In vitro ischemia model
AssigneeAGY Therapeutics, Inc.
InventorsTadeusz Wieloch, Anna Rytter, Tobias Cronberg
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Abstract
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.
Claims32
A method of simulating ischemia in vitro, the method comprising:
- exposing cultured cells to 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;
- at a pH of from 6.7 to 6.9, wherein said cells mimic the effects of ischemia in vivo.
A method for determining the effect of a candidate agent on ischemia, the method comprising:
- exposing cultured cells said candidate agent before, during or after said cells are exposed to 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;
- at a pH of from 6.7 to 6.9, wherein said cells mimic the effects of ischemia in vivo;
- comparing at least one parameter of said cells in the presence of said candidate agent and in a control wherein said agent is absent.
A method for detecting genetic sequences that are differentially expressed in ischemia, the method comprising:
- exposing cultured cells to 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;
- at a pH of from 6.7 to 6.9, wherein said cells mimic the effects of ischemia in vivo;
- isolating expressed mRNA from said cells;
- and
- comparing said mRNA to mRNA obtained from a control cell culture.
A glucose free sterile medium for simulation of ischemia, consisting essentially of:
- 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.
An ischemic cell culture, comprising:
- cultured cells, 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;
- at a pH of from 6.7 to 6.9, wherein said cells mimic the effects of ischemia in vivo.
Dependencies are inferred from the claim text.