Background: Important species differences make a human model of in vitro hypoxic neuronal cell death desirable.

Materials and Methods: Human NT2 teratocarcinoma cells were differentiated to NT2-N neurons and exposed to hypoxia for 6 hours. Cell death was evaluated after 48 hours by microscopy and by LDH release (% of total LDH).

Results: Marked cell death was seen in hypoxic cultures. LDH release after hypoxia was 24.2±5.5% vs 13.8±3.7% in nonhypoxic controls (p<0.01). If glutamine was omitted from the medium (preventing glutamate release) or the NMDA receptor blocker MK-801 was added before hypoxia, cell death was almost completely prevented (LDH release 15.4±4.5% and 15.5±5.2%, p<0.001). The non-NMDA receptor blocker CNQX also provided marked protection (18.7±5.1%, p<0.001) while the 21-aminosteroid U74389G, N-acetylcystein, the spin trap PBN or the nitric oxide synthase inhibitor L-nitroarginine were ineffective.

Conclusion: Both NMDA and non-NMDA receptors are important for hypoxic cell death in human NT2-N neurons.