Yeh T C, Zhang W, Ildstad S T, Ho C
Pittsburgh NMR Center for Biomedical Research, Carnegie Mellon University, PA 15213.
Magn Reson Med. 1993 Nov;30(5):617-25. doi: 10.1002/mrm.1910300513.
Isolated rat T-cells have been labeled intracellularly, using endocytosis uptake of two superparamagnetic contrast agents, AquaMag100 and BMS180549, which are both iron-oxide particles coated with dextran. No deterioration of cell proliferation response to mitogen stimulation was observed after labeling with either superparamagnetic contrast agent. AquaMag100 particles show aggregation and co-precipitation in culture media for T-cells. BMS180549 particles not only produce no observable aggregation or co-precipitation, but also have a higher efficiency for labeling T-cells than AquaMag100. The efficiency of cell labeling was determined by measuring the decrease in the spin-spin relaxation time of the water proton in cell samples containing 1 x 10(7) labeled T-cells/milliliter of 2% w/w gelatin. After optimization of the labeling procedures, a shortening of the spin-spin relaxation time by a factor of approximately 7 to 10 has been demonstrated. Under the present experimental conditions, the up-regulation of low density lipoprotein receptor does not increase the labeling efficiency by endocytosis. Our results suggest that intracellular labeling of specific cell types can be achieved with good efficiency and the labeled cells can be detected by magnetic resonance imaging in rat testicles in vivo.
使用两种超顺磁性造影剂AquaMag100和BMS180549(二者均为包裹有葡聚糖的氧化铁颗粒)通过内吞作用摄取,对分离的大鼠T细胞进行了细胞内标记。在用任何一种超顺磁性造影剂标记后,均未观察到细胞对有丝分裂原刺激的增殖反应恶化。AquaMag100颗粒在T细胞培养基中会发生聚集和共沉淀。BMS180549颗粒不仅不会产生可观察到的聚集或共沉淀,而且标记T细胞的效率比AquaMag100更高。通过测量含有1×10⁷个标记T细胞/毫升2% w/w明胶的细胞样品中水质子自旋-自旋弛豫时间的降低来确定细胞标记效率。在优化标记程序后,已证明自旋-自旋弛豫时间缩短了约7至10倍。在目前的实验条件下,低密度脂蛋白受体的上调不会通过内吞作用提高标记效率。我们的结果表明,特定细胞类型的细胞内标记可以高效实现,并且标记的细胞可以在大鼠睾丸体内通过磁共振成像进行检测。