Poster Presentation ESA-SRB-APEG-NZSE 2022

Effect of GM-CSF on bovine in vitro oocyte maturation. (#366)

Annie Whitty 1 , Staci Jennings 1 , Kylie Dunning 1 2 , Karen Kind 3 , Hannah Brown 1 , Jeremy Thompson 1 2 , Mark Nottle 1
  1. The University of Adelaide, School of Biomedicine, & Robinson Research Institute, Adelaide Medical School, The University of Adelaide, Adelaide, SOUTH AUSTRALIA, Australia
  2. Australian Research Council Centre of Excellence for Nanoscale BioPhotonics, Institute for Photonics and Advanced Sensing, Adelaide, South Australia, Australia
  3. School of Animal and Veterinary Sciences, The University of Adelaide, Adelaide, South Australia, Australia

Aims: 

Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) is found in the reproductive tract and has been shown to have beneficial effects when added to embryo culture media in many species including humans [1, 2]. GM-CSF is also found in the ovarian follicle [3] suggesting it may have a role in oocyte maturation. We have previously shown that the addition of GM-CSF to in vitro oocyte maturation (IVM) media improves oocyte developmental competence resulting in increases in blastocyst rate and cell numbers, and implantation and birth rates following frozen embryo transfer in mice.  The present study was undertaken to determine if GM-CSF can improve bovine IVM.  

 

Methods:

Cumulus oocyte complexes (COCs) were aspirated from abattoir derived ovaries and matured in 0 or 10 ng/ml of recombinant bovine GM-CSF (Kingfisher, Assay Matrix) in VitroMat media (ARTLabs Solutions) at 38.5C in 6% CO2, in air. Following fertilisation in Research Vitro Fertilisation media (Cook Medical) in 6% CO2 in air, presumptive zygotes were cultured in Research Vitro Cleave (Cook Medical) and VitroBlast (ARTLabs Solutions in 6% CO2; 7% O2; N2 balance), and embryo development examined.  Day 8 blastocyst cell numbers were also determined by differential staining and fluorescent imaging. Each group contained approximately 40 COCs and the experiment was replicated 10 times. Data was analysed using a paired t-test.

 

Results:

GM-CSF significantly increased day 8 blastocyst rate (81.1% vs 70.5%), blastocyst inner cell numbers (43.7 vs 38.3 ), total cell numbers (141.6 vs 128.1), and ICM:TE (0.50 vs 0.44) ratio compared with the control group .

 

Conclusion: 

In conclusion our results suggest that adding GM-CSF to bovine IVM media, improves bovine embryo development.

  1. Granulocyte–macrophage colony-stimulating factor promotes human blastocyst development in vitro. C. Sjöblom, M. Wikland, S.A. Robertson ; Human Reproduction, Volume 14, Issue 12, 1 December 1999, (pg. 3069–3076)
  2. Robertson SA, Sjoblom C, Jasper MJ, Norman RJ, Seamark RF. Granulocyte-macrophage colony-stimulating factor promotes glucose transport and blastomere viability in murine preimplantation embryos, Biology of Reproduction, 2001, vol. 64 (pg. 1206-1215)
  3. Ovarian immune cells express granulocyte-macrophage colony-stimulating factor (GM-CSF) during follicular growth and luteinization in gonadotropin-primed immature rodents. K Tamura 1, H Tamura, K Kumasaka, A Miyajima, T Suga, H Kogo. Mol Cell Endocrinol. 1998 Jul 25;142(1-2): (pg.153-63).