【摘要】：目的研究一種簡(jiǎn)單、高效、實(shí)用并且穩(wěn)定的SD大鼠肝臟原代枯否細(xì)胞(KCs)的提取及培養(yǎng)方法。方法選擇體質(zhì)量200 g左右的健康SD雄性大鼠,采用0.05%IV型膠原酶在體原位灌注法結(jié)合剪碎法,37℃水浴振蕩20 min,經(jīng)30%和60%Percoll密度梯度離心后,再經(jīng)40%Percoll離心及貼壁法純化提取KCs。倒置顯微鏡下觀察細(xì)胞形態(tài),并用臺(tái)盼藍(lán)染色實(shí)驗(yàn)鑒定細(xì)胞活力,吞墨實(shí)驗(yàn)觀察細(xì)胞吞噬功能及流式細(xì)胞術(shù)鑒定細(xì)胞純度。結(jié)果每只大鼠平均肝臟KCs的獲得量為(1.00±0.20)×107(n=20),細(xì)胞活力為(92.34±0.15)%,細(xì)胞純度均在(93.56±0.24)%。結(jié)論改良的SD大鼠肝臟KCs提取方法操作簡(jiǎn)單,效果可靠,可為今后KCs的研究奠定基礎(chǔ)。
[Abstract]:Aim to study a simple, efficient, practical and stable method for the extraction and culture of (KCs) from primary Kupffer cells of SD rat liver. Methods healthy male SD rats, weighing about 200g, were injected with 0.05% type IV collagenase in situ combined with shear method. After 30% and 60%Percoll density gradient centrifugation, 20 min, oscillated in water bath at 37 鈩，

本文編號(hào)：2471835