Stem Cells 2

11 Maggio 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

Lesson 1. Molecular logic of cortical development. Evolution of pallial development by 2D cultures: Human and primate PSCs can replicate cortical development in culture. PSC-derived cortical progenitors from different species expand to different degrees. Clonal analysis reveals marked difference in neurogenesis output over time. Species-specific timing differences in neurogenesis are regulated cell autonomously.

15 Maggio 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

Lesson 2. Notch-Delta signaling in corticogenesis: selection of differentiating cells by different levels of delta-like 1 among neural precursor cells in the developing mouse telencephalon. Pallial evolution: Human-Specific NOTCH2NL Genes Expand Cortical Neurogenesis through Delta/Notch Regulation.

25 Maggio 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

Lesson 3. Review on different protocols to obtain and use cerebral organoids for base and transaltional research. Cerebral organoids model human brain development and microcephaly.

29 Maggio 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

Lesson 4. Pallial evolution: cortical organoid approach. Establishing Cerebral Organoids as Models of Human-Specific Brain Evolution. Primate cell fusion disentangles gene regulatory divergence in neurodevelopment

08 Giu 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

Cortical organoids can be used to model brain disorders due to defective cortical layering. To overcome the diffusion limit a slicing method can be used to prevent interior cell death and sustain the generation of thick supragranular cortical layers. Using this approach, the role of DISC1 mutation in affecting the cortical layering of patients with cognitive impairment was dissected. Adherent cultures can be successfully employed to investigate the role of single gene mutations in the malfunctioning of cortical neural networks of patients. Sparse co-culture for connectivity assays showed that SHANK2 gene mutation causes increases in dendrite length, dendrite complexity, synapse number, and frequency of spontaneous excitatory postsynaptic currents.

12 Giu 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

Lesson 6. Human pluripotent stem cells (PSCs) are a source of cells for applications in regenerative medicine. Directed differentiation of hiPSCs into specialized cells such as midbrain dopamine (DA) neurons has been achieved. 25 and can be maintained in vitro for several months. Extensive molecular profiling, biochemical and electrophysiological data define developmental progression and confirm identity of hiPSC-derived midbrain DA neurons. Long-term engraftment in 6-hydroxy-dopamine-lesioned mice and rats demonstrates robust survival of midbrain DA neurons derived from hiPSCs, complete restoration of amphetamine-induced rotation behaviour and improvements in tests of forelimb use and akinesia.

15 Giu 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

Lesson 7. Assembly of functionally integrated human forebrain spheroids. Generation of three-dimensional spheroids from human pluripotent stem cells that resemble either the dorsal or ventral forebrain and contain cortical glutamatergic or GABAergic neurons allows the modeling of embryonic interneuron saltatory migration. Using this approachl, Timothy syndrome was modeled using patient-derived ips cells and abnormal migratory saltations of interneurons was displaied.

26 Giu 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

Lesson 8. Human heart-forming organoids recapitulate early heart and foregut development. Complex, highly structured, three-dimensional heart-forming organoids (HFOs) were generated by embedding human pluripotent stem cell aggregates in Matrigel, followed by directed cardiac differentiation via biphasic WNT pathway modulation with small molecules. The architecture of HFOs closely resembles aspects of early native heart anlagen before heart tube formation. This method allowed to study genetic defects in vitro by demonstrating that NKX2.5-knockout HFOs show a phenotype reminiscent of cardiac malformations previously observed in transgenic mice.

03 Lug 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

lesson 9. Generation and characterization of hair-bearing skin organoids from human pluripotent stem cells. The differentiation of pluripotent stem cells to surface ectoderm and cranial neural crest cells upon controlled conditons gives rise to the epidermis and dermis, respectively, in organoids. The exact timing of Wnt signaling is crucial to induce self-organizing organoid containing stratified skin layers, pigmented hair follicles, sebaceous glands, Merkel cells and sensory neurons,

18 Lug 2023 (2h 00m)

Federico Cremisi - Corso (attività didattica) - In presenza

Lesson10. Xenotransplanted brain organoids as in vivo platform for studying human microglia (hMGs); hMGs gain human-specific transcriptomic signatures and assume in-vivo-like identities. hMGs engage in surveilling the human brain environment and react to perturbations. A patient-derived model reveals a brain-environment-induced immune response in autism