大鼠原代肝细胞的培养及鉴定

目的:探讨体外大鼠肝细胞的分离培养及纯化的方法。方法:采用胶原酶Ⅳ消化组织块法分离新生大鼠肝细胞,对大鼠肝细胞进行纯化、培养,并采用免疫组织化学方法对其进行鉴定。结果:培养24 h,有部分细胞贴壁,48 h有大量细胞贴壁,但含有较多杂细胞。经过4~5次传代后,细胞形态一致,可见许多双核细胞和多核细胞。采用抗大鼠细胞角蛋白(ck-18)的特异抗体进行免疫细胞化学鉴定,经SP染色DAB显色后,阳性着色部位在胞浆呈棕黄色颗粒,阴性对照未见着色。结论:成功地体外培养了大鼠肝细胞并对其进行了鉴定,为进一步的实验研究奠定了基础。

Identification and primary culture of rat liver cells

Objective: Study of the methods of culturing and purifying of rat hepatocytes in vitro.Methods:Isolate the neonatal SD rat hepatocytes by collagenase perfusion method.These cells were cultured and purified,then identified by immunocytochemistry.Results:After culturing for 24h,there was part of adherence appeared with hepatocytes in the culture flask.more cells adhered after 48 hours,but a lot of other cells included except hepatocytes.The 4th or 5th passage cells still grew adhesively with regular cellular morphology.and binuclei or polynucleation.The cells revealed good morphologic characteristics under phase contrast microscope and showed positive in immunocytochemistry staining for an tibody ck-l8 using SP technique.Conclusion:Rat liver cells can be cultured and identified successfully with this method which may be useful in other research.