EdU标记兔外周血单个核细胞的体外增殖

发布时间：2018-05-17 03:27

本文选题：干细胞 + 细胞增殖　；参考：《中国组织工程研究》2017年09期

【摘要】：背景:外周血单个核细胞群中含有外周血干细胞,其体外增殖情况仍不清楚,体内移植也缺乏良好的标记物。目的:以EdU作为标记物,观察外周血单个核细胞在干细胞培养基中的增殖情况,以及探讨EdU标记外周血干细胞的可行性。方法:分离新西兰兔外周血单个核细胞,在加入EdU的干细胞培养液中培养1-5 d;于不同的时间点观察细胞形态和进行细胞计数;收获细胞,进行EdU染色,观察EdU阳性细胞并检测阳性率。结果与结论:(1)刚分离出的外周血单个核细胞为类圆形,轮廓清晰;经过1 d培养时,大部分细胞仍悬浮在培养液中,球形或类圆形,有细胞团出现,也见少量细胞散在贴壁,呈现三角形或多边形;培养2 d时,可见较多细胞碎片,大多数细胞仍为类圆形;培养3-5 d时,细胞碎片增多,细胞团体积变小,细胞密度明显降低;(2)随着培养时间的延长,细胞计数逐渐降低;(3)在培养1 d时,有散在EdU红色标记细胞;在培养2 d时,EdU红色标记细胞明显增多,并在培养三四天时维持不明显的变化;在培养5 d时,EdU红色标记的细胞明显减少;培养2 d时,EdU阳性细胞率最高,为(2.38±0.10)%;(4)结果表明,外周血单个核细胞群中增殖细胞率很低,EdU能标记外周血单个核细胞中有增殖能力的细胞。
[Abstract]:Background: peripheral blood mononuclear cells (PBMC) contain peripheral blood stem cells (PBSCs). Aim: to observe the proliferation of peripheral blood mononuclear cells (PBMC) in stem cell culture medium using EdU as marker and to explore the feasibility of EdU labeling peripheral blood stem cells (PBSCs). Methods: the peripheral blood mononuclear cells (PBMC) of New Zealand rabbits were isolated and cultured in stem cell culture medium supplemented with EdU for 1-5 days. Cell morphology and cell count were observed at different time points. Cells were harvested and stained with EdU. EdU positive cells were observed and the positive rate was detected. Results and conclusion: the peripheral blood mononuclear cells (PBMC) isolated from the first day of culture were round and clear, and most of the cells were still suspended in the culture medium after 1 day culture, the cells were round or round, there were cell clusters, and a few cells were scattered in the adherent to the wall. After 2 days of culture, more cell fragments were found, and most of the cells were still round. After 3 to 5 days of culture, the number of cell fragments increased, the cell mass became smaller, and the cell density decreased significantly with the prolongation of culture time. The number of EdU red labeled cells was decreased gradually at 1 day after culture, and the number of red labeled cells increased significantly at 2 days, and maintained no obvious change at 3 or 4 days after culture. The positive rate of Edu was the highest at 2 days (2.38 卤0.10). The proliferation rate of peripheral blood mononuclear cells was very low and Edu could label the proliferative cells in peripheral blood mononuclear cells.
【作者单位】：中山大学中山眼科中心眼科学国家重点实验室;
【基金】：广东省科技计划项目(2013B040200020,2013B020400003) 广州市科技计划项目(15570001)~~
【分类号】：R329.2

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