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Induction of midbrain dopaminergic neurons using extrinsic factors.

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posted on 17.01.2012, 00:28 by Anna Falk, Philipp Koch, Jaideep Kesavan, Yasuhiro Takashima, Julia Ladewig, Michael Alexander, Ole Wiskow, Jignesh Tailor, Matthew Trotter, Steven Pollard, Austin Smith, Oliver Brüstle

IPS cell derived lt-NES cells cultured for 7 days in the presence of sonic hedgehog (Shh) and FGF8 showed nuclear expression of Lmx1a and FoxA2 (A). (B) With prolonged (14 days) exposure to Shh and FGF8, the number of cells expressing FoxA2 gradually increased. Shh/FGF8-treated cells were also positive for the midbrain marker En1, whereas neither FoxA2 nor Lmx1a were expressed in control populations cultured in the absence of Shh/FGF8. (C–D) Growth factor removal from lt-NES cell lines grown in Shh/FGF8 for 14 days induced differentiation and appearance of cell clusters positive for FoxA2 and tyrosine hydroxylase (TH), with beta III-tubulin-positive neurons co-expressing Nurr1. In contrast, control cultures propagated in FGF2/EGF remained TH-negative. Abbreviations in upper right corners of the immunofluorescence photomicrographs denote lt-NES cell lines used. Scale bars: 50 µm.