CAJ | 학술논문

背景：大鼠心肌细胞原代培养技术日渐成熟，但小鼠心肌细胞在同样实验条件下不易获得，而小鼠基因组与人类基因组有更多相似之处，具有更高的研究价值。 　　目的：改进小鼠乳鼠原代心肌细胞的培养方法，得到纯度高、活力强、保持心肌细胞原有结构和功能的心肌细胞。 　　方法：将小鼠心肌组织利用酶消方法分离为单个的心肌细胞，实验步骤中将经典的胰酶、胶原蛋白酶混合酶消法分解开，先用胰酶消化心肌组织，使心肌组织变得松散，再用胶原蛋白酶，作用于细胞间质的胶原纤维，使心肌组织分离为单个的心肌细胞。调整胰酶和Ⅱ型胶原蛋白酶的浓度和作用时间，严格控制试剂pH值和各步骤的温度。 　　结果与结论：心肌细胞在接种24 h后开始贴壁，48 h后可观察到部分心肌细胞出现自主搏动，72 h后彼此形成交联，96 h后可观察到心肌细胞形成细胞簇，并出现一致性搏动。心肌细胞的存活率和纯度均达95%以上。结果证实，实验采用改良方法成功培养出小鼠乳鼠心肌细胞，并且保留心肌细胞的结构和功能，纯度和成活率高，是可行的培养方法。
배경：대서심기세포원대배양기술일점성숙，단소서심기세포재동양실험조건하불역획득，이소서기인조여인류기인조유경다상사지처，구유경고적연구개치。 　　목적：개진소서유서원대심기세포적배양방법，득도순도고、활력강、보지심기세포원유결구화공능적심기세포。 　　방법：장소서심기조직이용매소방법분리위단개적심기세포，실험보취중장경전적이매、효원단백매혼합매소법분해개，선용이매소화심기조직，사심기조직변득송산，재용효원단백매，작용우세포간질적효원섬유，사심기조직분리위단개적심기세포。조정이매화Ⅱ형효원단백매적농도화작용시간，엄격공제시제pH치화각보취적온도。 　　결과여결론：심기세포재접충24 h후개시첩벽，48 h후가관찰도부분심기세포출현자주박동，72 h후피차형성교련，96 h후가관찰도심기세포형성세포족，병출현일치성박동。심기세포적존활솔화순도균체95%이상。결과증실，실험채용개량방법성공배양출소서유서심기세포，병차보류심기세포적결구화공능，순도화성활솔고，시가행적배양방법。
BACKGROUND:A lot of work has been carried out on the development of the primary cultured rat myocardial cel s at home and abroad. The primary culture technology of rat myocardial cel s becomes more mature, but myocardial cel s from neonatal mice are not easy to be obtained under the same experimental conditions. The mouse genome has more similarities with the human genome, which has a higher research value. OBJECTIVE:To improve the primary culture method of neonatal mouse myocardial cel s, and to obtain myocardial cel s with high purity, vitality and original structure and function. METHODS:The mouse cardiac tissues were treated using an enzyme digestion method to isolate isolated single myocardial cel s:first, the cardiac tissues were digested using trypsin, and then col agenous fibers were treated with col agenase to isolate single myocardial cel s. The concentration and action time of trypsin and type II col agenase were adjusted, and the pH values of reagents and temperature of each step were strictly control ed. RESULTS AND CONCLUSION:At 24 hours after inoculation, the myocardial cel s began to be adherent;at 48 hours, independent pulsation of myocardial cel s could be observed;at 72 hours, myocardial cel s were cross-linked;and at 96 hours, myocardial cel s formed cel clusters and presented with consistent beating. The survival rate and purity of myocardial cel s were both over 95%. This modified method could successful y culture myocardial cel s with high purity and viablility from neonatal mice, and the structure and function of myocardial cel s could be retained. Therefore, it is a feasible culture method.