells. Recellularization of decellularized tissues and organs. 9. Stem cells in the body. Epidermis, neural stem cells, muscle satellite cells, haematopoiesis, spermatogenesis, mesenchymal stem cells, intestinal epithelium. Symmetric and asymmetric division. Mechanisms of asymmetric division in C.elegans, Drosophila, mouse. Regeneration in planaria and the amphibian limb. 10. Cancer and cancer stem cells. Multi-hit mutagenesis; clonal analysis of tumours; definition of cancer stem cells; implications for therapy.
