大鼠Kupffer细胞分离、培养及永生化实现

Isolation, culture and immortalization of Kupffer cells in rats

枯否细胞(Kupffer Cells,KC)是肝脏免疫防御的第一道防线,其与多种肝脏疾病密切关联。Kupffer细胞分离纯化技术要求较高,且该细胞分化增殖能力极弱,成为开展大规模细胞实验的主要障碍。课题组简化了Kupffer细胞分离纯化步骤,构PDS152_p Lenti-GFP-IRES-SV40LT表达载体,包装形成慢病毒颗粒侵染Kupffer细胞,侵染后细胞生长速度变快,能多次传代,细胞状态稳定,巨噬细胞特异性标志F4/80呈阳性表达。将SV40LT片段导入Kupffer细胞可实现细胞永生化,且转染后的kupffer细胞仍维持巨噬细胞的功能特性。

Kupffer cells(KC)is the first line of defense for the liver's immune defense,and Kupffer cells are closely related to a variety of liver diseases.Kupffer cells have many obstacles to carry out on a large scale,because of the high requirement for isolation and purification of Kupffer cells and the ability of Kupffer cell differentiation and proliferation is very weak.Our group simplified the steps of isolation and purification of Kupffer cells,and established the carrier of PDS152_pLenti-GFP-IRES-SV40LT expression.Packaging of lentivirus particles to infect Kupffer cells,we can find the growth rate of cells becomes faster after infection,multiple passages,and cell state stability.also macrophage specific marker F4/80positive expression.The introduction of SV40LT fragments into Kupffer cells can achieve immortalization of cells and the transfected Kupffer cells still maintain the functional properties of macrophages.