Effect Of Ascorbic Acid On Vitro Growth Of Porcine Oocyte Derived From Pre-Antral Follicle (0,3-0,7 Mm)

Abstract

The ‘optimal’ conditions of the in vitro growth (IVG) process in pre-antral follicles of porcine, since it shared similarities with human is very necessary to increase the supply of grown and matured oocytes for other research. Moreover, as an antioxidant, Ascorbic Acid has shown its function of reducing ROS in culturing of maturation oocytes, but not for pre antral follicles. 2D system in the IVG process of porcine oocytes derived from pre-antral follicles also not have been researched successfully. Thus, this research’s purpose is to examine the role of Ascorbic Acid in in vitro growth of porcine oocytes derived from the pre antral follicles, together evaluate the efficiency of IVG culture of pre-antral follicles among 2D system on normal plastic dish and on extracellular matrix. First, OGCs from pre-antral follicles were culture on IVG for 7-10 days with supplemented of AA (0µM, 50µM, 100µM), then the effect of AA on morphology, Chromatin configuration and Histone modification was examined. Afterward, the ‘optimal’ concentration of AA was used in examined the effect of Laminin and Agarose coated dish for IVG oocyte derived from pre-antral follicles. Based on the final result, AA at a concentration of 50µM maintained the higher quality of oocyte, better cumulus granulosa cells’ proliferation, higher proportion of oocyte survived (71.15%), antral forming (24.77% on Day 3 and 32.18% on Day 7), and higher of GV presence (13%) after the IVG process, compared to 0µM and 100µM. Furthermore, 2D culture system using Laminin and Agarose also presented improved in morphology, survival and antrum formation, duration (up to Day 14 for Agarose coated dish). To sum up, during IVG culture of porcine oocytes derived from pre-antral follicles, AA at a concentration of 50µM and 2D systems available of Laminin and Agarose demonstrate the optimal condition for culturing the OGCs, as it improved morphology, survival, antrum formation and chromosome configuration of oocytes.