CAJ | 학술논문

目的通过对传统乳鼠心肌细胞体外培养的条件、方法进行改进,得到纯度高、活力好、结构功能完整的心肌细胞.方法改进心室肌组织剪切方法,分别用胰酶、Ⅱ型胶原酶依次消化分离心肌组织,严格要求所需溶液pH值,差速贴壁法和化学抑制法相结合纯化心肌细胞.结果心肌细胞12h后开始贴壁生长,24h出现单个细胞自发性搏动,48 h部分心肌细胞间形成交联,72 h心肌细胞黏合成簇,96 h后细胞伪足相互接触交织成密集的网,形成放射状排列的细胞簇,可见岛屿样搏动.细胞存活率、纯度均达98％以上.结论此种方法培养的心肌细胞纯度高、活力好、结构及功能完整,是一种可靠的原代乳鼠心肌细胞培养方法.
목적 통과대전통유서심기세포체외배양적조건、방법진행개진,득도순도고、활력호、결구공능완정적심기세포.방법 개진심실기조직전절방법,분별용이매、Ⅱ형효원매의차소화분리심기조직,엄격요구소수용액pH치,차속첩벽법화화학억제법상결합순화심기세포.결과 심기세포12h후개시첩벽생장,24h출현단개세포자발성박동,48 h부분심기세포간형성교련,72 h심기세포점합성족,96 h후세포위족상호접촉교직성밀집적망,형성방사상배렬적세포족,가견도서양박동.세포존활솔、순도균체98％이상.결론 차충방법배양적심기세포순도고、활력호、결구급공능완정,시일충가고적원대유서심기세포배양방법.
Objective The present study describes an improved in vitro culture method for obtaining high purity,vital and fully functional cardiomyocytes from neonatal rat.Methods After cutting ventricular tissue with improved method,ventricular tissues were digested with low concentrations of trypsin overnight at 4 ℃,and then underwent collagenase Ⅱ digestion.Thereafter,cardiomyocytes were purified by combined differential adhesion and chemical inhibition methods.Results Adherent cardiomyocytes were seen at 12 h after culture,spontaneously beating cardiomyocytes were observed at 24 h after culture,crosslinked cardiomyocytes were found at 48 h after culture,adhesion clustered cardiomyocytes were seen at 72 h after culture,dense network formed from inter-connected was evidenced together with radial arranged cell clusters and cell pseudopodia 96 h the mutual contact woven into and formed radically ordering cell clusters and island-like beating cardiomyocytes at 96 h after culture.The cell survival rate and purity were more than 98％.Conclusion Fully functional spontaneous beating cardiomyocytes can be obtained by the use of this improved primary neonatal rat cardiomyocytes culture method.