Multiloop Engineered Heart Muscle Tissue

Abstract

The invention is directed to a method for the preparation of a multiring engineered heart tissue construct suitable for use in cardiac tissue augmentation and/or replacement therapy. The invention further refers to multiring EHT constructs which comprise at least two force-generating engineered heart tissue rings fused with each other and a device for preparing the same. Finally, the invention relates to force-generating engineered heart tissue rings derived from human cells and their use in drug screening and target validation assays.

1 . Method for the ex vivo-preparation of a multiring engineered heart tissue construct suitable for use in cardiac tissue augmentation and/or replacement therapy, comprising the steps of
 a) providing force-generating engineered heart tissue rings;   b) placing in position at least two force-generating engineered heart tissue rings so that each force-generating engineered heart tissue ring has one or more contact points to an adjacent force-generating engineered heart tissue ring; and   c) culturing the force-generating engineered heart tissue rings under conditions which allow fusion of the at least two force-generating engineered heart tissue rings at the one or more contact points to form a multiring heart muscle construct.   
   
   
       2 . Method of  claim 1 , wherein the method further comprises subjecting the at least two force-generating engineered heart tissue rings to tensile stress prior to or simultaneously with step b). 
   
   
       3 . Method of  claim 2 , wherein the at least two force-generating engineered heart tissue rings are subjected to tensile stress in a static, phasic or auxotonic manner or a combination thereof. 
   
   
       4 . Method of  claim 3 , wherein the at least two force-generating engineered heart tissue rings are subjected to tensile stress in an auxotonic manner. 
   
   
       5 . Method of  claim 4 , wherein the tensile stress is applied by elastically suspending each of the at least two force-generating engineered heart tissue rings between at least two associated suspension means, wherein at least one of said associated suspension means is resiliently biased so that the individual force-generating engineered heart tissue rings are able to contract against the bias force provided by the suspension means and are reexpanded by this force during relaxation. 
   
   
       6 . Method of  claim 5 , wherein each of the at least two force-generating engineered heart tissue rings are suspended between two associated suspension means. 
   
   
       7 . Method of  claim 5  or  6 , wherein all associated suspension means are resiliently biased. 
   
   
       8 . Method of  claims 5  to  7 , wherein the at least one suspension means can be adjusted to vary the tensile stress. 
   
   
       9 . Method of  claims 4  or  8 , wherein tensile stress is applied simultaneously with step b). 
   
   
       10 . Method of  claims 2  to  9 , wherein culturing in step c) is conducted while the at least two force-generating engineered heart tissue rings are under tensile stress. 
   
   
       11 . Method of any of the preceding claims, wherein 5 or more force-generating engineered heart tissue rings are placed into contact. 
   
   
       12 . Method of any of the preceding claims, wherein the force-generating engineered heart tissue rings have an outer diameter of 8-12 mm. 
   
   
       13 . Method of any of the preceding claims, wherein the force-generating engineered heart tissue rings are placed into contact by stacking to form a central region in which the force-generating engineered heart tissue rings overlap with each other. 
   
   
       14 . Method of any of the preceding claims, wherein the force-generating engineered heart tissue rings are derived from human cells. 
   
   
       15 . Method of  claim 14 , wherein the engineered heart tissue rings comprises cells comprising one or more of the following proteins: cardiac myosin heavy chain, α-actinin, desmin and/or cardiac troponin I. 
   
   
       16 . Multiring engineered heart tissue construct comprising at least two force-generating engineered heart tissue rings fused with each other. 
   
   
       17 . Multiring engineered heart tissue construct of  claim 16 , comprising 5 or more force-generating engineered heart tissue rings fused with each other to form an interconnected tissue construct. 
   
   
       18 . Multiring engineered heart tissue construct of  claim 16  or  17 , wherein the force-generating engineered heart tissue rings have an outer diameter of 8-12 mm. 
   
   
       19 . Multiring engineered heart tissue construct of  claims 16  to  18 , wherein the force-generating engineered heart tissue rings are stacked to form a central region in which the force-generating engineered heart tissue rings overlap with each other. 
   
   
       20 . Multiring engineered heart tissue construct of  claims 16  to  19 , wherein the force-generating engineered heart tissue rings are derived from human cells. 
   
   
       21 . Multiring engineered heart tissue construct of any of the preceding claims having a twitch tension of more than 2.5 mN. 
   
   
       22 . Multiring engineered heart tissue construct of  claim 21 , having a twitch tension of more than 3 mN. 
   
   
       23 . Multiring engineered heart tissue construct of any of the preceding claims wherein the construct comprises cells comprising one or more of the following proteins: cardiac myosin heavy chain, α-actinin, desmin and/or cardiac troponin I. 
   
   
       24 . Multiring engineered heart tissue construct, obtainable according to a method of  claims 1  to  15 . 
   
   
       25 . Device for preparing a multiring engineered heart tissue construct, comprising a plurality of associated suspension means for suspending force-generating engineered heart tissue rings under tensile stress, wherein at least one of each of the associated suspension means is resiliently biased so that the suspended force-generating engineered heart tissue ring is able to contract against the bias force provided by the suspension means, wherein the suspension means are arranged to each other so that each force-generating engineered heart tissue ring has one or more contact points to an adjacent force-generating engineered heart tissue ring when suspended in the device. 
   
   
       26 . Device of  claim 25 , wherein two suspension means are associated to each other. 
   
   
       27 . Device of  claim 25  or  26 , wherein all associated suspension means are resiliently biased. 
   
   
       28 . Device of  claim 25  to  27 , wherein the at least one suspension means can be adjusted to vary the tensile stress. 
   
   
       29 . Device of  claim 25  to  27 , wherein the associated suspension means are arranged to each other so that the force-generating engineered heart tissue rings can be suspended by stacking to form a central region in which the force-generating engineered heart tissue rings overlap with each other. 
   
   
       30 . Force-generating engineered heart tissue ring derived from human cells. 
   
   
       31 . Use of the force-generating engineered heart tissue ring of  claim 30  in drug screening or target validation assays.