Nanoparticle-mediated conversion of primary human astrocytes into neurons and oligodendrocytes.
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ABSTRACT:
SUBMITTER: Li X
PROVIDER: S-EPMC4922536 | biostudies-literature | 2016 Jun
REPOSITORIES: biostudies-literature
Publications
Nanoparticle-mediated conversion of primary human astrocytes into neurons and oligodendrocytes.
Li Xiaowei X Kozielski Kristen K Cheng Yu-Hao YH Liu Huanhuan H Zamboni Camila Gadens CG Green Jordan J Mao Hai-Quan HQ
Biomaterials science 20160601 7
Central nervous system (CNS) diseases and injuries are accompanied by reactive gliosis and scarring involving the activation and proliferation of astrocytes to form hypertrophic and dense structures, which present a significant barrier to neural regeneration. Engineering astrocytes to functional neurons or oligodendrocytes may constitute a novel therapeutic strategy for CNS diseases and injuries. Such direct cellular programming has been successfully demonstrated using viral vectors via the tran ...[more]
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The nonadherent cells from the last O4 dish were harvested by centrifugation, and the cells were resuspended in panning buffer containing GalC, MOG, and O1 supernatant and incubated for 15 minutes at room temperature. The cell suspension was washed and then resuspended in panning buffer containing 20 μg donkey anti-mouse APC for 15 minutes. The cells were washed and resuspended in panning buffer containing propidium iodide (PI). EGFP+ astrocytes were then purified by fluorescence activated cell sorting (FACS). Dead cells were gated out using high PI staining and forward light scatter. Astrocytes were identified based on high EGFP fluorescence and negative APC fluorescence from indirect immunostaining for OL markers GalC, MOG, and O1. 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