Effects Of Collagenase With Different Activities On The Isolation Of Rat Cardiomyocytes
Keywords
Muscle Cells, Heart, Collagenases, Survival Rate, Myocardial Contraction
Abstract
¡¾Abstract Objective? ? Study the effects of collagenase with different activities on the survival rate of rat cardiomyocytes. method? ? Separated using 4 collagenases with different activities by conventional enzymatic hydrolysis? Observe the survival rate of rat cardiomyocytes immediately after isolation (time point D), 60 minutes after recalcification (time point E) and 10 minutes after electrical stimulation (time point F). IonOptix single cell? The cell edge detection system simultaneously detects the contraction amplitude of cardiomyocytes (ph), the percentage of pH in the length of a single cardiomyocyte (bl) (expressed in ph/bl), the contraction rate of cardiomyocytes (+dL/?dt) and the relaxation rate of cardiomyocytes. (-dL/dt) and other indicators. result? ? Observing the survival rate of cardiomyocytes at time points D, E and F, the results showed that the separation effect of 299U/mg collagenase was 286U/? mg, 203U/mg and 403U/mg of collagenase (P<0.05). There was no difference in the survival rate of cardiomyocytes among the 4 groups at time point D, while the survival rate decreased at time points E and F. Collected in cardiomyocytes? In the detection of systolic/diastolic function, 299U/mg collagenase has the best separation effect. The pH of the 203, 286, 299U/mg and 403U/mg groups are 0.13¡À0.05, 0.14¡À0.06, respectively? 0.14¡À0.06¦Ìm and 0.11¡À0.05¦Ìm; ph/bl are 8.8%¡À2.4%, 8.9%¡À2.3%, 9.2%¡À3.5% and 7.5%¡À2.2% respectively; +dL/dt are 1.8¡À? 0.6, 1.9¡À0.6, 2.0¡À0.7¦Ìm/s and 1.7¡À0.5¦Ìm/s, -dL/dt are 1.9¡À0.6, 2.0¡À0.7, 2.1¡À0.6¦Ìm/s and 1.7¡À0.6¦Ìm/s respectively. in conclusion? ? Adopt one? Collagenase in a certain range (203~299U/mg) is used to separate cardiomyocytes. The greater the enzyme activity, the shorter the digestion time, the higher the survival rate of cardiomyocytes and the better systolic/diastolic function. Among them? Collagenase with 299U/mg activity has the best effect. The survival rate and systolic/diastolic function of cardiomyocytes isolated from collagenase with high or low activity were significantly reduced. For further information of this article and research, feel free to contact our team for asssitance. Original research was done by Zhu Xiaoxing, Niu Xiaolin, Zhu Xiaoling
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