Flow cytometric characterization of a Chinese hamster x man hybrid cell line retaining the human Y chromosome

✍ Scribed by C. Cremer; J. W. Gray; H. H. Ropers

• Publisher: Springer
• Year: 1982
• Tongue: English
• Weight: 523 KB
• Volume: 60
• Category: Article
• ISSN: 0340-6717
• DOI: 10.1007/bf00303014

✦ Synopsis

A Chinese hamster X man hybrid cell line (CH-Y-VII) was established which retains a free human Y chromosome. Exponentially growing CH-Y-VII cells were arrested with colcemid; metaphase chromosomes were isolated and stained with 33258 Hoechst (HO) plus Chromomycin A3 (CA3), or with ethidium bromide (EB). The HO/CA3-stained chromosomes were measured in a dual beam flow cytometer, and bivariate HO/CA3 flow karyotypes and univariate HO and CA3 flow karyotypes were established. EB-stained chromosomes were analyzed in a modified Becton Dickinson FACS-Sorter. For all three stains used, the human Y chromosome forms a separate peak in univariate flow karyotypes; the optimum resolution was obtained for the HO distribution. In the bivariate HO/CA3 flow karyotype, the peak for the human Y chromosome is completely separated from the Chinese hamster chromosomes.