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1 原理将周围血(或骨髓)在适当培养液中进行短期培养,再加入植物血凝素,刺激细胞进行有丝分裂,经一定时间后加入秋水仙素以抑制丝状分裂的进行,使停止在分裂中期,最后将细胞培养物用低渗膨胀,使染色体分散展开,再进行制片染色分析。2 试剂(1)无钙镁亨氏液(Hank’s);氛化钠8g,氯化钾400mg,碳酸氢钠350mg,磷酸氢二钠152mg,无水磷酸二氢钾60mg,葡萄糖1g.0.4％/日酚红5ml,蒸溜水加至1000ml,用5.6％碳酸氢钠调pH至7.2,4℃保存。(2)双抗稀释法:①青霉素100u/ml,取青霉素G钠(钾)盐40万u加Hank’s液4ml,则每ml含10万u,用时100ml培养液内
Principle 1 The peripheral blood (or bone marrow) in the appropriate culture medium for short-term culture, then add phytohemagglutinin to stimulate cell mitosis, after a certain period of time by adding colchicine to inhibit filamentous division, stop in the split In the medium term, the cell culture is finally inflated with hypotonicity to disperse the chromosomes and perform a production staining analysis. 2 Reagents (1) Hank’s without calcium and magnesium; 8 g of sodium chloride, 400 mg of potassium chloride, 350 mg of sodium bicarbonate, 152 mg of disodium hydrogen phosphate, 60 mg of anhydrous potassium dihydrogen phosphate, 1 g of glucose 0.4% / day Phenol red 5ml, distilled water added to 1000ml, with 5.6% sodium bicarbonate to pH 7.2, stored at 4 ℃. (2) double anti-dilution method: ① penicillin 100u / ml, taking penicillin G sodium (potassium) 400000 u plus Hank’s solution 4ml, then each containing 100000 u ml, with 100ml culture medium