大鼠胰腺星状细胞分离培养与鉴定

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。

Isolation and Identification of Rat Pancreatic Stellate Cells

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.