72nd ASRM Scientific Congress & Expo
Salt Lake City, UT, USA
P-678 Wednesday, October 19, 2016
GROWTH FACTORS IGF-I AND IGF-II IN HUMAN EMBRYO CO- CULTURE WITH AUTOLOGOUS GRANULOSA CELL CLUSTERS COMPARED TO REGULAR EMBRYO CULTURE IN IVF.
A. Vithoulkas,a M. J. Levanduski,b V. Goudas,c,a K. Illmensee.a aGen- esis Fertility Center, Patras, Greece; bEmbryology, Westchester Fertility and Reproductive Endocrinology, White Plains, NY; cAdvanced Fertility Center of Texas, Houston, TX.
OBJECTIVE: To investigate and compare the presence of IGF-I and IGF- II in human embryo culture with and without autologous granulosa cell supplementation.
DESIGN: Randomized prospective comparative study.
MATERIALS AND METHODS: 19 IVF couples agreed to participate in this study. For autologous comparison, half of the MII-oocytes from the same patient were assigned randomly either to co-culture (group I) or to regular culture (group II), treated for ICSI and cultured in microdrops of Irvine Continuous Single Culture Medium with 15% SSS. As control, autologous granulosa cell clusters were cultured alone (group C). On day 3, 50ml of su- pernatants from group I, II and control group C were collected and stored frozen. Supernatants were analyzed for IGF-I and IGF-II with growth factor-specific magnetic bead panels by Lab Supplies Inc., Athens, Greece.
RESULTS: IGF-I was not detectable in group II, although it was detected in considerable levels in group I and in the control group C. The difference in concentration of IGF-I between groups I and C was not statistically significant (table 1). IGF-II was detected in all groups (see Table 1). However, IGF-II concentration in group II was at significantly higher levels when compared to group I and group C (p < 0.05 and < 0.04 respectively, two- tailed unpaired t-tests).
CONCLUSIONS: We identified IGF-I both in co-culture and control supernatants but not in regular IVF culture, confirming that IGF-I is secreted from granulosa cells. We now propose that IGF-I is another essential growth factor for embryo culture. Investigators have in the past shown an anti- apoptotic effect of IGF-I on cell growth and morphology when added separately in embryo culture media without co-culture. Separately, we noted that IGF-II is present in samples from all groups, but excessively higher in samples from regular IVF culture, thus suggesting that granulosa cells regulate the level of IGF-II in the co-culture. We have previously documented that granulosa-cell supplementation to human embryo culture improves embryo development, due to beneficial contribution of growth factors (Ref. 1). Sup- plementation of embryo culture with granulosa cell clusters is therefore recommended for IVF.
1. A. Vithoulkas, M. Levanduski, V.T. Goudas, K. Illmensee Growth factors associated with embryo co-culture and autologous granulosa cell clusters compared to regular embryo culture in IVF. Fertil Steril, 102, 3S, e218, 2014