Summary
When PHA stimulated human lymphoid cells are allowed to proliferate in vitro and chromosomal slides are prepared from the culture after 48,72, 96,120, and 144h of growth, gradual changes in chromosome morphology can be observed after traditional Giemsa staining of the slides. Keeping culture conditions, colcemid exposure, and fixation procedure constant for all samples, it is found that average chromosome length decreases with increasing culture time. A shift from high frequencies of subbanded chromosomes (sample 48 h and sample 72 h) to high frequencies of unbanded and G banded chromosomes (sample 120 h and sample 144 h) takes place simultaneously with the general compaction of the chromosomes. Examination of trypsin-induced G bands as well as examination of untreated G banded chromosomes from all samples clearly indicate that the basic G band pattern is not altered during proliferation and differentiation, although the progressive compaction of the chromosome observed with increasing culture time results in a phenomenon similar to that observed during mitosis, where the compact late metaphase chromosome after trypsin treatment exhibits fewer but more prominent bands than the prophase/prometaphase chromosomes. Thus the progressive compaction of metaphase chromosomes observed during in vitro aging seems to resemble the condensation processes during the G2 phase and mitosis.
It has been suggested that the chromomeres serve as centers for chromosome condensation during mitosis, probably mediated by a sulphydryl-disulphide transition in chromosomal proteins. The data presented here further suggest that the chromomeres may also serve as centers for chromosomal differentiation, presumable by a mechanism similar to that acting during chromosome condensation in mitosis.
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References
Andersen, O., Rønne, M.: Effect of histidinol anparafluorophenylalanine on metaphase chromosome structure in human lymphoid cells. Hereditas 88, 197–201 (1978)
Beek, B., Obe, G.: The human leucocyte test system IV. The RNA-synthesis pattern in the first G1-phase of the cell cycle after stimulation with PHA. Mutat. Res. 29, 165–168 (1975)
Beek, B., Obe, G.: Sister chromatid exchanges in human leukocyte chromosomes: Spontaneous and induced frequencies in early- and late-proliferating cells in vitro. Hum. Genet. 49, 51–61 (1979)
Bostock, C. J., Summer, A. T.: Changes in chromosomes during mitosis. In: The eukaryotic chromosome, 287–292. Amsterdam: Elservier North Holland Biochemical Press 19
Brown, S. W.: Heterochromatin. Science N.Y. 151, 417–425 (1966)
Comings, D. E.: Mechanisms of chromosome banding and implications for chromosome structure. Ann. Rev. Genet. 12, 25–46 (1978)
Crossen, P. E., Morgan, W. F.: Analysis of human lymphocyte cell cycle time in culture measured by sister chromatid differential staining. Exp. Cell Res. 104, 453–457 (1977)
Dudin, G., Beek, B., Obe, G.: The human leucocyte test system I. DNA synthesis and mitosis in PHA-stimulated 2-day cultures. Mutat. Res. 23, 279–281 (1974)
Goodman, L. A.: Kolmogorov-Smirnov tests for psychological research. Psychol. Bull. 51, 160–168 (1954)
Hoo, J. J., Parslow, M. I.: Relation between the SCE points and the DNA replication bands. Chromosoma 73, 67–74 (1979)
Korenberg, J. R., Freedlander, E. F.: Giemsa technique for the detection of sister chromatid exchanges. Chromosoma 48, 355–360 (1974)
Latt, S. A.: Microfluorometric detection of deoxyribonucleic acid replication in human metaphase chromosomes. Proc. Natl. Acad. Sci. (USA) 70, 3395–3399 (1973)
Maur, P. auf der, Berlincourt-Böhni, K.: Human lymphocyte cell cycle: Studies with the use of BrUdR. Hum. Genet. 49, 209–215 (1979)
Ohnuki, Y.: Structure of chromosomes. I. Morphological studies of the spiral structure of human somatic chromosomes. Chromosoma 25, 402–428 (1968)
Oppenheim, J. J., Rosenstreich, D. L.: Signals regulating in vitro activation of lymphocytes. Proc. Allergy 20, 65–194 (1976)
Paris Conference 1971: Standardization in human cytogenetics birth defects. Original Article Series 8, 7. New York: The National Foundation 1972
Perry, P., Wolff, S.: New Giemsa method for the differential staining of sister chromatids. Nature 251, 156–158 (1974)
Rao, P. N.: Premature chromosome condensation and the fine structure of chromosomes. In: Chromosomes in biology and medicine (J. J. Yunis, ed.), pp. 205–231. New York, London: Academic Press 1977
Rønne, M.: In vitro induction of G bands in human chromosomes. Hereditas 85, 81–84 (1977a)
Rønne, M.: In vitro induction of G bands with cycloheximide. Hereditas 86, 107–110 (1977b)
Rønne, M.: In vitro induction of uncoiled chromosomes. Hereditas 86, 111–114 (1977c)
Rønne, M.: Induction of uncoiled chromosomes with RNA'se. Hereditas 86, 245–250 (1977d)
Rønne, M.: Human cells in suspension. I. Human lymphoid and granulopoietic cells in primary and secondary cultures. The effects of in vitro induction and prolonged metanol acetic acid fixation on metaphase chromosome structure. Cytobios 22, 191–211 (1978)
Rønne, M.: The effect of in vitro dithioerythritol exposure on human metaphase chromosome structure. Hereditas 91, 105–109 (1979)
Rønne, M., Andersen, C.: In vitro induction of uncoiled chromosomes with inhibitors of RNA and protein synthesis. Cytobios 21, 7–24 (1978)
Rønne, M., Boye, H. A., Sandermann, J.: A mounting medium for banded chromosomes. Hereditas 86, 155–158 (1977)
Rønne, M., Andersen, O., Erlandsen, M.: Effect of colcemid exposure and methanol acetic acid fixation on human metaphase chromosome structure. Hereditas 90, 195–201 (1979a)
Rønne, M., Nielsen, K. V., Erlandsen, M.: Effect of controlled colcemid exposure on human metaphase chromosome structure. Hereditas 91, 49–52 (1979b)
Schneider, E. L., Monticone, R. E.: Aging and sister chromatid exchange: II. The effect of the in vitro passage level of human fetal lung fibroblasts on baseline and mutageninduced sister chromatid exchange frequencies. Exp. Cell Res. 115, 269–276 (1978)
Schor, S. L., Johnson, R. T., Waldren, C. A.: Change in the organization of chromosomes during the cell cycle: Response to ultraviolet light. J. Cell Sci. 17, 539–565 (1975)
Siegel, S.: Nonparametric statistics, for the behavioral science, pp. 127–136. New York: McGraw-Hill Book Company Inc., 1956
Sperling, K., Rao, P. N.: Mammalian cell fusion. V. Replication behaviour of heterochromatin as observed by premature chromosome condensation. Chromosoma 45, 121–131 (1974)
Sumner, A. T.: Involvement of protein disulphides and sulphydryls in chromosome banding. Exp. Cell Res. 83, 438–442 (1973)
Sumner, A. T.: Banding as a level of chromosome organization. In: Current chromosome research (K. Jones, P. E. Brandham, eds.), pp. 17–22. Amsterdam: Elsevier, North-Holland Biochemical Press (1976)
Takayama, S.: Configurational changes in chromatids from helical to banded structures. Chromosoma 56, 47–54 (1977)
Tice, R., Schneider, E. L., Rary, J. M.: The utilization of bromodeoxyuridine incorporation into DNA for the analysis of cellular kinetics. Exp. Cell Res. 102, 232–236 (1976)
Wang, H. C., Fedorff, S.: Banding in human chromosomes treated with trypsin. Nature New Biol. 235, 52–53 (1972)
Yunis, J. J., Sawyer, J. R., Ball, D. W.: The characterization of high-resolution G-banded chromosomes of man. Chromosoma 67, 293–307 (1978)
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Rønne, M. Human cells in suspension 2. Hum Genet 54, 55–62 (1980). https://doi.org/10.1007/BF00279049
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DOI: https://doi.org/10.1007/BF00279049