Granulosa cells inhibit the resumption of meiosis in bovine oocytes in vitro

Biol Reprod. 1990 Nov;43(5):777-83. doi: 10.1095/biolreprod43.5.777.

Abstract

The oocytes of cattle are not as sensitive as those of laboratory animals to purines, cAMP, or follicular extracts. To study the resumption of meiosis, a method is needed that is capable of inhibiting meiosis completely for a minimum of 24 h. This study was designed to evaluate interrelationships in granulosa-oocyte-cumulus complexes using fresh granulosa cells aspirated from small follicles (1-5 mm) in which the cumulus is normally firmly attached. Selected oocyte-cumulus complexes obtained from a slaughterhouse (n = 2,236) were co-incubated with one of the following: various concentrations of fresh granulosa cells in tissue cultures medium (TCM) 199 or bovine follicular fluid (BFF) either without or after one washing and/or freezing; resuspended granulosa cells previously cultured for 7 days; blood cells; or medium alone. Additionally, oocyte-cumulus complexes were embedded in agar cylinders before incubation with or without cells. The rate of maintenance of intact germinal vesicles (GV) in oocytes after 24 h ranged from 40-77% when 5-100 x 10(6) unwashed cells/ml BFF were used, compared to only 16% in oocytes cultured in BFF alone. The pattern was the same when washed cells were used (30-77%, using 5-100 x 10(6) cells/ml BFF), but they were not as effective as unwashed cells. With TCM-199 and the same five concentrations of cells (5, 10, 25, 50, and 100 x 10(6)/ml), a similar inhibition was obtained with greater than or equal to 25 but not with 5 (3%) or 10 (5%) x 10(6)/ml.(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agar
  • Animals
  • Blood Cells / physiology
  • Cattle
  • Cell Count
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Cells, Cultured
  • Cyclic AMP / pharmacology
  • Dose-Response Relationship, Drug
  • Female
  • Follicular Fluid / physiology
  • Granulosa Cells / cytology
  • Granulosa Cells / drug effects
  • Granulosa Cells / physiology*
  • Meiosis / drug effects
  • Meiosis / physiology
  • Oocytes / cytology
  • Oocytes / drug effects
  • Oocytes / growth & development*
  • Purines / pharmacology

Substances

  • Purines
  • Agar
  • Cyclic AMP