Data analysis of cells undergoing excitation contraction coupling as detected on real-time cell analysis (rtca) instruments

Abstract

A method of determining a beating parameter of cells that undergo excitation contraction coupling, the method including providing a cell analysis device having a substrate and a sensor that measures cell adhesion or attachment to the substrate in millisecond time resolution; adding excitable cells capable of undergoing excitation contraction coupling to the substrate; monitoring cell adhesion or attachment of the excitable cells to the substrate in millisecond time resolution; and calculating one or more beating parameters from the monitored adhesion.

What is claimed is: 
     
         1 . A method of determining a beating parameter of cells that undergo excitation contraction coupling, the method comprising:
 a) providing a cell analysis device comprising a substrate and a sensor that measures cell adhesion or attachment to the substrate in millisecond time resolution;   b) adding excitable cells capable of undergoing excitation contraction coupling to the substrate;   c) monitoring cell adhesion or attachment of the excitable cells to the substrate in millisecond time resolution; and   d) calculating one or more beating parameters from the monitored adhesion.   
     
     
         2 . The method according to  claim 1 , wherein the cell analysis device is an impedance monitoring device that measures cell-substrate impedance. 
     
     
         3 . The method according to  claim 1 , wherein the cells are cardiomyocytes. 
     
     
         4 . The method according to  claim 1 , wherein the cells are stem cell derived cardiomyocytes. 
     
     
         5 . The method according to  claim 1 , wherein the cells are induced pluripotent stem cell (iPS cell) derived cardiomyocytes. 
     
     
         6 . The method according to  claim 1 , wherein the millisecond resolution is characterized as consecutive impedance measurements less than 40 milliseconds apart. 
     
     
         7 . The method according to  claim 6 , wherein the millisecond resolution is characterized as consecutive impedance measurements less than 20 milliseconds apart. 
     
     
         8 . The method according to  claim 1 , wherein the step of calculating one or more parameters comprises forming a beating curve from the monitored cell adhesion or attachment and deriving the one or more beating parameters from the beating curve. 
     
     
         9 . The method according to  claim 1 , wherein the one or more beating parameters are selected from the group consisting of beating rate, beating amplitude, rising time, falling time, beating period, IBD10, IBD50, IBD90, rising slope and falling slope. 
     
     
         10 . The method according to  claim 1 , wherein the one or more beating parameters are selected from the group consisting of normalized beating rate, normalized beating amplitude, beating pattern similarity and beating rhythm irregularity. 
     
     
         11 . The method according to  claim 1 , wherein the one or more beating parameters comprises 5 beating parameters. 
     
     
         12 . The method according to  claim 1 , wherein the one or more beating parameters are calculated while the device continues to monitor the cell adhesion or attachment. 
     
     
         13 . The method according to  claim 1 , further comprising adding a test compound to the cells, wherein the step of monitoring cell adhesion or attachment of the excitable cells is performed after adding the test compound. 
     
     
         14 . The method according to  claim 13 , wherein the step of monitoring cell adhesion or attachment of the excitable cells is performed before and after adding the test compound. 
     
     
         15 . A method of determining a dose response for a test compound on a cell population, the method comprising:
 a) determining a beating parameter for different doses of a test compound according to the method of  claim 1 , wherein the test compound is added at different doses to different populations of cells; and   b) plotting the beating parameters for each dose to form a dose response curve.   
     
     
         16 . The method according to  claim 15 , wherein the beating parameter for each dose is from a same time point. 
     
     
         17 . The method according to  claim 15 , further comprising determining a IC50 or EC50 value from the dose response curve. 
     
     
         18 . A method for identifying a compound having a potentially cardiotoxic effect, comprising:
 a) providing a test compound suspected of having a cardiotoxic effect;   b) performing the method according to  claim 1  to obtain a beating parameter for the test compound, wherein the cells are cardiomyocytes and wherein the test compound is added to the cardiomyocytes; and   c) comparing the beating parameter for the test compound to a control beating parameter to identify whether there is a difference between beating parameters and if so, concluding the test compound has the potentially cardiotoxic effect.   
     
     
         19 . The method according to  claim 18 , wherein the beating parameter for the test compound comprises 2 beating parameters, which are compared to 2 control beating parameters, further wherein the test compound is concluded to have a cardiotoxic effect if at least one of the compared parameters is different. 
     
     
         20 . The method according to  claim 18 , wherein the compound is suspected of being a pro-arrhythmic drug that may induce arrhythmia. 
     
     
         21 . The method according to  claim 18 , wherein the step of monitoring the beating of cells is also performed before adding the test compound. 
     
     
         22 . The method according to  claim 18 , wherein the test compound is added at different doses to the cells.