| that of 3.Result:1.Most of cardiac myocytes isolated by trypsin and collagenaseⅡwere alive.The cells contracted rhythmically after 48h in vitro,meanwhile,they expressedα-sacromeric actin and desmin positively.Cardiomyocytes got together and beat synchronously in clusters gradually after 3 d.The vitality of cardiac myocytes kept stable from 3 to 15 days in culture with the density of 2×105/cm2 and 4×105/cm2.2.The loading device with fundus run successfuly for 1 week.Temperature and programme displayed normally.When the strain of silicone membrane was 0～30%,the reproducibility of load-displacement curve was good.Cells cultured in the device were alive and no bacterial contamination in 2 weeks.3.After cyclic stretch,cell morphous changed from polygon to rod shape,and the orientating of cells altered.After 4d,the angle of cell macroaxis and stretch direction trended to 45°or 90°when strain at 8%,1 Hz.LDH leaking and PI staining positive ratio of(1Hz,12%)strain were more than that of others.When the strain was 4%,the QGluand QLacwere more than that of static group,and changes were significance when the strain was 8%(P＜0.05).If cells were overloaded,the QGluand QLacwere less than that of static group.The result of Western Blot showed that microtubule mass increased more in 8%strain than other strain levels.When the strain was 12%,level of microtubule mass lower than that of 8%.Fixed the strain at 8%,LDH leaking and PI staining positive ratio of 2 Hz group were more than that of others.QGlu,QLacand contents ofβ-tubulin increased when the frequencu was 1 Hz.4.The electrical stimulate device run successfuly for 1 week.There is no rust on the machine and guideline.Temperature and programme displayed normally.Cells cultured in the device were alive and no bacterial contamination in 2 weeks.5.There were no significant differences of cell morphous after electrical stimulate. However,at increasing frequencies cell shortening was significantly greater in stimulated than in unstimulated myocytes.Blebs and cell debris which signified apoptosis could be observed if the stimulation was excess.Conclusion1.Cardiac myocytes kept alive from 3 to 15 days in culture with the density of 2×105/cm2 and 4×105/cm2.2.The loading device and electrical stimulate device with stable performance could be used to evaluate cell response sensitivity to mechanical stress and electrical stimulation in vitro.3.Cells had appeared slender with bright outline and regularly aligned evenly distributed porch stress circumcising line.Proper mechanical stretch stress can enhance effect on the metabolism and contents ofβ-tubulin.Changes were more obvious when strain was 8%,1 Hz. 4.We have found that regular electrical stimulation of adult ventricular myocytes in culture,so that they contract rhythmically,enhances both glucose metabolism and contents ofβ-tubulin which anent the mechanical properties of cardiomyocytes.These results suggest that regular electrical stimulation is important when studying the function of adult ventricular myocytes in culture.
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