Establishment of neuronal in vitro models of ischemia in 96-well microtiter strip-plates that result in acute, progressive and delayed neuronal death

HAMI 3379, a CysLT2 Receptor Antagonist, Attenuates Ischemia-Like Neuronal Injury by Inhibiting Microglial Activation

“…Primary cortical neurons were isolated from neonatal Sprague-Dawley rats of either sex within 24 hours of birth (Laboratory Animal Center of Zhejiang University, Hangzhou, China) as described previously with modifications (Meloni et al, 2001). In brief, after quick decollation using mechanical shearing, the cerebral cortices were dissected and digested with 0.25% trypsin (Sangon, Shanghai, China) for 10 minutes at 37°C.…”

A multifunctional cytoprotective agent that reduces neurodegeneration after ischemia

“…H͞H was produced in vitro by reducing the concentration of glucose to 1.2 mM by diluting the culture medium with degassed balanced salt solution (BSS) (28) and O 2 to 0% in a sealed container filled with 5% CO 2 and 95% N 2 . Control specimens were treated with nondegassed BSS containing 25 mM glucose and ambient air with 5% CO 2 .…”

“…Control specimens were treated with nondegassed BSS containing 25 mM glucose and ambient air with 5% CO 2 . After 6 h, the solutions in both hypoxic and normoxic cultures were diluted with a termination solution (28) and further cultured in 5% CO 2 and 95% ambient air for another 24 or 48 h. In the hypoxia-only study, the plates were incubated in 95% N 2 and 5% CO 2 at 37°C for 18 h in normal culture medium. Control (normoxic) plates were incubated in ambient air with 5% CO 2 for the same time period.…”

Neuronal injury in NCX3 knockout mice following permanent focal cerebral ischemia and in NCX3 knockout cortical neuronal cultures following oxygen-glucose deprivation and glutamate exposure.

“…The longer and milder oxygen glucose deprivation insult used by Molinaro et al (2008) is likely to be related to the use of plastic culture vessels which are known to store oxygen (Meloni et al, 2001) and thus supplying oxygen to neurons for energy generation and maintenance of ionic gradients.…”

“…Primary cortical neuronal cultures were prepared from NCX3 knockout and wildtype embryos using methods as previously described for rat embryos (Meloni et al, 2001). Briefly, cortical brain tissue from E17 embryos was dissociated in Dulbelcco's Modified Eagle Medium (DMEM; Invitrogen, Australia) supplemented with 1.3 mM L-cysteine, 0.9 mM NaHCO 3 , 10 units/ml papain (Sigma) and 50 units/ml DNase (Sigma) and washed in cold DMEM/10% horse serum.…”