Stem Cells 2

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Stem Cells 1


The protocols underlying the in vitro neuralization of pluripotent cells (ESCs, iPSc) and their logical for using intracellular signaling molecules in relation to the known developmental processes of the early embryo.

Molecular control of neural induction. The signaling of Wnt and that of Tgf-beta in the induction of the organizer's BMP inhibitors. The effect of Wnt in the induction of key neuralization genes in presumptive neural ectoderna. Induction of the regional specificity of the nervous system. Function of the node and AVE in the postero-anterior polarization of the mammalian nervous system. Evolutionary conservation of neuralization and antero-posterior patterning signals in vertebrates. The regulation of dorso-ventral patterning in the neural tube: the example of the spinal cord. The gradients of BMP and SHH are translated into segments of positional identity thanks to the induction of transcription factors such as Olig2 and nkx2.2, which are endowed with different activation sensitivities and able to co-repress their own expression.

Production of human neurons with dorsal and ventral positional identity of various CNS and SNP regions through the temporally targeted use of agonists and antagonists of the Wnt, BMP, Shh, Tgfb, FGF, RA signaling pathways.

In vitro modeling of the embryonic development of the retina, cerebral cortex, hippocampus, spinal cord, neural crest derivatives, for the discovery of new molecular mechanisms of cell identity specification, diagnosis and treatment of genetic and neurodegenerative diseases.

Role of Notch-Delta in the control of proliferation and neurogenesis of the mammalian cerebral cortex: in vivo and in vitro studies. Mechanisms of regulation of the evolutionary expansion of the cerebral cortex. The developmental logic of the mammalian cerebral cortex: acquisition of area and layer identity. The paracrine signaling centers and the master regulatory genes of area and layer identity. The Bolean logic model of successive specifications of cortical cellular identity.

Use of reprogrammed human cells hiPSCs to model development and pathologies of the cerebral cortex. Cell cultures in adhesion and cerebral organoids can faithfully reproduce the stratification process of the human cortex in vitro.

Use of cerebral organoids to study the effects of the DISC1 gene mutation, responsible for the onset of autism spectrum disorders, in the stratification of the cortex. 2D and 3D cultures of cortical neurons derived from iPSCs cells of chimpanzees, macaques or humans show the presence of a genetic program intrinsic to the neural progenitor capable of directing and prolonging cortical neurogenesis with the timing and modalities of the brain development of the three species. 2D cultures of cortical neurons derived from hiPSCs from human patients for in vitro modeling of human cortical neural networks. Study of the structural and functional consequences of the SHANK2 gene mutation, which causes autism spectrum disorders.

Interface with mouse and human neuronal networks in culture for the study of spontaneous and induced electrical activity. The methodology of Multiple Electrodes Arrays (MEA) and calcium sensors for the direct and indirect recording of the potentials of neuronal networks.

Obiettivi formativi

Acquisition of specific knowledge on in vitro models of development and function of brain regions using reprogrammed cells

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