Summary
3H-thymidine labelling studies and a computer simulation have been employed to assess proliferative status and cellular organisation in colonic explants maintained in culture for 5 to 7 days. The one-hour flash labelling index (Is) for crypts within the middle region of expiants (5.2%) was considerably lower than that observed in vivo (8.8%). Crypt length and the distribution of labelled cells appeared similar for both situations. A computer simulation program for crypt-cell proliferation was devised, facilitating the modulation of a number of parameters including the cell-cycle time (Tc) and its component phases, the cut-off position, and cell loss at mitosis. This simulation was employed to model continuous labelling (72 h) data obtained in vitro and provided an estimate of various kinetic parameters. Data for the middle region of expiants was fitted with a Tc of 62 h, an S phase of 8 h and a cell loss factor (20%) which was consistent with histological findings. A fit to the experimental data obtained in vitro could be achieved by a model based upon a mode of cellular organisation known to occur within crypts in vivo. Therefore in vitro, the dynamic processes of crypt-cell proliferation and migration appear to be organised in the same manner as seen in vivo.
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References
Autrup H (1980) Explant culture of human colon. In: Harris CC, Trump BF, Stoner GB (eds) Methods in cell biology, vol 21, normal human tissue and cell culture B. Academic Press, New York, pp 385–401
Autrup H, Harris CC, Stoner GD, Jesudason ML, Trump BF (1977) Binding of chemical carcinogens to macromolecules in cultured human colon. J Natl Cancer Inst 59:351–354
Autrup H, Barrett LA, Jackson FE, Jesudason ML, Stoner G, Phelps P, Trump BF, Harris CC (1978a) Expiant culture of human colon. Gastroenterology 74:1248–1257
Autrup H, Stoner GD, Jackson F, Harris CC, Shamsuddin AKM, Barrett LA, Trump BF (1978b) Expiant culture of rat colon: a model system for studying metabolism of chemical carcinogens. In Vitro 14:868–877
Cairnie AB, Lamerton LF, Steel GG (1965) Cell proliferation studies in the epithelium of the rat. II. Theoretical aspects. Expl Cell Res 39:539–553
Chang WWL, Nadler NJ (1975) Renewal of the epithelium in the descending colon of the mouse. IV. Cell population kinetics of vacuolated-columnar, and mucous cells. Am J Anat 144:39–56
Cheng H, Leblond CP (1974) Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine.V. Unitarian theory of the origin of the four epithelial cell types. Am J Anat 141:537–562
Defries EA, Franks LM (1976) Carcinogenesis in organ culture. In: Balls M, Monneckendam M (eds) Organ culture in biomedial research. Cambridge University Press, Cambridge, p 393
Defries EA, Franks LM (1977) An organ culture method for adult colon from germfree and conventional mice: effects of donor age and carcinogen treatment on epithelial mitotic activity. J Natl Cancer Inst 58:1323–1326
Finney KJ, Ince P, Appleton DR, Sunter JP, Watson AJ (1986) A comparison of crypt-cell proliferation in rat colonic mucosa in vivo and in vitro. J Anat 149:177–188
Finney KJ, Ince P, Appleton DR, Sunter JP, Watson AJ (1987) A trophic effect of epidermal growth factor (EGF) on rat colonic mucosa in organ culture. Cell Tissue Kinet 20:43–56
Hayashi T, Yatani R, Apostol J, Stemmermann GN (1974) Pathogenesis of hyperplastic polyps of the colon: a hypothesis based on ultrastructure and in vitro cell kinetics. Gastroenterology 66:347–356
Johnson HA, Bond VP (1961) A method of labelling tissues with tritiated thymidine in vitro and its use in comparing rates of cell proliferation in duct epithelium, fibroadenoma and carcinoma of the human breast. Cancer 14:639–643
Kovacs L, Potten CS (1973) An estimation of proliferative population size in stomach, jejunum and colon of DBA-2 mice. Cell Tissue Kinet 6:125–134
Lieb LM, Lisco H (1966) In vitro uptake of tritiated thymidine by carcinoma of the human colon. Cancer Res 26:733–740
Lipkin M, Quastler H (1962) Cell population kinetics in the colon of the mouse. J Clin Invest 41:141–146
Loeffler M, Stein R, Wichmann H-E, Potten CS, Kaur P, Chwalinski S (1986) Intestinal cell proliferation. I. A comprehensive model of steady-state proliferation in the crypt. Cell Tissue Kinet 19:627–645
Meinzer HP, Sandblad B (1985) A simulation model for studies of intestine cell dynamics. Comp Meth Prog Biomed 21:89–98
Messier B, Leblond CP (1960) Cell proliferation and migration as revealed by radioautography after injection of thymidineH3 into male rats and mice. Am J Anat 106:247–285
Potten CS, Kovacs L, Hamilton E (1974) Continuous labelling studies on mouse skin and intestine. Cell Tissue Kinet 7:271–283
Pritchett CJ, Senior PV, Sunter JP, Watson AJ, Appleton DR, Wilson RG (1981) Observations on cell proliferation in human colo-rectal tumours in vitro. J Anat 133:119
Reiss B, Williams GM (1978) Induction of DNA repair and suppression of DNA synthesis by carcinogens in colon organ culture. Fed Proc 37:749
Rijke RPC, Plaisier HM, Langendoen NJ (1979) Epithelial cell kinetics in the descending colon of the rat. Virchows Arch [Cell Pathol] 30:85–94
Schiff LJ, Moore SJ (1980) Organ culture of adult rat colonie mucosa on fibrin foam. In vitro 16:893–906
Senior PV, Pritchett CJ, Sunter JP, Appleton DR, Watson AJ (1982) Crypt regeneration in adult human colonie mucosa during prolonged organ culture. J Anat 134:459–469
Senior PV, Sunter JP, Appleton DR, Watson AJ (1984) Morphological studies on the long-term organ culture of colonic mucosa from normal and dimethylhydrazine treated rats. Br J Cancer 49:281–290
Shamsuddin AK, Barrett LA, Autrup H, Harris CC, Trump BF (1978) Long-term organ culture of adult rat colon. Pathol Res Pract 163:362–372
Shorter RG, Spencer RJ, Hallenbeck GA (1966) Kinetic studies of the epithelial cells of the rectal mucosa in normal subjects and patients with ulcerative colitis. Gut 7:593–596
Steel GG, Bensted JPM (1965) In vitro studies of cell proliferation in tumours I. Critical appraisal of methods and theoretical considerations. Eur J Cancer 1:275–279
Sunter JP, Wright NA, Appleton DR (1978) Cell population kinetics in the epithelium of the colon of the male rat. Virchows Arch [Cell Pathol] 26:275–287
Sunter JP, Appleton DR, De Rodriguez MSB, Wright NA, Watson AJ (1979a) A comparison of cell proliferation at different sites within the large bowel of the mouse. J Anat 129:833–842
Sunter JP, Watson AJ, Wright NA, Appleton DR (1979b) Cell proliferation at different sites along the length of the rat colon. Virchows Arch [Cell Pathol] 32:75–87
Tannock IF (1967) A comparison of the relative efficiencies of various metaphase arrest agents. Expl Cell Res 47:345–356
Troughton WD, Trier JS (1969) Paneth and goblet cell renewal in mouse duodenal crypts. J Cell Biol 41:251–268
Tsubouchi S (1981) Kinetic analysis of epithelial cell migration in the mouse descending colon. Am J Anat 161:239–246
Tutton PJM, Barkla DH (1976) Cell proliferation in the descending colon of dimethylhydrazine treated rats and in dimethylhydrazine induced adenocarcinomata. Virchows Arch [Cell Pathol] 21:147–160
Wright NA, Morley AR, Appleton DR (1972) Variation in the duration of mitosis in the crypts of Lieberkühn of the rat; a cytokinetic study using vincristine. Cell Tissue Kinet 5:351–364
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Finney, K.J., Appleton, D.R., Ince, P. et al. Proliferative status of colonic mucosa in organ culture:3H-thymidine-labelling studies and computer modelling. Virchows Archiv B Cell Pathol 56, 397–405 (1988). https://doi.org/10.1007/BF02890043
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DOI: https://doi.org/10.1007/BF02890043