Various methods have been developed to differentiate cardiomyocytes from human pluripotent stem cells. In addition, the studies related to the maturation of differentiated cardiomyocytes have been conducted. However, despite these efforts, the differentiated cardiomyocytes are much less mature than ...
Various methods have been developed to differentiate cardiomyocytes from human pluripotent stem cells. In addition, the studies related to the maturation of differentiated cardiomyocytes have been conducted. However, despite these efforts, the differentiated cardiomyocytes are much less mature than the actual adult heart. Previous studies of cardiomyocytes differentiation using Matrigel-based protocols did not provide a cardiac-specific environment for cardiomyocytes, since Matrigel is composed of the solubilized basement membrane matrix secreted by mouse cancer cells. Therefore, it is necessary to develop new cardiac differentiation protocol with cardiac tissue-derived extracellular matrix. Here, we demonstrate new cardiac differentiation method using porcine heart-derived decellularized extracellular matrix (dECM) and a cardiac patch. The heart-derived dECM can provide a cardiac-specific environment for cardiomyocytes, and, the cardiac patch method induces structural and functional maturation of cardiomyocytes, enabling dynamic beating not seen in two dimensional culture. Human embryonic stem cell-derived cardiomyocytes were embedded in the patches which were composed of dECM or Matrigel, and the cardiomyocytes in the cardiac patches were viable during fabrication of cardiac patch and subsequent cultivation. The expression of cardiomyocyte-specific markers, including α-SA and cTnT, in the dECM-derived cardiac patch was greater than the Matrigel-derived cardiac patch. Furthermore, the expression of cTnI, a mature cardiomyocyte marker, was also increased in the dECM-derived cardiac patch. Moreover, the expression of a calcium ion channel, CACNA1A, and a cardiac sarcoplasmic reticulum Ca2+-ATPase, SERCA2a, was also increased in the dECM-derived patch than the Matrigel-derived patch. In conclusion, cardiac patch fabrication using dECM which provides a heart-specific environment for cardiomyocytes can induce the maturation of cardiomyocytes, and it can be applied drug and toxicity assessments.
Various methods have been developed to differentiate cardiomyocytes from human pluripotent stem cells. In addition, the studies related to the maturation of differentiated cardiomyocytes have been conducted. However, despite these efforts, the differentiated cardiomyocytes are much less mature than the actual adult heart. Previous studies of cardiomyocytes differentiation using Matrigel-based protocols did not provide a cardiac-specific environment for cardiomyocytes, since Matrigel is composed of the solubilized basement membrane matrix secreted by mouse cancer cells. Therefore, it is necessary to develop new cardiac differentiation protocol with cardiac tissue-derived extracellular matrix. Here, we demonstrate new cardiac differentiation method using porcine heart-derived decellularized extracellular matrix (dECM) and a cardiac patch. The heart-derived dECM can provide a cardiac-specific environment for cardiomyocytes, and, the cardiac patch method induces structural and functional maturation of cardiomyocytes, enabling dynamic beating not seen in two dimensional culture. Human embryonic stem cell-derived cardiomyocytes were embedded in the patches which were composed of dECM or Matrigel, and the cardiomyocytes in the cardiac patches were viable during fabrication of cardiac patch and subsequent cultivation. The expression of cardiomyocyte-specific markers, including α-SA and cTnT, in the dECM-derived cardiac patch was greater than the Matrigel-derived cardiac patch. Furthermore, the expression of cTnI, a mature cardiomyocyte marker, was also increased in the dECM-derived cardiac patch. Moreover, the expression of a calcium ion channel, CACNA1A, and a cardiac sarcoplasmic reticulum Ca2+-ATPase, SERCA2a, was also increased in the dECM-derived patch than the Matrigel-derived patch. In conclusion, cardiac patch fabrication using dECM which provides a heart-specific environment for cardiomyocytes can induce the maturation of cardiomyocytes, and it can be applied drug and toxicity assessments.
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