CAJ | 학술논문

目的：探索和建立同步分离大鼠原代肝细胞、星状细胞、枯否细胞和肝窦内皮细胞，并进一步鉴定和培养的方法。方法联合应用原位灌注、离体灌注、梯度密度离心和差速贴壁等分离方法，获取4种大鼠肝内原代细胞，采用形态学观察、免疫荧光染色和墨汁吞噬实验等方法对所分离的各种原代细胞进行纯度鉴定。结果通过原位灌注结合离体灌注、密度梯度离心结合差速贴壁等分离手段，成功实现了大鼠原代肝细胞、星状细胞、枯否细胞以及肝窦内皮细胞的同步分离，且所得原代细胞得率、活力及纯度等指标均符合后续细胞实验要求。结论通过简易方法同步分离大鼠原代肝细胞、星状细胞、枯否细胞以及肝窦内皮细胞是可行的。
목적：탐색화건립동보분리대서원대간세포、성상세포、고부세포화간두내피세포，병진일보감정화배양적방법。방법연합응용원위관주、리체관주、제도밀도리심화차속첩벽등분리방법，획취4충대서간내원대세포，채용형태학관찰、면역형광염색화묵즙탄서실험등방법대소분리적각충원대세포진행순도감정。결과통과원위관주결합리체관주、밀도제도리심결합차속첩벽등분리수단，성공실현료대서원대간세포、성상세포、고부세포이급간두내피세포적동보분리，차소득원대세포득솔、활력급순도등지표균부합후속세포실험요구。결론통과간역방법동보분리대서원대간세포、성상세포、고부세포이급간두내피세포시가행적。
Objective To establish a method for simultaneous isolation and primary culture of rat hepatocytes, hepatic stellate cells, Kupffer's cells and hepatic sinus endothelial cells. Methods By combining in situ perfusion, in vitro perfusion, density gradient centrifugation and differential adhension, primary rat hepatocytes, hepatic stellate cells, Kupffer's cells and hepatic sinus endothelial cells were obtained. The purity of these cells were assessed with morphological observation, immunofluorescent staining and ink phagocytosis assay. Results We successfully obtained the 4 primary cells simultaneously by combining in situ perfusion with in vitro perfusion, density gradient centrifugation, and differential attachment. The cell yield rate, cell viability and purity all met requirements for the subsequent cell experiment. Conclusion The combined cell isolation and culture method is feasible to isolate primary rat hepatocytes, hepatic stellate cells, Kupffer's cells and hepatic sinus endothelial cells simultaneously.