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中文摘要：
采用改进的胶原酶灌注方法结合Percoll密度梯度离心加anti—LSEC microbeads免疫磁珠分离纯化小鼠肝窦内皮细胞（liver sinusoidal endothelial cells，LSECs），台盼蓝染色测定细胞活力，光镜下观察培养细胞的形态学变化，流式细胞仪分析其纯度，RT—PCR方法检测其新的特征性分子LSECtin基因的表达。结果表明：此方法分离的小鼠LSECs得率为（5±0．2）×10^6个／只小鼠，台盼蓝染色活力≥95％，流式细胞仪分析其纯度达到91．92％，光镜下观察细胞形态典型，RT—PCR检测到新的特征分子LSE Ctin基因的表达。本研究建立了一种小鼠LSECs的分离、纯化、培养及鉴定方法，为其功能和作用机制的深入研究打下了基础。
英文摘要：
Mouse live sinusoidal endothelial cells （LSECs） were isolated by improving the collagenase perfusion, percoll gradient isopycnic centrifugation, and purified using anti-LSEC microbeads. The viability of cells was assessed by trypan blue staining. The changes on morphology were observed under light microscope. Cells were stained with anti-LSEC-FITC antibody and analyzed by flow cytometry. The expression of LSECtin in LSECs was detected using RT-PCR. The yield of LSECs was （5±0.2）×10^6 cells/mice, vitality is greater than 95%, and the purity is more than 91.92%. The isolated LSECs have typical morphology. The expression of LSECtin could be detected in LSECs. In conclusion, we set up an effective, stable method for the isolation and cultivation, purification and identification of mouse LSECs, which is very important for studying their functions deeply.
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