Effects of growth factors during in vitro culture of mouse and rat embryos. E. Yu. Brusentsev, T. N. Igonina, I. N. Rozhkova, D. S. Ragaeva, S. Ya. Amstislavsky ВАВИЛОВСКИЙ

Abstract:

In vitro culture of preimplantation embryos of ICR, HT1AN/Icgn, HT1AC/Icgn and C57BL/6J-Ay mouse strains as well as in OXYS/Icgn rat strain in media containing granulocyte-macrophage colony stimulating factor (GM-CSF) or epidermal growth factor (EGF) has been studied. Both mouse and rat embryos were first frozen in a programmable freezer after a standard protocol using a mixture of glycerol and sucrose as cryoprotectants, thawed and cultured in vitro in R1ECM (rat one-cell embryo culture medium) for 24 hours (mice) and 72 hours (rats). For the in vitro culture experiments with these growth factors, 8-cell frozen-thawed mouse embryos and 2–4-cell frozen-thawed rat embryos were used. Supplementation of the culture medium with GM-CSF improved the rate of embryonic development in HT1AC/Icgn and C57BL/6J-Ay strain mice, while EGF had no effect. The reverse was true of the rats. Supplementation of the culture medium with EGF increased the percentage of developing blastocysts in OXYS/Icgn rat strain, while GM-CSF had no effect. Co-culture of four-cell embryos of HT1AN/ Icgn strain mice with more advanced embryonic stages (morulas) of a different strain ICR led to the facilitation preimplantation embryo development. Experimental results presented here reveal the species-specific effects of growth factors on mouse and rat embryos and indicate that co-culture of different stages of embryo development have stimulatory effects on earlier stages.

About The Authors:

E. Yu. Brusentsev. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

T. N. Igonina. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

I. N. Rozhkova. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

D. S. Ragaeva. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

S. Ya. Amstislavsky. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

References:

1. Aflalo E.D., Sod-Moriah U.A., Potashnik G., Har-Vardi I. EGF increases expression and activity of PAs in preimplantation rat embryos and their implantation rate. Reprod. Biol. Endocrinol. 2007;5:4. DOI:10.1186/1477-7827-5-4

2. Amstislavsky S., Brusentsev E., Kizilova E., Igonina T., Abramova T., Rozhkova I. Embryo cryopreservation and in vitro culture of preimplantation embryos in Campbell’s hamster (Phodopus campbelli). Theriogenology. 2015;82:1056-1063. DOI: 10.1016/j.theriogenology.2014.12.013

3. Amstislavsky S.Ya., Igonina T.N., Rozhkova I.N., Brusentsev E.Yu., Rogovaya A.A., Ragaeva D.S., Naprimerov V.A., Litvinova E.A., Plyusnina I.Z., Markel A.L. Rederivation by embryo transfer in strains of laboratory mice and rats. Vavilovskiy zhurnal genetiki i selektsii — Vavilov Journal of Genetics and Breeding. 2013;17(1):147-161.

4. Blake D., Farquhar C. Cleavage stage versus blastocyst stage embryo transfer in assisted reproductive technology. Cochrane Database Syst. Rev. 2007;4:1-61. DOI: 10.1002/14651858.CD002118.pub3

5. Brigstock D.R., Heap R.B., Brown K.D. Polypeptide growth factors in uterine tissues and secretions. J. Reprod. Fertil. 1989;85:747-758. DOI: 10.1530/jrf.0.0850747

6. Brusentsev E.Yu., Igonina T.N., Amstislavsky S.Ya. Traditional and modern approaches to the culturing of preimplantation mammalian embryos in vitro. Ontogenez — Ontogeny. 2014;45(2):73-88.

7. Desai N., Kattal N., AbdelHafez F.F., Szeptycki-Lawson J., Goldfarb J. Granulocyte-macrophage colony stimulating factor (GM-CSF) and co-culture can affect post-thaw development and apoptosis in cryopreserved embryos. J. Assist. Reprod. Genet. 2007;24(6):215-222. DOI: 10.1007/s10815-007-9119-8

8. Ecker D.J., Stein P., Xu Z., Williams C.J., Kopf G.S., Bilker W.B., Abel T., Schultz R.M. Long-term effects of culture of preimplantation mouse embryos on behavior. Proc. Natl Acad. Sci. USA. 2004;101(6):1595-1600. DOI: 10.1073/pnas.0306846101

9. Elaimi A., Gardner K., Kistnareddy K., Harper J. The effect of GMCSF on development and aneuploidy in murine blastocysts. Hum. Reprod. 2012;27(6):1590-1595. DOI: 10.1093/humrep/des108

10. Emiliani S., Van den Bergh M., Vannin A.S., Biramanel J., Englert Y. Comparison of ethylene glycol, 1,2-propanediol and glycerol for cryopreservation of slow-cooled mouse zygotes, 4-cell embryos and blastocysts. Hum. Reprod. 2000;4:905-910. DOI: 10.1093/humrep/15.4.905

11. Hardy K., Spanos S. Growth factor expression and function in the human and mouse preimplantation embryo. J. Endocrinol. 2002;172(2):221-236. DOI: 10.1677/joe.0.1720221

12. Morita Y., Tsutsumi O., Taketani Y. In vitro treatment of embryos with epidermal growth factor improves viability and increases the implantation rate of blastocysts transferred to recipient mice. Am. J. Obstet. Gynecol. 1994;171(2):406-409.

13. Paria B.C., Dey S.K. Preimplantation embryo development in vitro: cooperative interactions among embryos and role of growth factors. Proc. Natl Acad. Sci. USA. 1990;87(12):4756-4760.

14. Renard J.P., Babinet C. High survival of mouse embryos after rapid freezing and thawing inside plastic straws with 1-2 propanediol as cryoprotectant. J. Exp. Zool. 1984;230:443-448. DOI: 10.1002/jez.1402300313

15. Robertson S.A., Sjöblom C., Jasper M.J., Norman R.J., Seamark R.F. Granulocyte-macrophage colony-stimulating factor promotes glucose transport and blastomere viability in murine preimplantation embryos. Biol. Reprod. 2001;64(4):1206-1215. DOI: 10.1095/biolreprod64.4.1206

16. Sjöblom C., Roberts C.T., Wikland M., Robertson S.A. Granulocytemacrophage colony-stimulating factor alleviates adverse consequences of embryo culture on fetal growth trajectory and placental morphogenesis. Endocrinology. 2005;146(5):2142-2153. DOI: 10.1210/en.2004-1260

17. Sjöblom C., Wikland M., Robertson S.A. Granulocyte-macrophage colony-stimulating factor promotes human blastocyst development in vitro. Hum. Reprod. 1999;14(12):3069-3076. DOI:10.1093/humrep/14.12.3069

18. Spindler R.E., Crichton E.G., Agca Y., Loskutoff N., Critser J., Gardner D.K., Wildt D.E. Improved felid embryo development by group culture is maintained with heterospecific companions. Theriogenology. 2006;66(1):82-92. DOI: 10.1016/j.theriogenology.2006.03.021

19. Spindler R.E., Wildt D.E. Quality and age of companion felid embryos modulate enhanced development by group culture. Biol. Reprod. 2002;66(1):167-173. DOI: 10.1095/biolreprod66.1.167

20. Watkins A., Platt D. Mouse embryo culture induces changes in postnatal phenotype including raised systolic blood pressure. PNAS. 2007;104(13):5449-5454. DOI: 10.1073/pnas.0610317104

21. Ziebe S., Loft A., Povlsen B.B., Erb K., Agerholm I., Aasted M., Gabrielsen A., Hnida C., Zobel D.P., Munding B., Bendz S.H., Robertson S.A. A randomized clinical trial to evaluate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) in embryo culture medium for in vitro fertilization. Fertil. Steril. 2013;99(6):1600-1609. DOI: 10.1016/j.fertnstert.2012.12.043