Kuriki H, Sonta S, Murata K
Department of Clinico-Laboratory Medicine, Kansai Medical University, Osaka, Japan.
J Clin Lab Anal. 1993;7(2):119-22. doi: 10.1002/jcla.1860070210.
The effects of different swelling solutions on the univariate flow karyotype of whole chinese hamster embryo cells (WCHE K10) and cloned chinese hamster embryo fibroblast cells (CCHE 40) were compared using four methods of chromosome isolation. Chromosomes of each cell line were prepared by the Aten, Polyamine-Digitonin, Tris-Triton, and HEPES methods and analyzed with a FACStar flow cytometer. Polyamine-Digitonin and Aten methods produced the most satisfactory flow karyotype. Structurally aberrant chromosomes in the cell lines were detected in the flow karyotype as extra peaks. CCHE 40 chromosomes 1,2,X and 3q were sorted into separate tubes and reanalyzed; and the coefficient of variation of each chromosome peak was near 5%. To apply the flow karyotype analysis to clinical cytogenetics, chromosome preparation was one of the most important factor, and it is necessary to sort the chromosome peak sharply.
使用四种染色体分离方法比较了不同膨胀溶液对中国仓鼠全胚胎细胞（WCHE K10）和克隆的中国仓鼠胚胎成纤维细胞（CCHE 40）单变量流式核型的影响。通过Aten法、多胺-洋地黄皂苷法、Tris-曲拉通法和HEPES法制备每种细胞系的染色体，并用FACStar流式细胞仪进行分析。多胺-洋地黄皂苷法和Aten法产生了最令人满意的流式核型。在流式核型中，细胞系中结构异常的染色体被检测为额外的峰。将CCHE 40细胞系的1号、2号、X染色体和3号染色体长臂分选到单独的试管中并重新分析；每条染色体峰的变异系数接近5%。为了将流式核型分析应用于临床细胞遗传学，染色体标本制备是最重要的因素之一，并且有必要使染色体峰清晰分选。
