作　　者： ; ; ; ; ; ; ; ; ; ;

机构地区： 南方医科大学珠江医院

出　　处： 《南方医科大学学报》 2007年第6期824-826,共3页

摘　　要： 目的探索用半自动胰腺消化系统建立大规模人胰岛细胞纯化的可靠方法。方法使用自制的半自动人胰腺消化分离装置及胶原酶P进行人胰腺的消化分离,用COBE2991细胞分离机及HCA-Ficoll纯化人胰岛细胞。通过DTZ染色,在倒置显微镜下计数胰岛细胞的数量和纯度,用胰岛素释放试验检测胰岛细胞的分泌功能。结果消化后平均每条胰腺可平均获得(386201±78219)个胰岛细胞当量(IEQ),纯化后平均每条胰腺可获得231420±28054IEQ,平均每克胰腺组织可获得(3148±317)IEQ,纯化后胰岛细胞平均纯度为(62.81±2.68)%。纯化后的胰岛细胞对胰岛素释放刺激反应良好,高糖(16.7mmolL)时胰岛素的释放量为低糖(3.3mmolL)时的3.53倍(P≤0.001)。结论成功建立了半自动人胰岛细胞分离、纯化的方法,纯化的人胰岛细胞具有良好的生物活性。 Objective To establish an semi-automated effective method for large-scale purification of islet cells from human pancreas. Methods Human pancreas tissue was digested with collagenase P using a semi-automated pancreas-digestion system followed by purification in a HCA-Ficoll continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity was evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release assay in vitro. Results The number of the islets coil ected from each pancreas averaged 38 6201 ±78 219 islet equivalents （IEQ） before purification, and 231 420±28 054 IEQ after the purification with discontinuous gradient centrifugation. From each gram of the pancreatic tissue, 3148±317 IEQ were obtained with an average purity of （62.81 ±2.68） %. The purified islets responded well to high-concentration （16.7 mmol/L） glucose stimulation with a 2.53-fold increase of insulin secretion over the basal level （3.3 mmol/l, P〈0.001）. Conclusion The established semi-automated method can be applicable for large-scale purification of fully functional islet cells from human pancreas.

关 键 词： 胰岛细胞 移植 细胞分离

领　　域： [生物学]