Fig. 4

Different cell culture substrates were assessed for optimizing NCM culture. a Morphology of NCM114 (late passage, P4) seeded on a 24-well plate coated without or with 80 μL of Matrigel™ per well at 18,333 cells per well. Scale bars = 100 μm. b Morphology of NCMs on different substrates. In a 96-well plate, 2500 cells were seeded per well and were imaged after 3 days. Scale bars = 100 μm. c The metabolic activity of NCMs was assessed after 3 days of culture on different substrates. Data are shown as mean ± SD. One-way ANOVA with multiple comparisons test (baseline: no coating), ns: not significant. **P < 0.01, ****P < 0.0001. d Morphology of NCMs on Matrigel™ at various cell seeding densities. In a 96-well plate coated with 45 μL of Matrigel™ per well, 1000, 3000, 5000, or 7000 cells were seeded per well and were imaged after 7 days. b–d n = 4 biological replicates (NCMs from 4 different donors), with each repeated in n = 3 wells. Scale bars = 400 μm