Extended Data Fig. 8: Optimization of tri-culture ratio and culture media to maximize cell survival and minimize cell activation at baseline. | Nature Neuroscience

Extended Data Fig. 8: Optimization of tri-culture ratio and culture media to maximize cell survival and minimize cell activation at baseline.

From: Fully defined human pluripotent stem cell-derived microglia and tri-culture system model C3 production in Alzheimer’s disease

Extended Data Fig. 8

a, IF shows that tri-cultures with increased numbers of astrocytes plated (50 K) show fewer microglial cells (IBA1 + ) attached at Day 7 than with tri-cultures containing fewer (25 K) astrocytes. Scale bar = 200 µM. b, NB/BAGC and NB:N2 base media formulations show the lowest secretion of C3 in the tri-culture by ELISA. Addition of DMEM base, F12 supplement, low glucose, high glutamine, and high pyruvate all increase C3 levels in baseline tri-cultures. n = 2 technical replicates of cell culture supernatant. c, Control staining for CC3. IF of hPSC-derived neurons killed by 70% methanol incubation for 30 minutes shows bright CC3 + whereas IF of fixed cells without the methanol treatment does not. Scale bar = 100 µM.

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