Inhibition of DNA methyltransferases and histone deacetylases induces astrocytic differentiation of neural progenitors

• Published in: Stem cell research 2013-07, Vol.11 (1), p.574-586
• Main Authors: Majumder, Anirban, Dhara, Sujoy K., Swetenburg, Raymond, Mithani, Miloni, Cao, Kaixiang, Medrzycki, Magdalena, Fan, Yuhong, Stice, Steven L.
• Format: Article
• Language: English
• Subjects: Astrocytes - cytology; Astrocytes - drug effects; Astrocytes - enzymology; Astrocytes - metabolism; Azacitidine - pharmacology; Cell Differentiation - drug effects; DNA - metabolism; DNA Modification Methylases - antagonists & inhibitors; DNA Modification Methylases - metabolism; Enzyme Inhibitors - pharmacology; Histone Deacetylase Inhibitors - pharmacology; Histone Deacetylases - metabolism; Humans; Hydroxamic Acids - pharmacology; Immunohistochemistry; Neural Stem Cells - cytology; Neural Stem Cells - drug effects; Neural Stem Cells - enzymology; Neurogenesis - drug effects; Neurons - cytology; Neurons - drug effects; Neurons - enzymology; Neurons - metabolism
• ISSN: 1873-5061; 1876-7753
• DOI: 10.1016/j.scr.2013.03.003

Understanding how to specify rapid differentiation of human neural progenitor towards enriched non-transformed human astrocyte progenitors will provide a critical cell source to further our understanding of how astrocytes play a pivotal role in neural function and development. Human neural progenitors derived from pluripotent embryonic stem cells and propagated in adherent serum-free cultures provide a fate restricted renewable source for quick production of neural cells; however, such cells are highly refractive to astrocytogenesis and show a strong neurogenic bias, similar to neural progenitors from the early embryonic central nervous system (CNS). We found that several astrocytic genes are hypermethylated in such progenitors potentially preventing generation of astrocytes and leading to the proneuronal fate of these progenitors. However, epigenetic modification by Azacytidine (Aza-C) and Trichostatin A (TSA), with concomitant signaling from BMP2 and LIF in neural progenitor cultures shifts this bias, leading to expression of astrocytic markers as early as 5days of differentiation, with near complete suppression of neuronal differentiation. The resultant cells express major astrocytic markers, are amenable to co-culture with neurons, can be propagated as astrocyte progenitors and are cryopreservable. Although previous reports have generated astrocytes from pluripotent cells, the differentiation required extensive culture or selection based on cell surface antigens. The development of a label free and rapid differentiation process will expedite future derivation of astrocytes from various sources pluripotent cells including, but not limited to, human astrocytes associated with various neurological diseases. •We describe rapid astrocytic differentiation of human neural progenitors (hNP).•hNP cells are epigenetically biased towards neuronal fates.•We suggest hypermethylated astrocytic gene promoters as the mechanism for this bias.•We overcome the bias using small molecules for astrocyte differentiation.