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24 ((triculture media, hiNSC, Astrocyte Growth medium, MEF Feeder Media:…
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hiNSC
KnockOut DMEM w 20% CTS KnockOut Sr XenoFree Medium, 1% Glutamax, 1% anti-anti, 0.1% 2-mercaptoethanol, 20ng/mL human bFGF
Astrocyte Growth medium
2% FBS, 1% P/S, 1% astrocyte growth supplement
MEF Feeder Media: DMEM w 10% FBS, 1% antibiotic-antimycotic
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Microglia (HMC3) medium: Eagle's Minimum Essential Medium supplemented w 10% heat-inactivaed FBS and 1% anti-anti
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Triculture
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seeding volume: cells were seeded in a total volume of 40uL of triculture media per scaffold, addition of 150uL of media after initial attachment
maturation timeline: cultured for 6 weks prior to injury testing to allow hiNSCs to mature into neurons
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