Effects of porcine follicle stimulating hormone, luteinizing hormone and estradiol supplementation in culture medium on ultrastructures of porcine cumulus oocyte complexes (pCOCs)

Abstract

Inhibition of primary oocyte developing to secondary oocyte results from oocyte maturation inhibitor (OMI) which is secreted from oocyte-surrounding cumulus cells (CCs) during forming complexes with an oocyte. Development of primary oocyte occurs when the CCs are dissociated from the oocyte. This research studied the effects of pFSH, LH, and estradiol supplementation in culture medium on ultrastructures of porcine cumulus oocyte complexes (pCOCs) using transmission electron microcopy (TEM) and inverted microscopy. A total of 880 oocytes were isolated from 110 ovaries: an average of 8 oocytes per ovary. The oocytes were round in shape and surrounded by zona pellucida with layers of cumulus cells (CCs), at a diameter ranging between 90 and 150 μm and more than 400 μm. Based on the CCs surrounding an oocyte, pCOCs were classified into 5 types, which were intact-, multi-, partial cumulus cell layers, completely denuded oocyte, and expanded cumulus cell layer, which were found at the percentage of 53.86%, 14.32%, 4.32%, 19.20%, and 8.30%, respectively. The Types I and II pCOCs (intact- and multi-CCs layers) were further cultured at 37 °C with 5% CO 2, 95% air atmosphere, and high humidity for 24–48 h to investigate their morphological changes after hormonal induction. For the pCOCs cultured without hormonal induction, the CCs were still round in shape and remained in contact with an oocyte via a process of granular end point sticking into the zona pellucida. In contrast, for the hormone supplemented groups, morphological alteration of pCOCs were seen after culture of 24–48 h. The CCs shape was changed from round into elongated or columnar in an opposite direction from an oocyte as well as no communication between microvilli of CCs observed. This led into ceasing of OMI secretion. Therefore, changes of CCs morphology were a sign of the beginning of oocyte maturation. Further study is to characterize the granular substance at the end point of CCs that stick into the zona pellucida.