大鼠肝脏原代Kupffer细胞的分离培养

Isolation and Purification of Primary Kupffer Cells from Rat Liver

目的建立稳定而有效的大鼠肝脏Kupffer细胞分离、纯化的实验方法。方法用无Ca2+灌注液和浓度为0.05%的胶原酶灌注液进行肝脏的灌注及消化,用Percoll密度梯度离心法进行Kupffer细胞的分离、纯化,并鉴定大鼠肝脏原代Kupffer细胞。结果分离培养后得到纯化的原代Kupffer细胞,生长状态良好,DAB染色实验证实经大鼠肝脏灌流得到稳定生长的原代Kupffer细胞。结论通过大鼠肝脏灌注建立了有效的大鼠肝脏Kupffer细胞分离、纯化的实验方法。

Objective To explore a stable and effective method for isolation and purification of Kupffer cells from rat liver. Methods Rat liver was perfused and digested with perfusate without calcium and 0.05% collagenase perfusate. Percoll density gradient centrifugation was performed to isolate and purificate Kupffer cells. Results DAB staining method proved that primary Kupffer cells were obiained by perfusion of rat live and the cells were in good condition. Conclusion A stable and effective method for theisolation and purification of Kupffer cells from rat liver was explored through perfusion of rat liver.