Journal: Tissue Engineering. Part A

Article Title: Peptide-Based Scaffolds for the Culture and Transplantation of Human Dopaminergic Neurons

doi: 10.1089/ten.tea.2019.0094

Figure Lengend Snippet: Protocol for differentiation and encapsulation of DA neurons. (A) Protocol for expansion of DA neural progenitors and final differentiation of DA neurons. (B) Representative phase-contrast images of DOPA1, DOPA2, and DOPA3 cells. Scale bar: 100 μm. (C) Timeline for encapsulation within SAPNS. DA, dopaminergic; SAPNS, self-assembling peptide nanofiber scaffolds. Color images are available online.

Article Snippet: Twenty-four hours later, neurospheres were plated onto a six-well tissue culture polystyrene (TCPS) dish (Eppendorf) ( Supplementary Figure S2 ) coated with 5 μg/mL laminin (MilliporeSigma), in DOPA2 medium (base medium supplemented with 0.5 μM SAG [9128694; Biogems]), 5 μM Y-27632, and 10 ng/mL of the following growth factors (PeproTech): stromal cell-derived factor 1-alpha (SDF-1α); Pleiotrophin; hepatocyte growth factor (HGF); insulin-like growth factor-2 (IGF-2); vascular endothelial growth factor-D (VEGF-D); secreted frizzled-related protein 1 (SFRP-1), fibroblast growth factor-8 (FGF-8); and 20 ng/mL cerebral dopamine neurotrophic factor (CDNF).

Techniques:

Journal: Tissue Engineering. Part A

Article Title: Peptide-Based Scaffolds for the Culture and Transplantation of Human Dopaminergic Neurons

doi: 10.1089/ten.tea.2019.0094

Figure Lengend Snippet: Characterization of DA neural progenitors and DA neurons. (A) Immunocytochemical staining of the following markers: Nestin: neural stem/progenitor, Tuj1 (β-III tubulin): neuron, MAP2: mature neuron, TH: DA neuron, Ki-67: proliferative cells, Synapsin: synaptic vesicle, FOXA2: midbrain DA neuron, PITX3: midbrain DA neuron, En-1 (Engrailed-1): midbrain DA neuron, DAPI: nucleus. Scale bar: 50 μm. (B) qRT-PCR analysis comparing gene expression in 2D and 3D (SAPNS-encapsulated) DA neurons at day 22 of differentiation. Data are presented as log2 of the fold change in expression relative to DOPA2 neural progenitors, shown in a bar graph (left) and heat map (right). Error bars represent standard deviation. p > 0.05, Student's unpaired t-test. 2D, two-dimensional; 3D, three-dimensional; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; TH, tyrosine hydroxylase. Color images are available online.

Article Snippet: Twenty-four hours later, neurospheres were plated onto a six-well tissue culture polystyrene (TCPS) dish (Eppendorf) ( Supplementary Figure S2 ) coated with 5 μg/mL laminin (MilliporeSigma), in DOPA2 medium (base medium supplemented with 0.5 μM SAG [9128694; Biogems]), 5 μM Y-27632, and 10 ng/mL of the following growth factors (PeproTech): stromal cell-derived factor 1-alpha (SDF-1α); Pleiotrophin; hepatocyte growth factor (HGF); insulin-like growth factor-2 (IGF-2); vascular endothelial growth factor-D (VEGF-D); secreted frizzled-related protein 1 (SFRP-1), fibroblast growth factor-8 (FGF-8); and 20 ng/mL cerebral dopamine neurotrophic factor (CDNF).

Techniques: Staining, Quantitative RT-PCR, Expressing, Standard Deviation, Reverse Transcription Polymerase Chain Reaction