Summary
A complex investigation of the phenotype of a stable strain of LHC—162 cells derived from a spontaneous abortus was carried out. The karyotype, of the strain was 47,XY,+C. The extra chromosome was identified by Giemsa staining as a No. 7. The strain was subjected to cytomorphological, autoradiographic, immunochemical and virological examination in comparison with diploid cell strains. The cells of the LHC—162 strain had no oncogenic activity. Their susceptibility to the three types of poliomyelitis virus did not differ from those of diploid strains. The growth of the LHC—162 cells was poorly organized, and they had a reduced capacity for the formation of histotypical structures. There was low collagen production, poor accumulation of lipid granules, high acid phosphatase, activity, and high glycogen content. Radioautographic investigation of cell cycle revealed lengthening of the mitotic cycle in the G2, period which was twice as long as in diploid strains. Immunochemical investigation showed that fibroblasts of both trisomic and diploid strains synthesized proteins in α1 and α2 globulin zones. However, the synthesis of a protein in α1 zone was considerably more intensive and in α2 zone less intensive in the LHC—162 cells than in the diploid strain.
The stability of these phenotypic features make it reasonable to assume that they represent a biological characteristic of the LHC—162 strain. It is suggested that this characteristic totality of phenotypical features can be considered as an expression of the C7 trisomic cell's differentiation confusion. It is suggested that “the cellular syndrome”, is not due only to this chromosomal aberration but also to other chromosomal aberrations.
Zusammenfassung
Es wurde eine Komplexuntersuchung, des Phenotypus der Zellen des stabilen Stammes LHC—162 (spontaner. Abortus) durchgeführt. Der Karyotyp der Zellen dieses Stammes ist 47,XY.+C. Das überzählige Chromosom wurde durch die Giemsa-Methode als C7 identifiziert. Der Karyotyp ist im Verlauf von 62 Passagen stabil geblieben., Die cytomorphologischen, autoradiographischen, immunochemischen Untersuchungen dieses Stammes wurden im Vergleich zu diploiden Stämmen, durchgeführt. Die Zellen dieses Stammes sind nicht krebsauslösend. Die Empfindlichkeit dieses Stammes gegenüber drei Polimyelitis-Virusstämmen ist nicht anders als die Empfindlichkeit des diploiden Stammes. Das Wachstum der Zellen des Stammes LHC—162 ist wenig organisiert; sie besitzen sehr wenig, Fähigkeit, eine histotypische Struktur zu formen; sie bilden wenig Kollagen, sie speichern zu wenig Lipidgranulan; in ihnen ist der Gehalt an Glykogen und die Aktivität von Phosphotasen zu hoch. Die radiographische Untersuchung des Zellcyclus hat gezeigt, daß die Zeit des mitotischen Cyclus der G2-Periode vergrößert ist. Sie ist fast doppelt so lang wie bei dem diploiden Stamm. Bei der immunochemischen Untersuchung wurde entdeckt, daß der Stamm LHC—162 wie die diploiden Stämme Eiweiß-Komponenten in α1- und α2-Globulinzonen synthetisierte, aber der Stamm LHC—162 synthetisiert Eiweiß in der α1-Globulinzone intensiver und Eiweiß in der Globulinzone α2 schwächer als normale Zellen. Die Stabilität der, gefundenen phänotypischen Besonderheiten des Stammes LHC—162 läßt sie charakteristisch für diesen Stamm erscheinen. Diese charakteristische Gesamtheit der phänotypischen Eigenschaften kann man als Äußerung der gestörten Differenzierung der Zellen mit Trisomie C7 betrachten. Möglicherweise gibt es ähnliche “Zellsyndrome” auch für andere Chromosomenanomalien.
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References
Angell, R. R.: Cytogenetic and genetic studies in Turner's syndroms and allied conditions in man. Ph. D. Thesis, London University 1969.
Barlow, P. W.: Differential cell division in human X chromosome mosaics. Humangenetik 14, 122 (1972).
Boone, C., Chen T. R., Ruddle F: Assignment of three human genes to chromosomes (LDH-A to II, TK to 17 and LDH to 20) and evidence for translocation between human and mouse chromosomes in somatic cell hibrids. Proc. nat. Acad. Sci (Wash.) 69, 510 (1972).
Boué, J.-C., Boué, A.: Les aberrations chromosomiques dand les avortements spontanes humaine. Presse méd. 78, 14 (1970).
Boué, A., Boué J.-C., Deshayes, E.: Lignees cellulaires humaines avec aberrations chromosomiques (trisomiques, monosomiques et triploides). Stabilite caryotyque, longevite. Path. Biol. 16, 23, 1074 (1968).
Burstone, M. S.: Enzyme histohemistry and its application in the study of neoplasms. New York-London: 1962.
Carpentier, S., Lejeune, J.: Banque de cellules diploides humaines avec caryotypes anormaux. Ann. Génét. 13, 135 (1970).
Caspersson, T., Lindsten, J., Zech, L., Buckton, K. E., Price, W. H.: Four patients with trisomy 8 identified by the fluorescence and Giemsa banding technique. J. med. Genet. 9, I (1972).
Chang, T. D., Neiwozas-Late V., Uchida, J.: Selection for trisomic cells in a mosaic fibroblast culture. Cytogenetics 8, 410 (1969).
Cox, R. P.: Regulation of alkaline phosphotase in skin fibroblast cultures from patients with mongolism. Exp. Cell Res. 37, 690 (1965).
Craig-Holmes, A. P., Shaw, M. W.: Polymorphism of human constitutive heterochromatin. Science 174, 702 (1971).
Crandall, B. F., Sparkes, R. S.: Thymidine kinase activity and human chromosome No. 18. Biochem. Genet. 5, 451 (1971).
Danes, S., Bearn, A.: Hurler's syndrome. A genetic study in cell culture. J. exp. Med. 123, I (1966).
DeMars, R.: Some studies of enzymes in cultivated human cells. Nat. Cancer Inst. Monogr. 13, 181 (1964).
Edwards, J. H., Yungcken C., Rushton, D. I., Richards, S., Mittwoch, U.: Three cases of triploidy in man. Cytogenetics 6, 81 (1967).
Epifanova, O. I., Terskikh, V. V.: Method of autoradiography in the study of cell cycles. Moscow: Nauka 1969.
Fraccaro, M., Lindsten, J., Mannini, A., Scappatici., S., Tiepolo, L.: Stability of abnormal karyotypes in cell culture. Hereditas (Lund) 62, 105 (1969).
Gendon, U. Z., Marchenko, A. T., Stepanova, L. G., Andyaparidze, D. G., Dzagurov, S. G.: Studies of poliomyelitis virus — human diploid cell culture system. Proc. Simposium on human diploid cells. Zagreb, 83 (1970).
Glan, P. V., Neustroeva, V. V.: Incorporation of H3-thymidine into the cells of cultures contaminated with mycoplasma (russ.). J. Microbiol. Epidem. Immunol. (USSR) 12, 21 (1968).
Grabar, P., Williams, C. A.: Methode permettant l'etude conjugee des proprietes electrophoretiques et immunochimiques d'un melange de proteines. Application an serum. Biochim. biophys. Acta (Amst.) 10, 193 (1953).
Grouchy, J. de, Turleau, C., Léonard, C.: Etude en fluorescence d'une trisomic C mosaique, probablement 8:46,XV/47,XY, ?8+. Ann. Génét. 14, 69 (1971).
Halpern, M., Rubin, H.: Protein releised from chik embryo fibroblast in culture. Partial characterization of the protein. Cell Res. 60, 86 (1970).
Hayflick, L., Moorhead, P. S.: The serial cultivation of human diploid cells. Exp. Cell Res. 25, 585 (1961).
Hellström, K., Hagenfeld, L., Larsson, A., Lindsten, J., Sundelin, P., Tiepolo, L.: An extra C chromosome and various metabolic abnormalities in the bone marrow from a patient with reflectory sideroblastic anaemia. J. Haematol. 8, 293 (1971).
Higurashi, M., Conen, P. E.: Comparison of chromosomal, behaviour, in cultured lymphocytes and fibroblasts from patients with chromosomal disorders and controls. Cytogenetics 10, 273 (1971).
Hochwald, G., Thorbecke, G., Asotsky, R.: Sites of formation of immune globulins and of a component, of C'3 A new technique for the demonstration of the synthesis of individual serum proteins by tissues in vitro. J. exp. Med. 114, 459 (1961).
Kitchin, P. M., Shaw, M. W.: The association pattern et the Ph chromosome. Cytogenetics 10, 235 (1971).
Kuliev, A. M.: Cytogenetic investigation of spontaneous, abortions. Humangenetik 12, 275 (1971).
Kuliev, A. M.: Correlation of phenotype and karyotype anomalies in human embryogenesis (russ.). Genetics (Moscow). 8, 129 (1972).
Kuliev, A. M., Grinberg, K. N., Vasilevsky, S. S., Stepanova, L. G., Urovskaya, G.: Human cell strains with chromosome aberrations (cytogenetical, morphological and immunochemical studies) (russ.). Genetics (Moscow) 8, 146 (1972).
Laurent, C., Robert, J. M., Grambert, J., Dutrilaux, B.: Observations cliniques et cytogénétiques de deux adultes trisomiques C en mosaique. Individualisation du chromosome surnuméraire par la technique moderne de denaturation: 47,XX,?8+. Lyon méd. 226, 827 (1971).
Lejeune, J., Dutrellaux, B. Q., Rethore, M. O., Berger, R., Debray, H., Veron, P., Gone, F., Grossiord, A.: Sur trois cas de trisomie “C”. Ann. Génét. 12, 28 (1969).
Lillie, R. D.: Histopathologic technic and practical histochemistry. New York-London: McGraw-Hill Book Company 1965.
Lozzio, C. B., Lee, K. H.: Cultures of human diploid cell strains with genetic markers. 4 Intern. Congr. Human Genet., Exerpta Med., Intern. Congr. Ser., No. 233, 114 (1971).
Marinkovich, V. A., Baluda, M. A.: In vitro synthesis of gamma M-Like globulin by various chick embrionic cells. Immunology 10, 383, (1966).
Migeon, B. R., Miller, C. S.: Human-mouse somatic cell hybrids with single human chromosome (group E): Link with thymidine kinase activity. Science 162, 1005 (1968).
Mikelsaar, A. B. N., Ananiev, E. V., Gindilis, V. M.: Probable pericentric inversion in chromosome No. I in the female child. Humangenetik 9, 316 (1970).
Miller, O. J., Allderdice, P. M., Miller, D. A., Breg, W. R., Migeon, B. R.: Assignment of human thymidine kinase gene locus to chromosome 17 by identification of its distinctive quinacrine — fluorescence in man-mouse somatic hybrid. Science 173, 244 (1971).
Mittwoch, U.: DNA synthesis in cells grown, in tissue culture from patients with mongolism. In: Mongolism, 51. London: Churchil] 1967.
Mittwoch, U.: Mongolism and sex: a common problem of cell proliferation? J. med. Genet. 9 92 (1972).
Moore, G. E., Porter, J. H., Huany, C. C.: Lymphocytoid lines from persons with sex chromosome anomalies. Science 63, 59 (1969).
Nadler, H. L., Inouge, T., Hsia, D. Y. Y.: Enzymes in cultivated human fibroblasts derived from patients with autosomal trisomy syndromes. Nature (Lond.) 213, 1261 (1967).
Pears, A. G. E.: Histochemistry, London: J. a. A. Churchill Ltd. 1969.
Prokofieva-Belgolvskaja, A. A., Grinberg, K. N., Revasov, A. A., Mikelsaar, A. B. N., Kuliev, A. M.: Phenotype — karyotype correlation in chromosome deseases (russ.) Vestn. Akad. med. Nauk (in press).
Roskin, G. I.: Microscopic technique. Moscow: Sov. Nauka 1946.
Seabright, M.: A rapid banding technique for human chromosomes. Lancet 1971 II 971.
Shaw, M. W., Krooth, R. S.: Chromosomal instability in cultured fibroblasts derived from an infant with a ring chromosome. Third Intern. Congr. of Human Genetics (Abstracts) 90 (1966).
Singh, R. P., Carr, D. H.: The anatomy and histology of XO human embryos and fetuses. Ant. Res. 55, 369 (1966).
Seleznev, Yu. V.: A modified method of staining of human chromosomes after Giemsa for detection of their linear differentiation (russ.). Bull. exp. Biol. Med. (SSR) 4, 122 (1972).
Stenram, U., Stenram, L., Bergman, C.: Viral transformation of cells. Lancet 1969 I, 531.
Therkelsen, A. J.: Frequency of sex chromatin and number of nucleoli in cells grown in tissue cultured from normal human subjects and human subject with supernumerary X-chromosomes. Acta path. microbiol. scand. Section A 78, 295 (1970).
Todaro, G. J., Martin, G. M.: Increased susceptibility of Down's syndrome fibroblasts to transformation by SV40. Proc Soc. exp. 124, 1232 (1967).
Todorov, J.: Clinical laboratory investigations in pediatrics. Sofia: Med. i Physkult. 1968.
Vasileisky, S. S., Yablokova, V. I.: On differences in the beta-fetoprotein in individual human fetuses (russ.) Bull. exp. Biol. Med. (USSR) 57, 52 (1964).
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The work was supported in part by research Grants HG-70/9684148 from Human Genetics Unit WHO.
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Kuliev, A.M., Kukharenko, V.I., Grinberg, K.N. et al. Morphological, autoradiographic, immunochemical and cytochemical investigation of a cell strain with trisomy 7 from a spontaneous abortus. Hum Genet 17, 285–296 (1973). https://doi.org/10.1007/BF00273182
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DOI: https://doi.org/10.1007/BF00273182