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Conversion of murine embryonic stem cells to neural precursor cells, astrocytes and neurons by differentiation and corticogenesis and its characterization
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Conversion of murine embryonic stem cells to neural precursor cells, astrocytes and neurons by differentiation and corticogenesis and its characterization

Hansini Upadhyay
Master of Science (M.S.), Drexel University
01 Jun 2015
DOI:
https://doi.org/10.17918/etd-6507
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Abstract

Embryonic stem cells--Research MATLAB Biomedical Engineering Cell Differentiation
ES cells have been used for many years now to generate specific cell lines of neural cells and they have also emerged as a promising approach in developmental neurobiology, by providing models for neural development. Several scientists have tried differentiating ES cells into neurons and have often described this process to be cumbersome. However, there are alternative methods available to the differentiation protocol which is shorter in time period than the traditional differentiation methods. This method is called genetic reprogramming. My aim was to compare traditionally differentiated cells to primary cells and characterize them by observing the difference in morphology and their ability to express differentiated cell markers and quantify my results by counting the number of cells expressing the marker using MATLAB. I chose to focus first on differentiation of murine ES cells to neural progenitor cells (NPC's) as they have the ability to differentiate into many terminally differentiated neural cell types. By optimizing coating and culture conditions, I have established an enriched population of neural progenitor cells. As mentioned, these cells could then be used to generate a variety of neural cell-types; I chose to optimize differentiation of NPCs down the glial lineage. ES cells also yield cortical neurons when they undergo a process called Corticogenesis. Corticogenesis is the formation of cortex and this is obtained by using a chemically defined medium in the absence of morphogen but in the presence of a sonic hedgehog inhibitor, Cyclopamine. It causes the cells to directly shift their fate towards neuronal lineage by blocking the Sonic Hedge Hog (Shh) pathway. During this process, neurogenesis is spotted at day 6 which marks the onset of neural lineage formation. Corticogenesis from ESC's recapitulates the most important steps of cortical development, leading to the generation of multipotent cortical progenitors that sequentially produce cortical pyramidal neurons in 3 weeks.

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