CAJ | 학술논문

目的：分离培养大鼠胰腺星状细胞,为体外研究胰腺纤维化提供细胞模型。方法：采用酶消化结合Nycodenz密度梯度离心法分离胰腺星状细胞,通过倒置显微镜观察细胞形态，油红O染色和免疫细胞化学染色desmin，α-SMA进行细胞鉴定，并绘制传代后细胞生长曲线。结果：原代培养的大鼠胰腺星状细胞呈星形或梭形生长；原代胰腺星状细胞油红O染色胞浆均可见红色密集脂滴，直至传代后消失；原代胰腺星状细胞培养48 h后，desmin表达开始减弱，α-SMA表达开始增强；传代后desmin基本不表达，α-SMA阳性表达100%；传代后第3 d~5 d的胰腺星状细胞处于快速增殖阶段。结论：酶消化结合Nycodenz密度梯度离心法分离培养大鼠胰腺星状细胞纯度高，重复性好，可以满足体外研究的需要。
목적：분리배양대서이선성상세포,위체외연구이선섬유화제공세포모형。방법：채용매소화결합Nycodenz밀도제도리심법분리이선성상세포,통과도치현미경관찰세포형태，유홍O염색화면역세포화학염색desmin，α-SMA진행세포감정，병회제전대후세포생장곡선。결과：원대배양적대서이선성상세포정성형혹사형생장；원대이선성상세포유홍O염색포장균가견홍색밀집지적，직지전대후소실；원대이선성상세포배양48 h후，desmin표체개시감약，α-SMA표체개시증강；전대후desmin기본불표체，α-SMA양성표체100%；전대후제3 d~5 d적이선성상세포처우쾌속증식계단。결론：매소화결합Nycodenz밀도제도리심법분리배양대서이선성상세포순도고，중복성호，가이만족체외연구적수요。
Objective To establish a method for isolation and culture of rat pancreatic stellate cell (PSC). Methods PSCs were isolated by enzyme digestive method combined Nycodenz discontinuous density gradi-ent centrifugation. The cell morphology was observed by inversion microscope. Oil red O staining and immunos-taining of desmin and α-SMA were performed to identify PSC. The growth curve was drawn to understand the characteristics of PSC. Results The primary PSCs were star or spindle shape during culture . There were nu-merous lipid drops in cytoplasm of primary PSC and the lipid drops disappeared after passage. After 48 hours, the primary PSCs were devoid of desmin and positive for α-SMA. There was no expression of desmin and 100%expression of α-SMA after passage. The fast proliferation period of passage PSC was from the third day to the fifth day. Conclusion PSCs were successfully isolated by enzyme digestive method combined Nycodenz dis-continuous density gradient centrifugation and this method could meet the demand for research of PSC in vitro.