%0 Journal Article
%T Modeling Parkinson¡¯s disease in midbrain-like organoids
%A Alessandra Zanon
%A Andrew A. Hicks
%A Anna S. Monzel
%A Emanuel Berger
%A Florian Wegner
%A Jared L. Sterneckert
%A Jens C. Schwamborn
%A Jessica Bellmann
%A Kathrin Hemmer
%A Lisa M. Smits
%A Lydia Reinhardt
%A Marc Ehrlich
%A Marcelo D. Rosato-Siri
%A Michael Glatza
%A Nancy Stanslowsky
%A Norman Kalmbach
%A Paul M. Antony
%A Peter Reinhardt
%A Sarah M. Nicklas
%A Silvia Bolognin
%A Xiaobing Qing
%J Archive of "NPJ Parkinson's Disease".
%D 2019
%R 10.1038/s41531-019-0078-4
%X Generation of midbrain-specific organoids. a Illustration of the conditions used to differentiate spherical midbrain floor plate neural progenitor cells (mfNPCs) into human midbrain-specific organoids (hMOs). mfNPCs are stably expandable up to passage 15; day X indicates the start of dopaminergic differentiation. SB£¿=£¿SB-431542, LDN£¿=£¿LDN-193189, SAG£¿=£¿sonic hedgehog agonist, BD£¿=£¿brain-derived neurotrophic factor, GD£¿=£¿glial cell-derived neurotrophic factor. b Maximum intensity projection of midbrain dopaminergic neuron (mDAN) markers TH and FOXA2 in 70-day-old hMO sections. Scale bar is 100£¿¦Ìm. hMOs derived from mfNPC lines H1¨C3 (image shows line H3). c Immunohistological stainings of mDAN markers TH, FOXA2, EN1, and LMX1A in the center of hMO sections. Scale bar is 50£¿¦Ìm. hMOs derived from mfNPC lines H1¨C3 (images shows line H2). d Bright-field image showing neurons obtained from three-dimensional (3D) culture of line H4 where a recorded cell is attached to the patch-clamp pipette tip (¡Á10 magnification). Scale bar is 20£¿¦Ìm. Representative current-clamp trace of the pace-making firing activity obtained from the recorded mDAN is shown. Experiment conducted with mfNPC lines H4, P2-GC, and P3-GC, in total 26 cells have been recorded, 12 of these showed a similar pattern; representative images and traces of line H4. e Representative immunohistological stainings of mDAN markers FOXA2, TH, dopamine decarboxylase (DDC), and dopamine transporter (DAT) at the edge of an organoid. Scale bar is 20£¿¦Ìm. hMOs were derived from mfNPC lines H1¨C4 (images show lines H3 and H1). f Appearance of dark granules in hMOs at day 255, derived from mfNPC line H1. Scale bar is 500£¿¦Ìm. Fontana-Masson staining reveals neuromelanin-like granules at the edge of the organoid section after 100 days in culture, derived from mfNPCs line H4. Scale bar is 50£¿¦Ìm. g Representative immunohistological staining of spherical maintenance mfNPCs and 70-day hMO sections (50£¿¦Ìm thickness, taken at the edge of a section) for TH and neurotransmitter dopamine (DA). Scale bar is 50£¿¦Ìm. hMOs derived from mfNPC lines H1¨C3 (images show line H3). h Quantitative analysis of DA extracted from the supernatant of mfNPCs and hMOs, *p <£¿0.05. The two time points analyzed in all the experiments were 35 and 70 days of differentiation. Data are presented as mean£¿¡À£¿SEM (mfNPC line H3, 4 different passages, n£¿=£¿4
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6450999/