Hum. Reprod. Advance Access originally published online on February 3, 2005
Human Reproduction 2005 20(5):1349-1358; doi:10.1093/humrep/deh750
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In vitro maturation of human oocytes and cumulus cells using a co-culture three-dimensional collagen gel system
1 Brigham and Women's Hospital, Harvard Medical School, Department of Obstetrics and Gynecology, Boston, MA 02115, 2 University of Massachusetts, Department of Veterinary and Animal Sciences, Amherst, MA 01003 and 3 Tufts University School of Medicine, Department of Anatomy and Cellular Biology, Boston, MA 02111, USA 4 Current address: The University of Kansas Medical Center, Department of Molecular and Integrative Physiology, Kansas City, KS 66160, USA
5 To whom correspondence should be addressed at: Biology Department, McCardell Bicentennial Hall 350, Middlebury College, Middlebury, VT 05753, USA. Email: ccombell{at}middlebury.edu
BACKGROUND: Deficiencies remain in the ability of in vitro-matured human oocytes to acquire full developmental competence and give rise to a healthy pregnancy. A clear deficiency of current systems utilizing human oocytes has been the absence of cumulus cells. In the present study, a three-dimensional (3D) co-culture system exploiting an extracellular matrix was developed and compared to conventional methods for its ability to support maturation of human oocytes. METHODS AND RESULTS: Cumulus cells were embedded into a 3D collagen gel matrix with individual oocytes added to each gel. Oocytes from the same patient cultured in the gel matrix matured to metaphase II at rates similar to those of cumulus-free oocytes cultured in individual microdrops. Following maturation of oocytes and fixation of intact gels, chromatin and cytoskeletal elements were assessed in oocytes and cumulus cells. The activities of the key cell cycle kinases, maturation-promoting factor (MPF) and mitogen-activated protein kinase (MAPK), were compared in oocytes matured under the two culture conditions. Compared with denuded oocytes, co-cultured oocytes exhibited increased MAPK activity, but no difference in MPF levels. CONCLUSIONS: This work characterizes a novel and efficacious culture system that takes advantage of the unique properties of the extracellular matrix, a 3D microenvironment, and the presence of cumulus cells for maturing human oocytes in vitro.
Key words: co-culture/cumulus cells/in vitro maturation/oocyte/three-dimensional collagen gel
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