Flow karyotype analysis and sorting of the Chinese hamster chromosomes: comparing the effects of the isolation buffers.
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.
Kuriki
H
H
Department of Clinico-Laboratory Medicine, Kansai Medical University, Osaka, Japan.
Sonta
S
S
Murata
K
K
D000818:Animals; D002021:Buffers; D002460:Cell Line; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D007621:Karyotyping / Q000379:methods*
D002021:Buffers