Gonadal function of the bank vole females depends on the photoperiod. This experiment was to show whether photoperiod applied on the whole animal in vivo would affect the function of ovarian cells in vitro. Granulosa cells from large ovarian follicles of bank vole reared in long or short photoperiod were cultured as monolayers in control or luteinizing hormone supplemented media. Formation of cell colonies, activity of delta5, 3beta-hydroxy steroid dehydrogenase and progesterone secretion were investigated. First colonies of long day cells were formed already on day 1. On day 2 they enlarged and became abundant. Short day cells formed colonies only on day 2. Colonies of similar size to 2 day colonies of long day cells appeared only on day 6. There were also differences in steroid dehydrogenase activity and in progesterone secretion between long and short day control and hormone treated cultures. We conclude that photoperiod applied in vivo affects ovarian cell function in vitro.