Growth differentiation factor 9 and cumulus cell supplementation in in vitro maturation culture media enhances the viability of human blastocysts

Abstract

Objective: In vitro maturation (IVM) of immature oocytes can be useful for some infertile patients. In IVM programs, the rates of embryo formation and pregnancy are low. Therefore, it is essential to recognize the main factors involved in regulating oocyte maturation in vitro. The purpose of this study was to investigate the effects of growth differentiation factor 9 (GDF9) and cumulus cell (CC) supplementation in IVM medium on the rates of embryo formation and viability of human blastocysts. Methods: A total of 80 germinal vesicle oocytes from stimulated cycles underwent an IVM program. The oocytes were divided into four groups, where group I consisted of IVM media only and served as the control, group II consisted of IVM+CCs, group III consisted of IVM+GDF9 (200 ng/mL), and group IV consisted of IVM+CCs+GDF9 (200 ng/mL). Intracytoplasmic sperm injection was performed on the IVM oocytes, and the cleavage embryos that were generated were vitrified. Following thawing, the embryos were cultured for 3 additional days, and the viability rates of the developed blastocysts were determined. Results: The maturation rate of the oocytes did not differ significantly across the four groups. The fertilization rate in group II was significantly higher than that in the control group (76.5% vs. 46.2%). Embryo formation was significantly more frequent in all experimental groups than in the control group, while blastocyst formation did not show significant differences in the three experimental groups compared to the control. The mean viability rates in groups II, III, and IV were 58.16%, 55.91%, and 55.95%, respectively, versus 37.78% in the control group (p< 0.05). Conclusion: Supplementation of IVM culture media with GDF9 and CCs enhanced the fertilization, embryo formation, and viability rates of blastocysts generated from vitrified cleavage embryos. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

In vitro maturation of immature oocytes for fertility preservation in cancer patients compared to control patients with fertility problems in an in vitro fertilization program

“…found that cumulus cell supplementation in IVM culture media enhances the viability of human embryos (blastocysts) after IVF. 40 The rates of in vitro matured oocytes in cancer patients and patients with fertility problems in our study were very similar to the published rates of IVM of immature oocytes in the usual IVF program, where 65.0%, 68.7%, 68.9% and 69.7% maturation rates were obtained in different maturation media. 41 , 42 , 43 Oktay et al .…”

“…26 Also Chatroudi et al found that cumulus cell supplementation in IVM culture media enhances the viability of human embryos (blastocysts) after IVF. 40 The rates of in vitro matured Non-significant differences, as revealed by Fisher's exact test oocytes in cancer patients and patients with fertility problems in our study were very similar to the published rates of IVM of immature oocytes in the usual IVF program, where 65.0%, 68.7%, 68.9% and 69.7% maturation rates were obtained in different maturation media. [41][42][43] Oktay et al reported the 45% increase in mature oocyte yield after IVM of immature oocytes after controlled hormonal stimulation of the ovaries in breast cancer patients and a high fertilization rate of these oocytes.…”

Factors Influencing the In Vitro Maturation (IVM) of Human Oocyte

“…During the early development of IVM technology, fetal cord serum (FCS) [ 7 , 55 ], or human follicular fluid (HFF) [ 7 , 55 ], or human peritoneal fluid (HPF) [ 55 ] was added to human oocyte IVM medium. Later, patient’s own serum [ 48 ], or human serum albumin (HSA) [ 40 , 41 , 42 , 56 , 57 ], or synthetic substitutive serum (SSS) [ 9 , 39 ], or other synthetic serum substitutes [ 43 ] were used as a protein source supplement in the maturation medium. Considering the complex mixtures, batch-to-batch variability, and the potential risks of disease/infection transmission with protein supplements from biological sources, HSA and other synthetic serum substitutes are recommended for protein supplementations, as their compositions are relatively pure and well-defined.…”

“…The highest concentration at 0.75 IU/mL did not seem to play any role in enhancing IVM rates [ 20 , 29 , 62 , 63 , 64 ]. In most studies, FSH was usually used in combination with LH [ 43 , 56 , 65 , 66 ]/hCG [ 40 , 41 , 44 , 45 , 46 , 67 ].…”

Human Follicle in vitro Culture Including Activation, Growth, and Maturation: A Review of Research Progress

“…Some other factors also have been proved to promote oocyte maturation or benefit subsequent processes ( Table 3). The addition of GDF9 had no effect on maturation rate, while it significantly enhanced fertilization, embryo formation, and viability rates of blastocysts (82). And the growth hormone (GH) was applied to improve the IVM maturation rate of human oocytes (81).…”