Cumulus cell features and nuclear chromatin configuration of in vitro matured canine COCs and the influence of in vivo serum progesterone concentrations of ovary donors

Abstract

Phenotype integrity is viewed as an indicator of cumulus-oocyte complex (COC) viability. The objectives of this study were: (a) to observe the influence of cumulus investment expansion on the nuclear chromatin configuration of canine oocytes matured in vitro; (b) to examine the relationship between cumulus cell (CC) expansion and its morphology after in vitro maturation (IVM); (c) to ascertain the influence of in vivo serum progesterone (SP) concentrations of ovary donors on oocyte nuclear maturation, CC phenotypes and degrees of CC expansion of in vitro matured COCs. After 48 h of IVM in modified TCM 199, CCs from grade 1 and 2 COCs were stained with propidium iodide. Oocyte chromatin configuration was visualized by Hoechst 33342 stain. Results showed that oocyte IVM was not influenced by degree of CC expansion (D1, D2 and D3) in COCs. From the CC types (C1, C2 and C3), number of C1 types was higher at D1 expansion and differed from those observed at D2 and D3 expansions. Additionally, rates of apoptosis in D1 CCs were lower than those observed in D2 CCs (p < 0.05). Oocyte nuclear maturation was not influenced by in vivo SP concentrations of ovary donors. On the other hand, D3 expansion prevailed in COCs from bitches at SP > 2.5 ng/ml (p < 0.001). Moreover, in vitro CC apoptosis was associated both with low (0-1 ng/ml) and with high (>5 ng/ml) in vivo SP levels. These findings indicate that morphology of CCs from in vitro matured dog oocytes gives valuable information on viability of COCs and could possibly be used as a parameter in predicting the quality of oocytes destined for in vitro fertilization (IVF) and their outcomes.