Development of cerebellar neurons on Matrigel and Laminin substrates

Development of cerebellar neurons on Matrigel and Laminin substrates

Nath, S.

Apachitei, I. (mentor)

Mechanical, Maritime and Materials Engineering

Biomechanical Engineering

2016-07-21

Spinocerebellar ataxia (SCA) is a neurodegenerative disease of the cerebellum leading to gradual loss of physical control while maintaining full mental capacity. With no cure yet available, patients undergo therapeutic rehabilitation for alleviating some of the SCA symptoms such as poor coordination of gait, eye, speech and hand movements. Understanding biology of the disease, especially of the cerebellar neurons involved, would help in finding treatments for SCA. One of the challenges of this research is related to the efficient differentiation of pluripotent stem cells into mature cerebellar neurons. Tissue engineering approaches involving novel biomaterials platforms may help in directing differentiation of pluripotent cells. Although researchers are able to generate the cerebellar progenitor cells (CPCs) from mouse embryonic stem cells (mESCs), it is still unclear how these neurons develop and mature on various biomaterial platform. In this study, the effects of substrate and dynamic culture conditions on the in vitro differentiation, dendritogenesis and functionality of mouse embryonic stem cells (mESC)-derived cerebellar progenitor cells have been examined both qualitatively and quantitatively over a culture period of 15 days. Differentiation and dendritogenesis of the CPCs were assessed by immunofluorescence at specific time points over the culture period, while the functionality of the differentiated cerebellar neurons has been assessed by recording electrophysiological activity. Viable and functional neurons were found on both Laminin and Matrigel substrate. However, Matrigel as a substrate enhanced cerebellar neuronal differentiation and resulted in extensive dendritogenesis as compared to Laminin substrate. Differentiated neurons revealed varying spontaneous intrinsic firing activity. This study provides a good foundation for future research on the optimisation of the microenvironment for in vitro differentiation and maturation of the CPCs which can be further help in developing effective treatment for SCA.

Cerebellar progenitor cells (CPCs)
in vitro culture
Matrigel
Laminin
Differentiation
Dendritogenesis
Intrinsic firing activity
Neurons
Glial cells

http://resolver.tudelft.nl/uuid:abe25979-4187-4c30-882b-a72288853184

2021-07-21

Student theses

master thesis

(c) 2016 Nath, S.