Isolated intestinal mucosa and uses thereof
AssigneeAlbert LI
InventorAlbert LI
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Abstract
The present disclosure provides an in vitro reagent for evaluating xenobiotic metabolism in a cell culture based assay. The in vitro reagent is an admixture of a cell culture medium and isolated intestinal mucosa comprising villi wherein the intestinal mucosa was eluted from a lumen of the intestine. The isolated mucosa comprises metabolically competent cells. Addition of a xenobiotic test compound to the in vitro reagent allows metabolism of the test compound by the isolated intestinal mucosa comprising villi.
Claims25
A method of manufacturing metabolically competent isolated intestinal mucosal comprising villi eluted from intestines, comprising:
- a) eluting the intestinal mucosal from lumen of the intestines to form isolated intestinal mucosa;
- b) suspending the isolated intestinal mucosa in a cell culture medium;
- c) adding a cryoprotectant to the cell culture medium to form an in vitro reagent;
- and,
- d) storing the in vitro reagent frozen at a temperature of −10° C. to about −175° C.
An in vitro reagent for evaluating biological activity of a test substance, wherein the reagent is a cryopreserved mixture comprising:
- a) a cell culture medium comprising a cryoprotectant;
- and,
- b) metabolically competent isolated intestinal mucosa comprising villi wherein the intestinal mucosa was eluted from an intestine.
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17-33. (canceled)
A method for evaluating in vitro inhibition or induction of cytochrome P450 by a test compound that is ingested or administered orally, comprising:
- a) providing metabolically competent isolated intestinal mucosa comprising villi wherein the intestinal mucosa was eluted from an intestine and previously cryopreserved;
- b) culturing the isolated intestinal mucosa in a cell culture vessel incubated at 30-45° C.;
- c) introducing a test compound into the cell culture vessel that is suspected of being a P450 ligand;
- d) incubating the test compound for 1 minute to 10 days at 30-45° C.;
- and,
- e) performing an end point assay of the isolated intestinal mucosa or cell culture medium to determine P450 activity.
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41-50. (canceled)
Dependencies are inferred from the claim text.