Effect of Trophic Factors on Delayed Neuronal Death Induced by in Vitro Ischemia in Cultivated Hippocampal and Cortical Neurons

Abstract

The effect of trophic factors on neuronal survival after 30 min oxygen and glucose deprivation (in vitro ischemia) was studied in primary hippocampal and cortical neuronal cultures of rat. In vitro ischemia was produced at 37 degrees C by placing cultures in glucose-free medium, the oxygen content of which was removed by gassing with pure argon. After in vitro ischemia neurons were allowed to recover either in serum-free minimal essential medium (MEM) or in MEM containing 5% native horse serum, 100 ng/ml basic fibroblast growth factor (bFGF) or 10 ng/ml transforming growth factor-beta 1 (TGF-beta 1), respectively. Cultures that recovered in serum-free medium suffered a progressive type of neuronal injury: survival of either cortical or hippocampal neurons declined from about 60% after 1 h to 50% after 3 h, 40% after 6 h and less than 20% after 24 h. Addition of serum proteins to the incubation medium did not influence early survival (up to 3-6 h) but significantly improved survival after 24 h (more than 40% in both hippocampal and cortical cultures). Addition of TGF-beta 1 and bFGF had only minor effects. These data show that serum reduces delayed ischemic cell death by a mechanism which is different from that of TGF-beta 1 or bFGF protection.

Rapid Ischemic Cell Death in Immature Oligodendrocytes: A Fatal Glutamate Release Feedback Loop

“…The [Ca 2ϩ ] i increase was apparent within 1 min of the onset of ischemia in most cells and continued to increase for 5-10 min before reaching a plateau. Cell death became evident within 5 min of the onset of ischemia, and the mean time to cell death was 20.4 min with 80% of the cells dead after 31 min; this represents the highest sensitivity to ischemic injury in any CNS cell type yet studied (Petito and Pulsinelli, 1984;Goldberg and Choi, 1993;Johnson et al, 1994;Pantoni et al, 1996;Kusumoto et al, 1997;Lyons and Kettenmann, 1998;Petito et al, 1998).…”

Primary culture of cellular subtypes from postnatal mouse for in vitro studies of oxygen glucose deprivation

“…Durations of OGD as brief as 10-30 min have been found to injure (Goldberg and Choi, 1993;Dugan et al, 1995), or kill cells in neuronal-enriched cultures (Goldberg and Choi, 1993;Dugan et al, 1995;Kusumoto et al, 1996Kusumoto et al, , 1997. However, other studies have reported that oxygen glucose deprivation of 60-240 min or more is required to produce substantial cell death in neuronal cultures (Vibulsreth et al, 1987).…”

Pyruvate protects glucose‐deprived Müller cells from nitric oxide‐induced oxidative stress by radical scavenging

“…conditions were achieved by flushing with a gas mixture of 95% argon/5% CO 2 (v/v) (Kusomoto et al, 1997). A stock solution of 100 mM papanonoate in 20 mM NaOH was prepared and added to the medium in a final concentration of 1 mM (Thomas et al, 2002).…”