Patterned cardiomyocyte culture on microelectrode array

Abstract

The invention provides a culture of patterned cardiomyocytes, the culture including a support substrate bearing a multielectrode array, a negative surface resistant to cell attachment and deposited on the support substrate covering the multi-electrode array, a pattern ablated on the negative surface, a positive surface promoting cell attachment deposited on the pattern ablated on the negative surface; and cardiomyocytes adherent to the positive surface and growing aligned along the pattern ablated on the negative surface. The invention also includes a method of making the culture of patterned cardiomyocytes and a method of in vitro method of testing the effect of a compound on cardiac function by measuring electrical output from the patterned cardiomyocytes.

1 - 10 . (canceled) 
     
     
         11 . A culture of patterned cardiomyocytes, said culture comprising:
 i) a support substrate bearing a multielectrode array;   ii) a first surface resistant to cell attachment and deposited on said support substrate wherein the first surface covers the multi-electrode array;   iii) a pattern ablated on said first surface;   iv) a second surface that promotes cell attachment deposited on the pattern ablated on the first surface; and   v) cardiomyocytes adherent to said second surface and growing aligned along said pattern to provide said culture of patterned spontaneously beating cardiomyocytes.   
     
     
         12 . The culture of  claim 11 , wherein the first surface comprises at least one material selected from the group consisting of polyethylene glycol, polyacrylic acid and polyacrylamide. 
     
     
         13 . The culture of  claim 12 , wherein the material selected is polyethylene glycol deposited essentially as a monolayer. 
     
     
         14 . The culture of claim  10 , wherein the second surface comprises at least one material selected from the group consisting of fibronectin, trimethoxysilylpropyldiethylenetriamine and pure glass. 
     
     
         15 . The culture of claim  10 , wherein the pattern is ablated by photolithography. 
     
     
         16 . The culture of claim  10 , wherein the ablated pattern comprises a path extending over two or more electrodes of the multielectrode array. 
     
     
         17 . The culture of  claim 16 , wherein the support substrate comprises both an multielectrode array-bearing portion and a portion devoid of a multielectrode array, the first surface is deposited on both the multielectrode array-bearing portion and the portion devoid of a multielectrode array, and the path extends over both the multielectrode array-bearing portion and the portion devoid of a multielectrode array. 
     
     
         18 . The culture of  claim 16 , wherein the path comprises a first end, a middle region, and a second end, wherein the second end comprises a branched region. 
     
     
         19 . The culture of  claim 18 , wherein the first end of the path contacts at least two electrodes of the multielectrode array, the middle region of the path extends over the portion of the support substrate devoid of the multielectrode array, and the branched region extends over at least two electrodes of the multielectrode array. 
     
     
         20 . The culture of  claim 19 , wherein the branched region comprises a first branch and a second branch in a Y-shaped configuration, and each of the first branch and the second branch extend over at least two electrodes of the multielectrode array. 
     
     
         21 . The culture of claim  10 , wherein the cardiomyocytes are of mammalian origin. 
     
     
         22 . The culture of claim  10 , wherein the cardiomyocytes are of rat origin. 
     
     
         23 . The culture of claim  10 , wherein the cardiomyocytes are derived from human embryonic stem cells.