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

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Summary

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.

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References

  • 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: 1382–1384

    Google Scholar 

  • 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 293:374–376

    Google Scholar 

  • Dean PN (1980) Dual beam sorting at Livermore. In: Laerum OD, Lindmo T, Thorud E (eds) Flow cytometry IV. Universitetsforlaget, Bergen Oslo Tromsø, pp 41–44

    Google Scholar 

  • Dean PN, Pinkel D (1978) High resolution dual laser flow cytometry. J Histochem Cytochem 26:622–627

    Google Scholar 

  • 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 2:95–96

    Google Scholar 

  • 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 13:437–447

    Google Scholar 

  • 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 72:1231–1234

    Google Scholar 

  • Gray JW, Langlois RG, Carrano AV, Burkhart-Schultz KJ, Van Dilla MA (1979) High resolution chromosome analysis: One and two parameter flow cytometry. Chromosoma 73:9–27

    Google Scholar 

  • 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 

  • Hilwig I, Gropp A (1972) Staining of constitutive heterochromatin in mammalian chromosomes with a new fluorochrome. Exp Cell Res 75:122–126

    Google Scholar 

  • Horan PK, Wheeless Jr LL (1977) Quantitative single cell analysis and sorting. Science 198:149–157

    Google Scholar 

  • Langlois RG, Carrano AV, Gray JW, Van Dilla MA (1980) Cytochemical studies of metaphase chromosomes by flow cytometry. Chromosoma 77:229–251

    Google Scholar 

  • Latt SA, Wohlleb JC (1975) Optical studies of the interaction of 33258 Hoechst with DNA, chromatin, and metaphase chromosomes. Chromosoma 52:297–316

    Google Scholar 

  • LePecq JB, Paoletti C (1967) A fluorescent complex between ethidium bromide and nucleic acids. J Mol Biol 27:87–106

    Google Scholar 

  • 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 76:5804–5808

    Google Scholar 

  • 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 60:10–15

    Google Scholar 

  • Melamed MR, Mullaney PF, Mendelsohn ML (eds) (1978) Flow cytometry and sorting. John Wiley & Sons. New York Chichester Brisbane Toronto

    Google Scholar 

  • Sawin VL, Rowley JD, Carrano AV (1979) Transcription and hybridization of 125I-cRNA from flow sorted chromosomes. Chromosoma 70:293–304

    Google Scholar 

  • Sillar R, Young B (1981) A new method for the preparation of metaphase chromosomes for flow analysis. J Histochem Cytochem 29:74–78

    Google Scholar 

  • Weissblum B, Haenssler E (1974) Fluorometric properties of the bibenzimidazole derivative Hoechst 33258, a fluorescent probe specific for AT concentration in chromosomal DNA. Chromosoma 46:255–260

    Google Scholar 

  • 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 78:7727–7731

    Google Scholar 

  • Yu L-Ch, Aten J, Gray J, Carrano AV (1981) Human chromosome isolation from short-term lymphocyte culture for flow cytometry. Nature 293:154–155

    Google Scholar 

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Work performed under the auspices of the U.S. Department of Energy by the Lawrence Livermore National Laboratory under contract number W-7405-ENG-48 with support from the Deutsche Forschungsgemeinschaft (Cr 60/3-1 and Wo 148/18)

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Cremer, C., Gray, J.W. & Ropers, H.H. Flow cytometric characterization of a Chinese hamster x man hybrid cell line retaining the human Y chromosome. Hum Genet 60, 262–266 (1982). https://doi.org/10.1007/BF00303014

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  • DOI: https://doi.org/10.1007/BF00303014

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