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Thesis Project Form

Title (tentative): Chitosan as adhesion factor: from promotion to polarization of neurite growth of neurons derived from human induced pluripotent stem cells.

Thesis advisor(s): Pastorino Laura E-mail:
Address: Via Opera Pia 13 Phone: (+39) 010 33 56547
Description

Motivation and application domain
The use of chitosan has been already investigated as an alternative cell adhesion factor for in vitro preparation of two dimensional primary neuronal culture. The investigation of the ability of chitosan to promote neuronal differentiation of neurons derived from human pluripotent stem cells is still unknown. This process is fundamental for the function of the nervous system since it specialized for transmitting signals in a polarized manner. Neurite branching affects how single neurons integrate synaptic inputs and how they communicate as networks. Neuronal morphology is a complex phenomenon to study due to the wide range of metrics which may be quantified.

General objectives and main activities
Aim of this thesis is to validate the ability of chitosan to support neuronal adhesion, differentiation and networks development of neurons derived from human induced pluripotent stem cells. The morphological characterization will be solved by the Bonfire program, a series of custom scripts written in MATLAB (MathWorks). Neuronal morphology will be digitized in three stages based on the initial images that will involve the use of the semi-automated tools available through the NeuronJ plugin to ImageJ (NIH, Bethesda, MD) to acquire spatial information about the position of neuritic segments in space relative to the rest of the cell and NeuronStudio to define structural information about the connectivity between neurites. Moreover, a functional characterization will be carry out by synaptic count. The experimental activity will be aimed at the in vitro tests onto different types of chitosan followed by the observation and characterization of the induced neurons during the different development stages.

Training Objectives (technical/analytical tools, experimental methodologies)

Place(s) where the thesis work will be carried out: DIBRIS, University of Genova

Additional information

Maximum number of students: 1