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造血系统肿瘤细胞遗传学分析的主要问题在于不能确定所得染色体来自何系细胞。作者介绍的方法能同时检测分裂细胞的核型、形态,以及确定核型异常细胞的系别。骨髓或血标本用Ficoll-Hypaque密度梯度离心,分离有核细胞,所得细胞置RPMI 1640中孵育过夜,翌日晨,加秋水仙胺(0.4μg/ml)后,孵育2—3小时,然后在2 ml细胞悬液(含细胞10~5个)内加入2 ml低渗液,置5分钟,将悬液分装在12个细胞离心容器中,400rpm离心10分钟,移细胞至截片,空气干燥过夜,以备进一步分析。涂片作Gimsa或May-Grumwald-Gimsa染色,可进行细胞学分析;苏丹黑B染色用于识别幼稚粒细胞;α-醋酸萘酯酶(ANAE)染色以鉴定单核细胞。作免疫学观察的涂片用甲醇-甲醛固定,与特异
The main problem with genetic analysis of hematopoietic tumor cells is that it is not possible to determine what lineage the resulting chromosome came from. The method described by the authors can simultaneously detect the karyotype and morphology of dividing cells and determine the karyotypic abnormal cell lineage. Bone marrow or blood samples were centrifuged with Ficoll-Hypaque density gradient and nucleated cells were isolated. The resulting cells were incubated in RPMI 1640 overnight. On the following morning, colchicine (0.4 μg/ml) was added and incubated for 2-3 hours, then in 2 Add 2 ml of hypotonic liquid to the cell suspension (containing 10 to 5 cells) for 5 minutes. Dispense the suspension in 12 cell centrifuges. Centrifuge at 400 rpm for 10 minutes. Transfer the cells to the slides and air-dry. Stay overnight for further analysis. Smears were stained with Gimsa or May-Grumwald-Gimsa for cytological analysis; Sudan Black B staining was used for identification of immature granulocytes; and alpha-naphthyl naphthalene esterase (ANAE) was stained for identification of monocytes. Immunostained smears fixed with methanol-formaldehyde and specific