Abstract
The use of new cell combinations to study genetic interaction between somatic cells (e.g., fibroblast heterokaryons, hybrids, and cybrids) is hampered by the fact that no selection procedures are available for the short-term isolation of the specific fusion products. Moreover, the techniques for the isolation of cytoplasts (anucleate cells) and karyoplasts (minicells) [for a review see Ringertz and Savage (1976)] yield fractions which are contaminated considerably with nonenucleated cells. The use of these impure fractions as a fusing partner may give rise to difficulties in the interpretation of the results.
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References
Arndt-Jovin, D. J., and Jovin, T. M., 1977, Analysis and sorting of living cells according to deoxyribonucleic acid content, J. Histochem. Cytochem. 25:585–589.
Bootsma, D., Mulder, M. P., Pot, F., and Cohen, J. A., 1970, Different inheritent levels of DNA repair replication in xeroderma pigmentosum cell strains after exposure to ultraviolet irradiation, Mutat. Res. 9:507–516.
Bryant, E. M., Crouch, E., Bornstein, P., Martin, G. M., Johnston, P., and Hoehn, H., 1978, Regulaton of growth and gene activity in euploid hybrids between human neonatalfibroblasts and epitheloid amniotic fluid cells, Am. J. Hum. Genet. 30:392–405.
Chang, P. L., Joubert, G. I., and Davidson, R. G., 1979, Non-selective isolation of human somatic cell hybrids by unit-gravity sedimentation, Nature 278:168–170.
Corsaro, C. M., and Migeon, B. R., 1978a, Gene expression in euploid human hybrid cells: Ouabain resistance is codominant, Somat. Cell Genet. 4:531–540.
Corsaro, C. M., and Migeon, B. R., 1978b, Effect of intercellular communication on the selection of intraspecific human hybrids in HAT and ouabain, Somat. Cell Genet. 4:541–551.
Dean, P. N., and Pinkel, D., 1978, High resolution dual laser flow cytometry, J. Histochem. Cytochem. 26:622–627.
DeJosselin de Jong, J. E., Jongkind, J. F., and Ywema, H. R., 1980, A scanning inverted microfluorimeter with electronic shutter control for automatic measurements in micro-test plates, Anal. Biochem. 102:120–125.
De Weerd-Kastelein, E. A., Keizer, W., and Bootsma, D., 1972, Genetic heterogeneity of xeroderma pigmentosum demonstrated by somatic cell hybridization, Nature New Biol. 238:80–83.
De Wit-Verbeek, H. A., Hoogeveen, A., and Galjaard, H., 1978, Complementation studies with enucleated fibroblasts from different variants of β-galactosidase deficiency, Exp. Cell Res. 113:215–218.
Ege, T., and Ringertz, N. R., 1975, Viability of cells reconstituted by virus-induced fusion of minicells with anucleate cells, Exp. Cell Res. 94:469–473.
Galjaard, H., van Hoogstraten, J. J., De Josselin de Jong, J. E., and Mulder, M. P., 1974a, Methodology of the quantitative cytochemical analysis of single or small numbers of cultured cells, Histochem. J. 6:409–429.
Galjaard, H., Hoogeveen, A., Keijzer, W., De Wit-Verbeek, E., and Vlek-Noot, C., 1974b, The use of quantitative cytochemical analysis in rapid prenatal detection and somatic cell genetic studies of metabolic diseases, Histochem. J. 6:491–509.
Glick, D., 1961, Quantitative Chemical Techniques of Histo and Cytochemistry, John Wiley and Sons, New York.
Hales, A., 1977, A procedure for the fusion of cells in suspension by means of polyethylene glycol, Somat. Cell Genet. 3:227–230.
Harris, H., and Watkins, J. F., 1965, Hybrid cells derived from mouse and man, Nature 205:640–646.
Hoehn, H., Bryant, E. M., Johnston, P., Norwood, T. H., and Martin, G. M., 1975, Non-selective isolation-, stability and longevity of hybrids between normal human somatic cells, Nature 258:608–610.
Hoehn, H., Bryant, E. M., and Martin, G. M., 1978, The replicative life spans of euploid hybrids derived from short-lived and long-lived human skin fibroblast cultures, Cytogenet. Cell Genet. 21:282–295.
Hulett, H. R., Bonner, W. A., Sweet, R. G., and Herzenberg, L. A., 1973, Development and application of a rapid cell sorter, Clin. Chem. 19:813–816.
Jongkind, J. F., Ploem, J. S., Reuser, A. J. J., and Galjaard, H., 1974, Enzyme assays at the single cell level using a new type of microfluorimeter, Histochemistry 40:221–229.
Jongkind, J. F., Verkerk, A., and Tanke, H., 1979, Isolation of human fibroblast heterokaryons with two-colour flow sorting (FACS II), Exp. Cell Res. 120:444–448.
Jongkind, J. F., Verkerk, A., Schaap, G. H., and Galjaard, H., 1980, Non-selective isolation of fibroblast-cybrids by flow-sorting, Exp. Cell Res. 130:481–484.
Jongkind, J. F., Verkerk, A., Schaap, G. H., and Galjaard, H., 1981, Flow-sorting in studies on metabolic and genetic interaction between human fibroblasts, Acta Path. Microhiol. Scand. (A) Suppl. 274:164–169.
Jongkind, J. F., Verkerk, A., Visser, W. J., and Van Dongen, J. M., 1982, Isolation of autofluorescent “aged” human fibroblasts by flow sorting. Morphology, enzyme activity and proliferative capacity, Exp. Cell Res., (in press).
Jovin, T. M., and Arndt-Jovin, D. J., 1980, Cell separation, Trends Biochem. Sci. 5:214–219.
Keller, P. M., Person, S., and Snipes, W., 1977, A fluorescent enhancement assay of cell fusion, J. Cell Sci. 28:167–177.
Latt, S. A., and Stetten, G., 1976, Spectral studies on 33258 Hoechst and related bisbenzimidazole dyes useful for fluorescent detection of desoxyribonuceic acid synthesis, J. Histochem. Cytochem. 24:24–33.
Latt, S. A., and Wohlleb, J. C., 1975, Optical studies on the interaction of 33258 Hoechst with DNA, chromatin and metaphase chromosomes, Chromosoma 52:297–316.
Loewe, H., and Urbanietz, J., 1974, Basisch substituierte 2,6-Bis-benzimidanolderivate, eine neue chemotherapeutisch aktive Körperklasse, Arzneim. Forsch. 24:1927–1933.
Lowry, O. H., and Passonneau, J. V., 1972, A Flexible System of Enzymatic Analysis, Academic Press, New York.
Migeon, B. R., Norum, R. A., and Corsaro, C. M., 1974, Isolation and analysis of somatic hybrids derived from two human diploid cells, Proc. Natl. Acad. Sci. USA 71:937–941.
Nadler, H. L., Chacko, C. M., and Rachmeler, M., 1970, Interallelic complementation in hybrid cells derived from human diploid strains deficient in galactose-1-phosphate uridyl transferase activity, Proc. Natl. Acad. Sci. USA 67:976–982.
Ohkuma, S., and Poole, B., 1978, Fluorescence probe measurement of the intra lysosomal pH in living cells and the perturbation of pH by various agents, Proc. Natl. Acad. Sci. USA 75:3327–3331.
Parks, D. R., Bryan, V. M., Oi, V. T., and Herzenberg, L. A., 1979, Antigen-specific identification and cloning of hybridomas with a fluorescence-activated cell sorter, Proc. Natl. Acad. Sci. USA 76:1962–1966.
Ringertz, N. R., and Savage, R. E, 1976, Cell Hybrids, Academic Press, New York.
Rossilet, A., and Ruch, F., 1968, Cytofluorimetric determination of lysine with dansylchloride, J. Histochem. Cytochem. 16:459–466.
Rotman, B., and Papermaster, B. W., 1966, Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters, Proc. Natl. Acad. Sci. USA 55:134–141.
Schaap, G. H., Van der Kamp, A.W. M, Ory, F. G., and Jongkind, J.F., 1979, Isolation of anucleate cells using a fluorescence activated cell sorter (FACS II), Exp. Cell Res. 122:422–426.
Schaap, G. H., Verkerk, A., Van der Kamp, A. W. M., Ory, F. G., and Jongkind, J. F., 1981, Separation of cytoplasts from nucleated cells by flow sorting using Hoechst 33342, Acta Patho. Microbiol. Scand. [A] Suppl. 274:159–163.
Siniscalco, M., Klinger, H. P., Eagle, H., Koprowski, H., Fujimoto, W. Y., and Seegmiller, J. E., 1969, Evidence for intergenic complementation in hybrid cells derived from two human diploid strains each carrying an X-linked mutation, Proc. Natl. Acad. Sci. USA 62:793–799.
Udenfriend, S., Stein, S., Böhlen, P., Dairman, W., Leimgruber, W., and Weigele, M., 1972, Fluorescamine: A reagent for assay of aminoacids, peptides, proteins, and primary amines in the picomole range, Science 178:871–872.
Van Diggelen, O. P., Galjaard, H., Sinnott, M. L., and Smith, P. J., 1980, Specific inactivation of lysosomal glycosidases in living fibroblasts by the corresponding glycosylmethyl-p-nitrophenyltriazenes, Biochem. J. 188:337–343.
Veomett, G., Prescott, D. M., Shay, J., and Porter, K. R., 1974, Reconstruction of mammalian cells from nuclear and cytoplasmic components separated by treatment with cytochalasin B, Proc. Natl. Acad. Sci. USA 71:1999–2002.
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Jongkind, J.F., Verkerk, A. (1982). Nonselective Isolation of Fibroblast Heterokaryons, Hybrids, and Cybrids by Flow Sorting. In: Shay, J.W. (eds) Techniques in Somatic Cell Genetics. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-4271-7_7
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DOI: https://doi.org/10.1007/978-1-4684-4271-7_7
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