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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。
BACKGROUND: Peripheral blood mononuclear cell populations contain peripheral blood stem cells. The proliferation in vitro remains unclear, and in vivo transplantation lacks a good marker. OBJECTIVE: To observe the proliferation of peripheral blood mononuclear cells in stem cell culture medium with EdU as a marker, and to explore the feasibility of EdU-labeled peripheral blood stem cells. Methods: Peripheral blood mononuclear cells of New Zealand rabbits were isolated and cultured in EdU stem cell culture medium for 1-5 days. The cell morphology and cell count were observed at different time points. EdU cells were harvested for EdU staining. Test positive rate. RESULTS AND CONCLUSION: (1) Peripheral blood mononuclear cells were isolated in a round shape with clear outline. After 1 d culture, most of the cells were still suspended in culture medium, spherical or round, with cell mass , Also see a small amount of cells scattered adherent, showing triangular or polygonal; cultured 2 d, we can see more cell debris, most of the cells are still round; cultured 3-5 d, the cell debris increased, the cell volume becomes smaller , And the cell density decreased significantly. (2) The cell counts decreased gradually with the prolongation of culture time. (3) EdU red labeled cells were scattered on day 1 of culture. EdU red labeled cells increased obviously on day 2, And remained unchanged for three or four days. The number of EdU red labeled cells decreased significantly after 5 days of culture. The highest rate of EdU positive cells was (2.38 ± 0.10)% after cultured for 2 days. (4) The results showed that , The peripheral blood mononuclear cell population in the proliferation of cells is very low, EdU can be marked peripheral blood mononuclear cells have proliferative cells.