In vitro ischemia model
View Patent ↗A tissue culture model of oxygen/glucose deprivation induced cell death is provided, which is useful in the analysis of the mechanisms of cell death following brain ischemia, and for screening anti-ischemic drugs. By adopting the in vivo concentrations of calcium, potassium and hydrogen ions to the incubation medium a model is established that shows conspicuous similarities with the temporal and special development of cell death in vivo: selective and delayed CA1 damage, a damage mitigated by blockade of the NMDA and AMPA receptors, and a striking augmentation of damage by high levels of glucose.
1. An ischemic cell culture, comprising:
a culture comprising hippocampal tissue, and a medium for simulation of ischemia, comprising potassium at a concentration of from about 50 mM to 90 mM; calcium at a concentration of from about 0.1 mM to 1 mM; NaCl at a concentration of from about 30-77 mM; at a pH of from 6.7 to 6.9,
wherein when said cells are exposed to said medium for a period of time sufficient to mimic the effects of ischemia in vivo the ischemic effects include a selective and delayed, glutamate receptor dependent cell death in the CA1 region.
2. The method according to claim 1 , wherein said cells comprise a section of brain tissue.
3. An ischemic cell culture, comprising:
a culture comprising hippocampal tissue, and a glucose free medium for simulation of ischemia, comprising potassium at a concentration of from about 50 mM to 90 mM; calcium at a concentration of from about 0.1 mM to 1 mM; at a pH of from 6.7 to 6.9,
wherein when said cells are exposed to said medium for a period of time sufficient to mimic the effects of ischemia in vivo the ischemic effects include a selective and delayed, glutamate receptor dependent cell death in the CA1 region.
4. The culture according to claim 1 , wherein glucose is present in said medium.
5. A method of simulating ischemia in vitro, the method comprising:
exposing a culture comprising hippocampal tissue to a medium for simulation of ischemia, said medium comprising potassium at a concentration of from about 50 mM to 90 mM; calcium at a concentration of from about 0.1 mM to 1 mM; at a pH from 6.7 to 6.9,
wherein when said cells are exposed to said medium for a period of time sufficient to mimic the effects of ischemia in vivo the ischemic effects include a selective and delayed, glutamate receptor dependent cell death in the CA1 region.
6. The method according to claim 1 , wherein said cells comprise a section of brain tissue.
7. The method according to claim 1 , wherein said medium is glucose free.
8. The method according to claim 1 , wherein glucose is present in said medium.
9. A method for determining the effect of a candidate agent on ischemia, the method comprising:
exposing a culture comprising hippocampal tissue to said candidate agent before, during, or after said cells are exposed to a medium for simulation of ischemia, said medium comprising potassium at a concentration of from about 50 mM to 90 mM; calcium at a concentration of from about 0.1 mM to 1 mM; at a pH of from 6.7 to 6.9 ,
wherein when said cells are exposed to said medium for a period of time sufficient to mimic the effects of ischemia in vivo the ischemic effects include a selective and delayed, glutamate receptor dependent cell death in the CA1 region;
comparing at least one parameter of said cells in the presence of said candidate agent and in a control wherein said agent is absent.
10. The method according to claim 9 , wherein said cells comprise a section of brain tissue.
11. The method according to claim 9 , wherein said medium is glucose free.
12. The method according to claim 9 , wherein glucose is present in said medium.