Directed differentiation of human embryonic stem cells into glial cells
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Human embryonic stem cells are pluripotent cells that have the potential to differentiate into all cell types found in the human body. The Stice Lab has previously derived neural progenitor (hNP) cells, which can be further differentiated into neurons, from human embryonic stem cell lines. Currently, the media used for hNP proliferation comprises of neural basal media, Leukemia Inhibitory Factor, Fibroblast Growth Factor 2, L-glutamine, and an undefined B27 supplement. To better understand the role of B27 on neurons, hNP cells were grown in various concentrations of B27 ranging from 0 to 1XB27. Our results showed 0.1X B27 was sufficient for hNP proliferation. Additionally, RNA analysis data from differentiated neuritis that were proliferated in 0.5X B27 and 0.1X B27 media showed a significantly higher expression of Glial Fibrillary Acidic Protein (GFAP), a marker used to identify glial cells, when compared to our control cells. These observations indicate that hNP cultures proliferating in lower concentrations of B27 differentiate into glial like cells. Immunofluorescent experiments to detect glial proteins levels indicated that the cells did not express GFAP or A2B5, a glial precursor marker, at significant levels. As a result, it was concluded that decreasing the levels of B27 induces transcription of glial genes, however, these genes are not translated into proteins. Glial cells provide support and nutrition for neurons in the central nervous system. Defining a uniform culturing condition for these cells could help understand and cure glial-degenerative diseases like Multiple Sclerosis, Alzheimer's, and Alexander's Disease.