microglial differentiation medium  (Corning Life Sciences)

Journal: bioRxiv

Article Title: SARS-CoV-2 infection in hiPSC-derived neurons is cathepsin-dependent and causes accumulation of HIF1ɑ and phosphorylated tau

doi: 10.1101/2024.11.21.624622

Figure Lengend Snippet: A. Representative image of anti-Iba-1 (microglia) and anti-MAP2 (neuron) staining of an untreated neuron-astrocyte-microglia co-culture. B. Same image but the Iba-1 staining has been thresholded with ImageJ into binary format to better display the microglial morphology. C. Similar thresholded image of SARS-CoV-2 infected culture. D Representative image of a SARS-CoV-2 infected neuron (stained with anti-N protein, arrowhead) among Iba-1 positive microglia in triplecultures. E. Mean intensity of AT8 fluorescent signal in untreated, treated but noninfected, and infected neurons at 48 hpi. N = 6 replicate wells. F. and at 72 hpi. N = 3 replicate wells. Scale bars 100 µm. All statistical significances were tested using Kruskal-Wallis test with Dunn’s correction for multiple testing. Only significant results are denoted. *, P ≤ 0.005.

Article Snippet: We cultured the cells for 12 - 14 day in microglial differentiation medium which contains DMEM/F12, 2× insulin-transferrin-selenite, 2× B27, 0.5× N2, 1× glutamax, 1× non-essential amino acids (all from ThermoFisher Scientific), 400 μM monothioglycerol (Merck, Rahway, NJ, USA), 5 μg/mL human insulin (Merck), 100 ng/mL recombinant human IL-34 (Sino Biological, Beijing, China), 50 ng/mL human TGF-β1, and 25 ng/mL human M- CSF (all cytokines from Peprotech, Thermo Fisher Scientific).

Techniques: Staining, Co-Culture Assay, Infection