Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1975 Aug 1;66(2):316–332. doi: 10.1083/jcb.66.2.316

Occluding junctions and cell behavior in primary cultures of normal and neoplastic mammary gland cells

PMCID: PMC2109558  PMID: 1170179

Abstract

Cells dissociated from normal prelactating mouse mammary glands or from spontaneous mammary adenocarcinomas, when grown at high density on an impermeable substrate, form nonproliferating, confluent, epithelial pavements in which turgid, blister-like domes appear as a result of fluid accumulation beneath the cell layer. To compare the structure of the fluid-segregating cell associations in normal and tumor cell cultures with that of lactating gland in vivo, we have examined such cultures alive and in thick and thin sections and freeze-fracture replicas. Pavement cells in all cases are polarized toward the bulk medium as a lumen equivalent, with microvilli and continuous, well- developed occluding junctions at this surface. Between the pavement and the substrate are other cells, of parenchymal or stromal origin, scattered or in loose piles; these sequestered cells are relatively unpolarized and never possess occluding junctions. Small gap junctions have been found in the pavement layer, and desmosomes may link epithelial cells in any location. Under the culture conditions used, development of the epithelial secretory apparatus is not demonstrable; normal and neoplastic cells do not differ consistently in any property examined. A dome's roof is merely a raised part of the epithelial pavement and does not differ from the latter in either cell or junction structure. We suggest that dome formation demonstrates the persistence of some transport functions and of the capacity to form effective occluding junctions. These basic epithelial properties can survive both neoplastic transformation and transition to culture.

Full Text

The Full Text of this article is available as a PDF (5.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Azarnia R., Michalke W., Loewenstein W. R. Intercellular communication and tissue growth. VI. Failure of exchange of endogeneous molecules between cancer cells with defective junctions and noncancerous cells. J Membr Biol. 1972 Dec 29;10(3):247–258. doi: 10.1007/BF01867858. [DOI] [PubMed] [Google Scholar]
  2. Bennett M. V. Function of electrotonic junctions in embryonic and adult tissues. Fed Proc. 1973 Jan;32(1):65–75. [PubMed] [Google Scholar]
  3. Berry M. N., Friend D. S. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol. 1969 Dec;43(3):506–520. doi: 10.1083/jcb.43.3.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chapman G. S., Jones A. L., Meyer U. A., Bissell D. M. Parenchymal cells from adult rat liver in nonproliferating monolayer culture. II. Ultrastructural studies. J Cell Biol. 1973 Dec;59(3):735–747. doi: 10.1083/jcb.59.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Claude P., Goodenough D. A. Fracture faces of zonulae occludentes from "tight" and "leaky" epithelia. J Cell Biol. 1973 Aug;58(2):390–400. doi: 10.1083/jcb.58.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DEOME K. B., FAULKIN L. J., Jr, BERN H. A., BLAIR P. B. Development of mammary tumors from hyperplastic alveolar nodules transplanted into gland-free mammary fat pads of female C3H mice. Cancer Res. 1959 Jun;19(5):515–520. [PubMed] [Google Scholar]
  7. Das N. K., Hosick H. L., Nandi S. Influence of seeding density on multicellular organization and nuclear events in cultures of normal and neoplastic mouse mammary epithelium. J Natl Cancer Inst. 1974 Mar;52(3):849–861. doi: 10.1093/jnci/52.3.849. [DOI] [PubMed] [Google Scholar]
  8. Decker R. S., Friend D. S. Assembly of gap junctions during amphibian neurulation. J Cell Biol. 1974 Jul;62(1):32–47. doi: 10.1083/jcb.62.1.32. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DiPasquale A., Bell P. B., Jr The upper cell surface: its inability to support active cell movement in culture. J Cell Biol. 1974 Jul;62(1):198–214. doi: 10.1083/jcb.62.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Elsdale T., Bard J. Cellular interactions in morphogenesis of epithelial mesenchymal systems. J Cell Biol. 1974 Oct;63(1):343–349. doi: 10.1083/jcb.63.1.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fayet G., Michel-Béchet M., Lissitzky S. Thyrotrophin-induced aggregation and reorganization into follicles of isolated porcine-thyroid cells in culture. 2. Ultrastructural studies. Eur J Biochem. 1971 Dec 22;24(1):100–111. doi: 10.1111/j.1432-1033.1971.tb19659.x. [DOI] [PubMed] [Google Scholar]
  12. Flaxman B. A., Cavoto F. V. Low-resistance junctions in epithelial outgrowths from normal and cancerous epidermis in vitro. J Cell Biol. 1973 Jul;58(1):219–223. doi: 10.1083/jcb.58.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Flaxman B. A., Van Scott E. J. Growth of normal human mammary gland epithelium in vitro. Cancer Res. 1972 Nov;32(11):2407–2412. [PubMed] [Google Scholar]
  14. Goodenough D. A., Gilula N. B. The splitting of hepatocyte gap junctions and zonulae occludentes with hypertonic disaccharides. J Cell Biol. 1974 Jun;61(3):575–590. doi: 10.1083/jcb.61.3.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Guillouzo A., Oudea P., Le Guilly Y., Oudea M. C., Lenoir P., Bourel M. An ultrastructural study of primary cultures of adult human liver tissue. Exp Mol Pathol. 1972 Feb;16(1):1–15. doi: 10.1016/0014-4800(72)90016-0. [DOI] [PubMed] [Google Scholar]
  16. Heaysman J. E., Pegrum S. M. Early contacts between fibroblasts. An ultrastructural study. Exp Cell Res. 1973 Mar 30;78(1):71–78. doi: 10.1016/0014-4827(73)90039-6. [DOI] [PubMed] [Google Scholar]
  17. Hosick H. L. A note on growth patterns of epithelial tumor cells in primary culture. Cancer Res. 1974 Jan;34(1):259–261. [PubMed] [Google Scholar]
  18. Hosick H. L., Nandi S. Preliminary survey of hormonal influences on multicellular architecture in primary cultures of mammary carcinoma cells. J Natl Cancer Inst. 1974 Mar;52(3):897–902. doi: 10.1093/jnci/52.3.897. [DOI] [PubMed] [Google Scholar]
  19. Hülser D. F., Webb D. J. Relation between ionic coupling and morphology of established cells in culture. Exp Cell Res. 1973 Jul;80(1):210–222. doi: 10.1016/0014-4827(73)90291-7. [DOI] [PubMed] [Google Scholar]
  20. Johnson R. G., Sheridan J. D. Junctions between cancer cells in culture: ultrastructure and permeability. Science. 1971 Nov 12;174(4010):717–719. doi: 10.1126/science.174.4010.717. [DOI] [PubMed] [Google Scholar]
  21. LASFARGUES E. Y. Cultivation and behavior in vitro of the normal mammary epithelium of the adult mouse. Anat Rec. 1957 Jan;127(1):117–129. doi: 10.1002/ar.1091270111. [DOI] [PubMed] [Google Scholar]
  22. Lambiotte M., Vorbrodt A., Benedetti E. L. Expression of differentiation of rat foetal hepatocytes in cellular culture under the action of glucocorticoids: appearance of bile canaliculi. Cell Differ. 1973 Mar;2(1):43–53. doi: 10.1016/0045-6039(73)90005-5. [DOI] [PubMed] [Google Scholar]
  23. Leighton J., Brada Z., Estes L. W., Justh G. Secretory activity and oncogenicity of a cell line (MDCK) derived from canine kidney. Science. 1969 Jan 31;163(3866):472–473. doi: 10.1126/science.163.3866.472. [DOI] [PubMed] [Google Scholar]
  24. Leighton J., Estes L. W., Mansukhani S., Brada Z. A cell line derived from normal dog kidney (MDCK) exhibiting qualities of papillary adenocarcinoma and of renal tubular epithelium. Cancer. 1970 Nov;26(5):1022–1028. doi: 10.1002/1097-0142(197011)26:5<1022::aid-cncr2820260509>3.0.co;2-m. [DOI] [PubMed] [Google Scholar]
  25. Linzell J. L., Peaker M. Intracellular concentrations of sodium, potassium and chloride in the lactating mammary gland and their relation to the secretory mechanism. J Physiol. 1971 Aug;216(3):683–700. doi: 10.1113/jphysiol.1971.sp009547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Linzell J. L., Peaker M. Mechanism of milk secretion. Physiol Rev. 1971 Jul;51(3):564–597. doi: 10.1152/physrev.1971.51.3.564. [DOI] [PubMed] [Google Scholar]
  27. Martinez-Palomo A., Braislovsky C., Bernhard W. Ultrastructural modifications of the cell surface and intercellular contacts of some transformed cell strains. Cancer Res. 1969 Apr;29(4):925–937. [PubMed] [Google Scholar]
  28. Martínez-Palomo A. Ultrastructural modifications of intercellular junctions between tumor cells. In Vitro. 1970 Jul-Aug;6(1):15–20. doi: 10.1007/BF02616130. [DOI] [PubMed] [Google Scholar]
  29. McGrath C. M., Blair P. B. Immunofluorescent localization of mammary tumor virus antigens in mammary tumor cells in culture. Cancer Res. 1970 Jul;30(7):1963–1968. [PubMed] [Google Scholar]
  30. McGrath C. M., Nandi S., Young L. Relationship between organization of mammary tumors and the ability of tumor cells to replicate mammary tumor virus and to recognize growth-inhibitory contact signals in vitro. J Virol. 1972 Feb;9(2):367–376. doi: 10.1128/jvi.9.2.367-376.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. McGrath C. M. Replication of mammary tumor virus in tumor cell cultures: dependence on hormone-induced cellular organization. J Natl Cancer Inst. 1971 Aug;47(2):455–467. [PubMed] [Google Scholar]
  32. Moor H. Freeze-etching. Int Rev Cytol. 1969;25:391–412. doi: 10.1016/s0074-7696(08)60209-0. [DOI] [PubMed] [Google Scholar]
  33. Neupert G. Strukturalterationen bei Leberzellen vährend der Gewebsdissoziation und Monolayerformation. Z Mikrosk Anat Forsch. 1972;86(4):443–464. [PubMed] [Google Scholar]
  34. O'Lague P., Dalen H. Low resistance junctions between normal and between virus transformed fibroblasts in tissue culture. Exp Cell Res. 1974 Jun;86(2):374–382. doi: 10.1016/0014-4827(74)90725-3. [DOI] [PubMed] [Google Scholar]
  35. Orci L., Like A. A., Amherdt M., Blondel B., Kanazawa Y., Marliss E. B., Lambert A. E., Wollheim C. B., Renold A. E. Monolayer cell culture of neonatal rat pancreas: an ultrastructural and biochemical study of functioning endocrine cells. J Ultrastruct Res. 1973 May;43(3):270–297. doi: 10.1016/s0022-5320(73)80039-5. [DOI] [PubMed] [Google Scholar]
  36. Owens R. B., Hackett A. J. Tissue culture studies of mouse mammary tumor cells and associated viruses. J Natl Cancer Inst. 1972 Nov;49(5):1321–1332. [PubMed] [Google Scholar]
  37. Owens R. B., Smith H. S., Hackett A. J. Epithelial cell cultures from normal glandular tissue of mice. J Natl Cancer Inst. 1974 Jul;53(1):261–269. doi: 10.1093/jnci/53.1.261. [DOI] [PubMed] [Google Scholar]
  38. Parks W. P., Scolnick E. M. Murine mammary tumor cell clones with varying degrees of virus expression. Virology. 1973 Sep;55(1):163–173. doi: 10.1016/s0042-6822(73)81018-9. [DOI] [PubMed] [Google Scholar]
  39. Pitelka D. R., Hamamoto S. T., Duafala J. G., Nemanic M. K. Cell contacts in the mouse mammary gland. I. Normal gland in postnatal development and the secretory cycle. J Cell Biol. 1973 Mar;56(3):797–818. doi: 10.1083/jcb.56.3.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Rodewald R., Karnovsky M. J. Porous substructure of the glomerular slit diaphragm in the rat and mouse. J Cell Biol. 1974 Feb;60(2):423–433. doi: 10.1083/jcb.60.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Staehelin L. A. Further observations on the fine structure of freeze-cleaved tight junctions. J Cell Sci. 1973 Nov;13(3):763–786. doi: 10.1242/jcs.13.3.763. [DOI] [PubMed] [Google Scholar]
  42. WEXLER H., MINTON J. P., KETCHAM A. S. A COMPARISON OF SURVIVAL TIME AND EXTENT OF TUMOR METASTASES IN MICE WITH TRANSPLANTED, INDUCED AND SPONTANEOUS TUMORS. Cancer. 1965 Aug;18:985–994. doi: 10.1002/1097-0142(196508)18:8<985::aid-cncr2820180810>3.0.co;2-p. [DOI] [PubMed] [Google Scholar]
  43. Wade J. B., Karnovsky M. J. The structure of the zonula occludens. A single fibril model based on freeze-fracture. J Cell Biol. 1974 Jan;60(1):168–180. doi: 10.1083/jcb.60.1.168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Weiss L., Holyoke E. D. Some effects of hypervitaminosis A on metastasis of spontaneous breast cancer in mice. J Natl Cancer Inst. 1969 Nov;43(5):1045–1054. [PubMed] [Google Scholar]
  45. Yagi M. J. Cultivation and characterization of BALB-cfC3H mammary tumor cell lines. J Natl Cancer Inst. 1973 Dec;51(6):1849–1860. doi: 10.1093/jnci/51.6.1849. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES