Ex vivo methods for validating substance testing with human organs and/or tissues
First Claim
Patent Images
1. An ex vivo method of validating results of substance testing using at least one human organ or tissue unsuitable for transplantation, wherein said human organ or tissue is a metabolically active human organ or tissue that has been permanently removed from its origin, the method comprising:
- (a) assessing said human organ or tissue for suitability for testing prior to administration of a test substance by perfusing said organ or tissue under physiological and normothermic conditions with a perfusate that comprises about 15 to about 20% (v/v) matched human erythrocytes, about 2 to about 6% human serum albumin, N-acetylcysteine and adenosine monophosphate (AMP), but does not comprise the test substance, wherein assessing said human organ or tissue for suitability includes determining that said human organ or tissue is unsuitable for transplantation, and eitherassaying the perfusate after it leaves said organ or tissue, ormonitoring a parameter of said organ or tissue;
(b) preparing a perfusate comprising said test substance, about 15 to about 20% (v/v) matched human erythrocytes, about 2 to about 6% human serum albumin, N-acetylcysteine and adenosine monophosphate (AMP), and perfusing said organ or tissue under said conditions with the perfusate containing the test substance;
(c) assessing a parameter relating to said test substance;
(d) assessing said organ or tissue for suitability for testing of said parameter relating to said test substance during and/or after administration of said test substance byquantifying a functional state of said organ or tissue relative to said parameter being assessed, wherein said organ or tissue acts as its own control,verifying that the organ or tissue is still having its effect, or being affected by, substances in connection with said parameter being assessed for the test substance, andverifying that said parameter being assessed is not inappropriately affected by an extraneous change in the organ or tissue during the course of assessment of the parameter relating to said test substance; and
wherein quantifying the functional state of said organ or tissue relative to said parameter being assessed in step (d) comprises perfusing said organ or tissue under same conditions with the same perfusate comprising about 15 to about 20% (v/v) matched human erythrocytes, about 2 to about 6% human serum albumin, N-acetylcysteine and adenosine monophosphate (AMP), and additionally containing at least one control substance having a known qualitative or quantitative effect with respect to said parameter, and assessing the same parameter with respect to at least one said control substance.
2 Assignments
0 Petitions
Accused Products
Abstract
Methods of validating results of assessments of test substances using human-derived tissues and/or organs, particularly tissues and/or organs unsuitable for transplantation, include assessing the suitability of the organ and/or tissue for substance testing, as well as inter-organ variability and use of exogenous and/or endogenous controls.
-
Citations
42 Claims
-
1. An ex vivo method of validating results of substance testing using at least one human organ or tissue unsuitable for transplantation, wherein said human organ or tissue is a metabolically active human organ or tissue that has been permanently removed from its origin, the method comprising:
-
(a) assessing said human organ or tissue for suitability for testing prior to administration of a test substance by perfusing said organ or tissue under physiological and normothermic conditions with a perfusate that comprises about 15 to about 20% (v/v) matched human erythrocytes, about 2 to about 6% human serum albumin, N-acetylcysteine and adenosine monophosphate (AMP), but does not comprise the test substance, wherein assessing said human organ or tissue for suitability includes determining that said human organ or tissue is unsuitable for transplantation, and either assaying the perfusate after it leaves said organ or tissue, or monitoring a parameter of said organ or tissue; (b) preparing a perfusate comprising said test substance, about 15 to about 20% (v/v) matched human erythrocytes, about 2 to about 6% human serum albumin, N-acetylcysteine and adenosine monophosphate (AMP), and perfusing said organ or tissue under said conditions with the perfusate containing the test substance; (c) assessing a parameter relating to said test substance; (d) assessing said organ or tissue for suitability for testing of said parameter relating to said test substance during and/or after administration of said test substance by quantifying a functional state of said organ or tissue relative to said parameter being assessed, wherein said organ or tissue acts as its own control, verifying that the organ or tissue is still having its effect, or being affected by, substances in connection with said parameter being assessed for the test substance, and verifying that said parameter being assessed is not inappropriately affected by an extraneous change in the organ or tissue during the course of assessment of the parameter relating to said test substance; and wherein quantifying the functional state of said organ or tissue relative to said parameter being assessed in step (d) comprises perfusing said organ or tissue under same conditions with the same perfusate comprising about 15 to about 20% (v/v) matched human erythrocytes, about 2 to about 6% human serum albumin, N-acetylcysteine and adenosine monophosphate (AMP), and additionally containing at least one control substance having a known qualitative or quantitative effect with respect to said parameter, and assessing the same parameter with respect to at least one said control substance. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42)
-
Specification