stemdiff astrocyte differentiation kit  (STEMCELL Technologies Inc)

Journal: Journal of Neurochemistry

Article Title: Altered metabolic function induced by Aβ‐oligomers and PSEN1 mutations in iPSC ‐derived astrocytes

doi: 10.1111/jnc.16267

Figure Lengend Snippet: Characterisation of ‘healthy’ iPSC‐derived astrocytes. (a) Representative images showing differentiation and ICC staining of iPSC‐derived astrocytic cells at Day 45+. The astrocytes were differentiated from ‘healthy’ control NPCs for >40 days using astrocytes differentiation and maturation protocols. Top) Phase contrast images of control astrocytes on Days 2, 10 and 40 in culture post differentiation. Bottom) Cells were stained using immunofluorescent antibodies for astrocytic markers ALDH1L1 (left, green), S100β (middle, green) and GFAP (right, red), nuclei were counterstained in each image with DAPI (blue). Scale bars: 100 μM. (b) Cellular glycogen content of control astrocytes following exposure to hypoglycaemic conditions and treatment with DAB over 60 and 120 min. (c) Glycogen content of cells treated with dbcAMP an isoproterenol. (d) Glycogen contents of cells treated with Glutamate, oubain, glutamate and ouabain, TBOA and glutamate and TBOA for 60, 180 and 360 min. Results are expressed as ug/mg protein ± SD, n = 3 p < 0.05 (*), p < 0.01 (**), p < 0.001 (***). For DAB (b), a two‐way analysis of variance (ANOVA) was performed follow by Sidaks post‐test. Comparisons between treatments (c) were performed using analysis of variance (ANOVA) followed by Dunnet's post‐test. Each replicate or ‘ n ’ represents an independent culture preparation and is displayed as an individual data point.

Article Snippet: Cells were plated at 5 × 10 4 /cm 2 density and maintained in a 37°C, 5% CO 2 / 95% air atmosphere with a total medium exchange every other day, through two subsequent passages, before switching to astrocyte maturation medium (STEMdiff tm Astrocyte maturation kit #100‐0016, StemCell Technologies, Cambridge, UK).

Techniques: Derivative Assay, Staining, Control