Effect of human granulosa cells vitrification on the expression of oocyte secreted factors

Abstract

Background: Oocyte vitrification is a helpful fertility preservation technique for women at risk of losing their ovarian functions. Meanwhile, its application extends with the emerging trend of delayed childbearing, postponed marriage, and further diverse ethical, medical, legal, and social implications. Whereas its effect on oocytes has been widely elucidated, the potential impact of this procedure on the biological functions of granulosa cells remains poorly understood. The present study aimed to assess vitrification’s effect on the granulosa cells (GCs). Methods: 35 women with polycystic ovary syndromes who underwent in vitro fertilization (IVF) in Morula IVF Jakarta Clinic were recruited. The expression of prominent oocytes secreted factors (OSFs), including GDF-9 and BMP-15, were measured at messenger RNA (mRNA) levels. GCs from mature and immature oocytes were collected and calculated separately. Relative expression of GDF-9 and BMP-15 was quantified by a real-time quantitative PCR (RT-qPCR). All of the data was analyzed by using SPSS.  Bivariate analysis used the Mann-Whitney test at a 95% confidence level. A significant p-value was ≤ 0.05. Results: The mean age of subjects was 32,39 ± 3,33 years old, with the mean duration of infertility being six years. The mean BMI, anti-Mullerian hormone (AMH) level, and antral follicle count (AFC) were 24.04 kg/m2, 3.43 ng/mL, and 15 follicles, respectively. The basal FSH, LH, estradiol, and progesterone were at the normal level. The expression of GDF-9 decreased significantly (0.5-fold, p < 0.01) in the immature vitrified GC group but was not in the mature-vitrified GC group. Meanwhile, the expression of BMP-15 was stable in all examined groups (P>0.05). Conclusion: Our results suggest that vitrification may alter oocyte maturation, as demonstrated by reduced GDF-9 expression in the immature vitrified GC group.