Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-12-01T00:08:06.908Z Has data issue: false hasContentIssue false
  • English
  • Français

Human cumulus cell complexes studied in vitro by light microscopy and scanning electron microscopy

Published online by Cambridge University Press:  26 September 2008

Sian Kennedy ,
Marjorie A. England  and
Carla Mills
Sian Kennedy
Affiliation:
Medical School, Leicester University, Leicester, and The Hallam Medical Centre, London, UK.
Marjorie A. England*
Affiliation:
Medical School, Leicester University, Leicester, and The Hallam Medical Centre, London, UK.
Carla Mills
Affiliation:
Medical School, Leicester University, Leicester, and The Hallam Medical Centre, London, UK.
*
Dr Marjorie A. England, Department of Anatomy, Leicester University, University Road, Leicester LE1 7RH, UK. Tel: (0533) 523038. Fax: (0533) 525072.
Get access Rights & Permissions [Opens in a new window]

Summary

Various researchers describe the morphology of cumulus cells (CC) in vitro, but few have investigated their behaviour on plastic. Knowledge concerning the behaviour of human CC could be useful in improving the success of in vitro fertilisation procedures. This study aimed to describe the morphology and behaviour of CC in vitro and to investigate movement on a collagen-coated substrate. Following collection some cumulus were mechanically dissected from those surrounding the oocyte. Cumulus aggregates were cultured over 24 h using Earle's medium supplemented with 8% albumin. Substrata were plastic coverslips coated with collagens I, IV, or mixed collagens. Cumulus cultured over corresponding time periods on uncoated coverslips served as controls. Specimens were fixed and prepared for scanning electron microscopy. Over 24 h the controls began exhibiting the morphological features associated with cell movement: cell surface protrusions changed from blebs to microridges, lamellipodia and leading lamellae; cell shape altered from rounded and upright, to flattened. Extracellularmatrix (ECM) transformed from a thick, sheet-like substance to a thin, fibrous material. By 24 h, cells contacting ECM remained rounded showing few features of movement. Collagens enhanced attachment of CC as a monolayer on the substrate. Cell morphology varied according to the collagen type used. On mixed collagens, cells attached rapidly, appearing to be predominantly non-motile. On collagen type I there was less attachment of cells but increased motility. On collagen type IV there was decreased attachment and the cells remained spherical. In conclusion, collagens enhance the settling of cumulus cells on a plastic substrate and the cells exhibit some specificity in attaching to collagens.

Keywords

Cell movementCollagenCumulus cellsIn vitro fertilisation
Type
Article
Information
Zygote , Volume 2 , Issue 2 , May 1994 , pp. 133 - 147
Copyright
Copyright © Cambridge University Press 1994

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abercrombie, M. (1980). The crawling movement of metazoan cells. The Croonian Lecture 1978. Proc. R. Soc. Lond., Ser. B. 207, 129–47.Google Scholar
Abercrombie, M., Heaysman, J.E.M. & Pegrum, S. (1970 a). The locomation of fibroblasts in culture. I. Movements of the leading edge. Exp. Cell Res. 59, 393–8.CrossRefGoogle Scholar
Abercrombie, M., Heaysman, J.E.M. & Pegrum, S. (1970 b). The locomation of fibroblasts in culture. II. ‘Ruffling’. Exp. Cell Res. 60, 437–44.CrossRefGoogle Scholar
Albercht-Büehler, G. (1976). Filopodia of spreading 3T3 cells: do they have a substrate-exploring function? J. Cell Biol. 69, 275–86.CrossRefGoogle Scholar
Anderson, E. & Albertini, D.F. (1976). Gap junctions between the oocyte and companion follicle cells in the mammalian ovary. J. Cell Biol. 71, 680–6.CrossRefGoogle ScholarPubMed
Armstrong, M.T. & Armstrong, P.B. (1980). The role of the extracellular matrix in the motility of fibroblast aggregates. Cell Motil. 1, 99–112.CrossRefGoogle Scholar
Biggers, J.D., Whittingham, D.G. & Donahue, R.P. (1967). The pattern of energy metabolism in the mouse oocyte and zygote. Proc. Natl. Acad. Sci. USA 58, 560–7.CrossRefGoogle ScholarPubMed
Browne, C.L. & Werner, W. (1984). Intercellular junctions between the follicle cells and oocytes of Xenopus laevis. J. Exp. Zool. 230, 105–13.CrossRefGoogle ScholarPubMed
Buccione, R., Vanderhyden, B., Caron, P. & Eppig, J. (1990). FSH induced expansion of the mouse cumulus oophorus in vitro is dependent on significant factor(s) secreted by the oocyte. Dev. Biol. 138, 16–25.CrossRefGoogle Scholar
Chen, L., Wert, S., Hendrix, M., Russell, P., Cannon, M. & Larsen, W. (1990). Hyaluronic acid synthesis and gap junction endocytosis are necessary for the normal expansion of the cumulus mass. Mol. Reprod. Dev. 26, 236–47.CrossRefGoogle ScholarPubMed
Chen, W.T. (1981). Mechanism of retraction of the trailing edge during fibroblast movement. J. Cell Biol. 90, 187–200.CrossRefGoogle ScholarPubMed
Couchman, J.R., Hook, M., Rees, D.A. & Timpl, R. (1983). Adhension, growth and matrix production by fibroblasts on laminin substrates. J. Cell Biol. 96, 177–83.CrossRefGoogle Scholar
Dekel, N. & Beers, W. (1980). Development of the rat oocyte in vitro: inhibition and induction of maturation in the presence or absence of the cumbulus oophorus. Dev. Biol. 75, 247–54.CrossRefGoogle ScholarPubMed
Dekel, N. & Phillips, D.M. (1979). Maturation of the rat cumulus oophorus: a scanning electron microscopic study. Biol. Reprod. 21, 9–18.CrossRefGoogle ScholarPubMed
Dekel, N., Hillensjo, T. & Kracier, P. (1979). Maturational effects of gonadotrophins on the cumulus-oocyte complex of the rat. Biol. Reprod. 20, 197–7.CrossRefGoogle ScholarPubMed
Ebendal, T. (1974). Scanning electron microscopy of thick embryo nerve fibres and heart fibroblasts on collagen substrata in vitro. Zoom. 2, 99–104.Google Scholar
England, M.A. (1983). The migration of primordial germ cells in avian embryos. In: Current problems in cell Differentiation, McLaren, A. & Wylie, C.C.. Symposium of British Society For Development Biology. Cambridge: Cambridge University Press.Google Scholar
England, M.A., Swan, A.P. & Dale, P. (1986). The migration of amphibian primordial germ cells in the chick embryo. Scanning Electron Microsc. 3, 1175–82.Google Scholar
Eppig, J.J. (1977). Mouse oocyte development in vitro with various culture systems. Dev. Biol. 60, 371–88.CrossRefGoogle ScholarPubMed
Gilula, N.,Epstein, M. & Beers, W. (1978). Cell-to-cell communication and ovulation: a study of the cumulus-oocyte complex. J.Cell Biol. 78, 58–75.CrossRefGoogle ScholarPubMed
Harris, A. (1990). Protrusive activity of the cell surface and the movements of tissue cells. In: Biomechanics and Active Deformation of Cells, ed. Akkas, N.. Berlin: Springer.CrossRefGoogle Scholar
Hartshorne, G.M. (1989). Steroid production by the cumulus: relationship to fertilisation in vitro. Hum. Reprod. 4, 742–5.CrossRefGoogle Scholar
Hay, E.D. (1981). Extracellular matrix. J. Cell Biol. 91, 205–23.CrossRefGoogle ScholarPubMed
Heaysman, J.E.M.. (1978). Contact inhibition of locomotion: a reappraisal. Int. Rev. Cytol. 55, 49–64.CrossRefGoogle ScholarPubMed
Herlands, R. & Schultz, R. (1984). Regulation of mouse oocyte growth: probable nutritional role for intercellular communication between follicle cells and oocytes in oocyte growth. J. Exp. Zool. 229, 317–25.CrossRefGoogle ScholarPubMed
Karnovsky, M.J. (1965). A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J. Cell Biol. 27, 137a.Google Scholar
Kleinman, H.K., Murray, J.C., McGoodwin, E.B. & Martin, G.R. (1978). Connective tissue structure: cell binding to collagen. J. Invest. Dermatol. 7, 9–11.CrossRefGoogle Scholar
Kleinman, H.K., Klebe, F.J. & Martin, G.R. (1981). Role of collagenous matrices in the adhesion and growth of cells. J. Cell Biol. 88, 473–85.CrossRefGoogle ScholarPubMed
Kuwana, T., Miyayama, Y., Kajiwara, Y. & Fujimoto, T. (1987). Behaviour of chick primordial germ cells moving towards gonadal primordium in vitro: scanning electron microscope study. Anat. Rec. 219, 164–70.CrossRefGoogle Scholar
Mahadevan, M.M. & Trounson, A.O. (1985). Removal of the cumulus oophorus from the human oocyte for in vitro fertilisation. Fertil. Steril. 43, 263–7.CrossRefGoogle Scholar
Matsumura, G. & England, M.A. (1993). The isolation of chick primordial germ cells from stages 4–8 embryos. Anat. Rec. 235, 604–10.CrossRefGoogle ScholarPubMed
Nakatsuji, N. & Johnson, K.E. (1982). Cell locomotion in vitro by Xenopus laevis gastrula mesodermal cells. Cell Motil. 2, 149–61.CrossRefGoogle ScholarPubMed
Nottola, S.A., Familiari, G., Micara, G., Aragona, C. & Motta, P.M. (1989). The role of the cumulus-corona cells surrounding in vitro human oocytes and polypronuclear ova: an ultrastructural study. Prog. Clin. Biol. Res. 296, 345–54.Google ScholarPubMed
Oster, G. (1988). Biophysics of the leading lamella. Cell Motil. cytoskel. 10, 164–71.CrossRefGoogle ScholarPubMed
Plumel, M. (1948). Tampon au cacodylate de sodium. Bull. Soc. Chim. Biol. Paris 30, 129–30.Google Scholar
Salustri, A., Yanagishita, M. & Hascall, V. (1990). Mouse oocytes regulate hyaluronic acid synthesis and mucification by FSH-stimulated cumulus cells. Dev. Biol. 138, 26–32.CrossRefGoogle ScholarPubMed
Shutt, D.A. & Lopata, A. (1981). The secretion of hormones during the culture of human preimplantation embryos with corona cells. Fertil. Steril. 36, 413–16.CrossRefGoogle Scholar
Wakely, J. & Lopata, A. (1979). Scanning electron microscopical and histochemical study of the structure and function of basement membranes in the early chick embryo. Proc. R. Soc., Ser. B. 206, 329–52.Google ScholarPubMed