Human Genetics

, Volume 60, Issue 3, pp 262–266

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

  • C. Cremer
  • J. W. Gray
  • H. H. Ropers
Original Investigations


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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Carrano AV, Gray JW, Langlois RG, Burkhart-Schultz KJ, Van Dilla MA (1979) Measurement and purification of human chromosomes by flow cytometry and sorting. Proc Natl Acad Sci USA 76: 1382–1384Google Scholar
  2. Davies KE, Young BD, Elles RG, Hill ME, Williamson R (1981) Cloning of a representative genomic library of the human X chromosomes after sorting by flow cytometry. Nature 293:374–376Google Scholar
  3. Dean PN (1980) Dual beam sorting at Livermore. In: Laerum OD, Lindmo T, Thorud E (eds) Flow cytometry IV. Universitetsforlaget, Bergen Oslo Tromsø, pp 41–44Google Scholar
  4. Dean PN, Pinkel D (1978) High resolution dual laser flow cytometry. J Histochem Cytochem 26:622–627Google Scholar
  5. Disteche CM, Orkin SH, Lojewski A, Kunkel LM, Eisenhard M, Sahar E, Latt SA (1981) Construction of an X-enriched lambda phage library from sorted mouse chromosomes. Cytometry 2:95–96Google Scholar
  6. Gee PA, Ray M, Mohandas T, Douglas GR, Palser HR, Richardson BJ, Hamerton JL (1974) Characteristics of an HPRT-deficient Chinese hamster cell line. Cytogenet Cell Genet 13:437–447Google Scholar
  7. Gray JW, Carrano AV, Steinmetz LL, Van Dilla MA, Moore II DH, Mayall BH, Mendelsohn ML (1975) Chromosome measurement and sorting by flow systems. Proc Natl Acad Sci USA 72:1231–1234Google Scholar
  8. Gray JW, Langlois RG, Carrano AV, Burkhart-Schultz KJ, Van Dilla MA (1979) High resolution chromosome analysis: One and two parameter flow cytometry. Chromosoma 73:9–27Google Scholar
  9. Gray JW, Carrano AV, Langlois R, Lucas J, Yu L-C, Van Dilla MA (1982) Flow cytogenetics: Chromosome classification and purification by flow cytometry and sorting. In: Galjaard H (ed) The future of prenatal diagnosis. Churchill Livingstone, Edinburgh (in press)Google Scholar
  10. Hilwig I, Gropp A (1972) Staining of constitutive heterochromatin in mammalian chromosomes with a new fluorochrome. Exp Cell Res 75:122–126Google Scholar
  11. Horan PK, Wheeless Jr LL (1977) Quantitative single cell analysis and sorting. Science 198:149–157Google Scholar
  12. Langlois RG, Carrano AV, Gray JW, Van Dilla MA (1980) Cytochemical studies of metaphase chromosomes by flow cytometry. Chromosoma 77:229–251Google Scholar
  13. Latt SA, Wohlleb JC (1975) Optical studies of the interaction of 33258 Hoechst with DNA, chromatin, and metaphase chromosomes. Chromosoma 52:297–316Google Scholar
  14. LePecq JB, Paoletti C (1967) A fluorescent complex between ethidium bromide and nucleic acids. J Mol Biol 27:87–106Google Scholar
  15. Lebo RV, Carrano AV, Burkhart-Schultz K, Dozy AM, Yu L-C, Kan YW (1979) Assignment of human β-, γ-, and δ-globin genes to the short arm of chromosome 11 by chromosome sorting and DNA restriction enzyme analysis. Proc Natl Acad Sci USA 76:5804–5808Google Scholar
  16. Lebo RV, Kan YW, Cheung MC, Carrano AV, Yu L-C, Chang JC, Cordell B, Goodman HM (1982) Assigning the polymorphic human insulin gene to the short arm of chromosome 11 by chromosome sorting. Hum Genet 60:10–15Google Scholar
  17. Melamed MR, Mullaney PF, Mendelsohn ML (eds) (1978) Flow cytometry and sorting. John Wiley & Sons. New York Chichester Brisbane TorontoGoogle Scholar
  18. Sawin VL, Rowley JD, Carrano AV (1979) Transcription and hybridization of 125I-cRNA from flow sorted chromosomes. Chromosoma 70:293–304Google Scholar
  19. Sillar R, Young B (1981) A new method for the preparation of metaphase chromosomes for flow analysis. J Histochem Cytochem 29:74–78Google Scholar
  20. Weissblum B, Haenssler E (1974) Fluorometric properties of the bibenzimidazole derivative Hoechst 33258, a fluorescent probe specific for AT concentration in chromosomal DNA. Chromosoma 46:255–260Google Scholar
  21. Young BD, Ferguson-Smith MA, Sillar R, Boyd E (1981) High resolution analysis of human peripheral lymphocyte chromosomes by flow cytometry. Proc Natl Acad Sci USA 78:7727–7731Google Scholar
  22. Yu L-Ch, Aten J, Gray J, Carrano AV (1981) Human chromosome isolation from short-term lymphocyte culture for flow cytometry. Nature 293:154–155Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • C. Cremer
    • 1
  • J. W. Gray
    • 2
  • H. H. Ropers
    • 1
  1. 1.Institute of Human GeneticsUniversity of FreiburgFreiburgFederal Republic of Germany
  2. 2.Biomedical Sciences Division, Lawrence Livermore National LaboratoryUniversity of CaliforniaLivermoreUSA

Personalised recommendations