Table 1.
The list of primers used in this study.
Fig 1.
A neuron-laden agarose-laminin hydrogel.
A) A workflow diagram for cell-laden agarose-laminin scaffold preparation B) The morphology of 3D culture in two different platforms; scaffold (top) and scaffold-free (bottom) culture at day 2, 4, and 6 post-seeding. At day 6 of cell culture under scaffold-free platform, SH-SY5Y cells were aggregated and began to attach on the plastic surface (red arrow). Scale bar represents 50 μm. C) The expression of mature neural marker, TUJ1 of scaffold and scaffold-free 3D cultures at day 7. Bar graph represents the mean ± SE (n = 3). n.s: not statistically different.
Fig 2.
An amyloidogenic induction in vitro A) A workflow diagram of the amyloidogenic induction in cell-laden agarose-laminin scaffold. B) A workflow diagram of the cell retrieval from cell-laden agarose-laminin scaffold.
Fig 3.
Small molecule-induced amyloid-β 42 production A) The concentration of total amyloid-β (40+42 isoforms) analyzed by ELISA assay. B) The ratio of total amyloid-β 42 to amyloid-β 40. C) The percentage of two isoforms in total amyloid-β. (n = 5) *: p<0.05. n.s: not statistically different. D) A total protein of 1 μg was blotted on nitrocellulose membrane and probed with actin-β antibody.
Fig 4.
Gene expression analysis A-D) The expression of amyloid-related genes, APP PS1, PS2, and BACE1, respectively upon Aftin-4 treatment for 24 hours. Bar graph represents the mean ± SE (n = 3). **: p<0.01. n.s: not statistically different. E-H) Melt curves analysis of APP, PS1, PS2, and BACE1, respectively from all samples.
Fig 5.
Flow cytometry analysis A) The cell morphology after retrieval and DAPI staining. Scale bars represent 50μm. B) The cellular characteristics; forward scatter (FSC) and side scatter (SSC) of cell derived from 2D culture and D) 3D agarose-laminin scaffold counterpart. C) The histogram of Nestin staining to identify the neural progenitor stage of cells derived from 2D culture and E) agarose-laminin scaffold. F) The histogram of TUJ1 staining to identify the mature neurons derived from 2D culture at DIV3, G) 2D culture at DIV10 and H) 3D agarose-laminin scaffold at DIV10.