CO-CULTURED SKELETAL MYOCYTE ANDCARDIOMYOCYTE CELL-SHEETS COULD NOT ESTABLISHELECTRICAL COMMUNICATION, BUT CAUSEDFIBRILLATING ACTIVITY IN CARDIOMYOCYTE

摘要

Transplantation to the patient with enzymatically isolated skeletal myocyte (SM) cell improved cardiac function and caused serious arrhythmias, despite electrical disconnection between the SM and host cardiomyocyte (CM) was shown. Previously we reported cell culture surfaces grafted with temperature-responsive polymer, from which confluent cells detach as a cell-sheet simply by reducing temperature without any enzymatic treatments. We applied the cell-sheet technology for the transplantation to establish electrical communication between SM and CM and observed arrhythmogeneity. Methods Prepared neonate rat derived SM and CM cell-sheets were co-cultured for 3 days. The action potential was observed by optical image of voltage sensitive dye to test electrical communication between the SM-sheet and CM-sheet. The spontaneous and arrhythmic contractile activity were monitored by video motion-detecting program and [Ca~(2+)]i imaging. Results No electrical communication between SM and CM-sheet (0/20) was observed. Spontaneous and rhythmic contraction was observed in SM (24/30). Fibrillating contraction was observed in the CM-sheet, when it was co-cultured with SM-sheet. Conclusions Our cell-sheet graft technology failed to establish the electrical connection. Fibrillating contraction in CM was observed for the first time when we co-cultured with SM-sheet. Automaticity in SM might be foe for the arrhythmogenesis via activating stretch activated channel in CM.