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. 2003 Sep 30;100(Suppl 1):11911–11916. doi: 10.1073/pnas.2034195100

Fig. 5.

Fig. 5.

Neural differentiation of Cyno ES cells in vitro. (A) Phase contrast microscopy of proliferating Cyno1-derived neural precursors at day 1 in vitro (DIV1). (B) Same cluster of precursors shown at DIV9. The total cell number has increased by 5- to 8-fold over a 9-day period. (Inset) One of many mitotic figures. Immunohistochemical analyses after 5 days of neural differentiation in the absence of bFGF and epidermal growth factor and the presence of ascorbic acid revealed positive staining for glial fibrillary acidic protein (GFAP), an astrocytic marker seen in C, and TUJ1, a neuronal marker seen in D. (E) Sequential exposure to sonic hedgehog, FGF8b, and ascorbic acid yielded an average of 25% TUJ1+ neurons coexpressing tyrosine-hydroxylase (TH), a marker for dopamine neurons. (F) HPLC revealing the release of dopamine (DA) and serotonin (Ser).