Hill R S, Kennedy M, Mackinder C
Pathology. 1978 Jan;10(1):69-75. doi: 10.3109/00313027809063481.
Human peripheral blood leucocytes (neutrophil-rich) were collected either with heparin or acid citrate dextrose, frozen with dimethyl sulphoxide at a controlled rate, stored in liquid nitrogen at--196 degrees C and reconstituted with a solution containing dextran. After reconstitution, 20.2% of cells (in absolute numbers 1 in 5 fresh cells) showed a strongly positive nitroblue tetrazolium (NBT) reaction. The quantitative NBT test confirmed the synthesis of formazan/10(6) reconstituted neutrophilsa s15% of the fresh capacity. A slow titration reconstitution method for cells did not improve the functional capacity of thawed leucocytes as judged by the NBT test. When comparing anticoagulants, heparin increased the post-reconstitution cell yields after freezing and increased the absolute number of reconstituted cells capable of developing a positive NBT reaction.
采集富含中性粒细胞的人外周血白细胞,使用肝素或酸性枸橼酸盐葡萄糖抗凝,以可控速率用二甲基亚砜冷冻,在-196℃液氮中储存,并用含葡聚糖的溶液复溶。复溶后,20.2%的细胞(绝对数量为每5个新鲜细胞中有1个)呈现强阳性硝基蓝四氮唑(NBT)反应。定量NBT试验证实,每10⁶个复溶中性粒细胞中azan的合成量为新鲜细胞能力的15%。根据NBT试验判断,细胞的缓慢滴定复溶方法并未改善解冻白细胞的功能能力。比较抗凝剂时,肝素可提高冷冻后复溶的细胞产量,并增加能够产生阳性NBT反应的复溶细胞的绝对数量。