The influence of human or ovine leptin on in vitro culture of preantral follicles (PFs) isolated from sheep ovaries was investigated. Among the 12 different concentrations (0-1000 ng/mL) of human leptin tested, proportion of PFs exhibiting growth, mean increase in diameter, antrum formation, and maturation of the oocytes to Mil stage were the best in 10 ng/mL. Culture of sheep ovarian PFs in TCM 199 supplemented with 10 ng/mL of human or ovine leptin FSH (2.5 mu g/mL), thyroxine (1 mu g/mL), insulinlike growth factor I (10 ng/mL), and GH (1 mIU/mL) resulted in significantly (P <= 0.05) greater average increase in diameter (11 and 9 vs. 6 1.im), better proportions of PFs exhibiting growth (66% and 58% vs. 48%), antrum formation (51% and 51% vs. 34%), and maturation of oocytes to MIL stage (24% and 22% vs. 7%) than the control medium. It is concluded that (1) the optimum dose of leptin for the growth of sheep PFs in vitro was 10 ng/mL, (2) human or ovine leptin supported similar development in vitro of PFs in sheep, (3) inclusion of leptin along with FSH, thyroxine, insulinlike growth factor I, and GH resulted in only a marginal further improvements in in vitro development of sheep PFs'. (C) 2016 Elsevier Inc. All rights reserved.