Methods and compositions for generating pacemaker cells

Abstract

Disclosed herein are methods and compositions for generating pacemaker cells from non-pacemaker cardiomyocytes. For example, the method includes the step of culturing the non-pacemaker cardiomyocytes with silk fibroin so that the silk fibroin induces the transformation of at least a portion thereof into pacemaker cells.

What is claimed is: 
     
         1 . Pacemaker cells, generated using a method comprising the step of,
 culturing a cell population comprising non-pacemaker cardiomyocytes with an effective amount of a silk fibroin biomaterial so that one or more of the non-pacemaker cardiomyocytes are transformed into the pacemaker cells.   
     
     
         2 . The pacemaker cells of  claim 1 , wherein the pacemaker cells comprise sinoatrial node (SAN)-like cells. 
     
     
         3 . The pacemaker cells of  claim 2 , wherein the SAN-like cells exhibit an increased expression level of hyperpolarization-activated, cyclic nucleotide-gated potassium channel 4 (HCN4) gene or connexin 45 gene, or both, compared with the non-pacemaker cardiomyocytes. 
     
     
         4 . The pacemaker cells of  claim 2 , wherein the SAN-like cells exhibit sinoatrial node-specific calcium clock. 
     
     
         5 . The pacemaker cells of  claim 1 , wherein the silk fibroin biomaterial is a silk fibroin solution, a silk fibroin particle, a non-woven silk fibroin mat, a silk fibroin hydrogel, a silk fibroin film, or a silk fibroin scaffold. 
     
     
         6 . The pacemaker cells of  claim 5 , wherein the silk fibroin film comprises,
 a biodegradable polymeric film having a plurality of amines group on the surface thereof; and   a plurality of silk fibroins crosslinked with the biodegradable polymeric film via the plurality of amine groups.   
     
     
         7 . A pharmaceutical composition for treating cardiac arrhythmia, comprising an effective number of pacemaker cells of  claim 1  and a pharmaceutically acceptable carrier thereof. 
     
     
         8 . A method for generating pacemaker cells, comprising the step of contacting a cell population comprising non-pacemaker cardiomyocytes with an effective amount of a silk fibroin biomaterial so that one or more of the non-pacemaker cardiomyocytes are transformed into the pacemaker cells. 
     
     
         9 . The method of  claim 8 , wherein the cell population and the silk fibroin biomaterial are contacted in vitro. 
     
     
         10 . The method of  claim 8 , wherein the cell population and the silk fibroin biomaterial are contacted in vivo. 
     
     
         11 . The method of  claim 8 , wherein the silk fibroin biomaterial is a silk fibroin solution, a silk fibroin particle, a non-woven silk fibroin mat, a silk fibroin hydrogel, a silk fibroin film, or a silk fibroin scaffold. 
     
     
         12 . The method of  claim 11 , wherein the silk fibroin film comprises,
 a biodegradable polymeric film having a plurality of amines group on the surface thereof; and   a plurality of silk fibroins crosslinked with the biodegradable polymeric film via the plurality of amine groups.   
     
     
         13 . A method for treating a subject suffering from cardiac arrhythmia, comprising the step of administering to the subject an effective number of pacemaker cells of  claim 1 . 
     
     
         14 . The method of  claim 13 , wherein the pacemaker cells are generated from non-pacemaker cardiomyocytes derived from the subject. 
     
     
         15 . The method of  claim 13 , wherein the silk fibroin biomaterial is a silk fibroin solution, a silk fibroin particle, a non-woven silk fibroin mat, a silk fibroin hydrogel, a silk fibroin film, or a silk fibroin scaffold. 
     
     
         16 . The method of  claim 13 , wherein the silk fibroin film comprises a biodegradable polymeric film having a plurality of amines group on the surface thereof, and a plurality of silk fibroins crosslinked with the biodegradable polymeric film via the plurality of amine groups. 
     
     
         17 . A method of treating a subject suffering from cardiac arrhythmia, comprising the step of administering to the heart of the subject an effective amount of a silk fibroin biomaterial. 
     
     
         18 . The method of  claim 17 , wherein the silk fibroin biomaterial is a silk fibroin solution, a silk fibroin particle, a non-woven silk fibroin mat, a silk fibroin hydrogel, a silk fibroin film, or a silk fibroin scaffold. 
     
     
         19 . The method of  claim 18 , wherein the silk fibroin film comprises a biodegradable polymeric film having a plurality of amines group on the surface thereof, and a plurality of silk fibroins crosslinked with the biodegradable polymeric film via the plurality of amine groups.