Differentiation of monkey embryonic stem cells into neural lineages

Hung Chih Kuo, K. Y.Francis Pau, Richard R. Yeoman, Shoukhrat M. Mitalipov, Hideyuki Okano, Don P. Wolf

    Research output: Contribution to journalArticle

    51 Scopus citations

    Abstract

    Embryonic stem (ES) cells are self-renewing, pluripotent, and capable of differentiating into all of the cell types found in the adult body. Therefore, they have the potential to replace degenerated or damaged cells, including those in the central nervous system. For ES cell-based therapy to become a clinical reality, translational research involving nonhuman primates is essential. Here, we report monkey ES cell differentiation into embryoid bodies (EBs), neural progenitor cells (NPCs), and committed neural phenotypes. The ES cells were aggregated in hanging drops to form EBs. The EBs were then plated onto adhesive surfaces in a serum-free medium to form NPCs and expanded in serum-free medium containing fibroblast growth factor (FGF)-2 before neural differentiation was induced. Cells were characterized at each step by immunocytochemistry for the presence of specific markers. The majority of cells in complex/cystic EBs expressed antigens (α-fetal protein, cardiac troponin I, and vimentin) representative of all three embryonic germ layers. Greater than 70% of the expanded cell populations expressed antigenic markers (nestin and musashil) for NPCs. After removal of FGF-2, approximately 70% of the NPCs differentiated into neuronal phenotypes expressing either microtubule-associated protein-2C (MAP2C) or neuronal nuclear antigen (NeuN), and approximately 28% differentiated into glial cell types expressing glial fibrillary acidic protein. Small populations of MAP2C/NeuN-positive cells also expressed tyrosine hydroxylase (∼4%) or choline acetyltransferase (∼13%). These results suggest that monkey ES cells spontaneously differentiate into cells of all three germ layers, can be induced and maintained as NPCs, and can be further differentiated into committed neural lineages, including putative neurons and glial cells.

    Original languageEnglish (US)
    Pages (from-to)1727-1735
    Number of pages9
    JournalBiology of reproduction
    Volume68
    Issue number5
    DOIs
    StatePublished - May 1 2003

    Keywords

    • Differentiation
    • Embryonic stem cells
    • Glial cells
    • Monkey
    • Neurons

    ASJC Scopus subject areas

    • Reproductive Medicine
    • Cell Biology

    Fingerprint Dive into the research topics of 'Differentiation of monkey embryonic stem cells into neural lineages'. Together they form a unique fingerprint.

  • Cite this