Skip to main content

Advertisement

Log in

Exogenous growth factors do not affect the development of individually cultured murine embryos

  • Embryo Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

The objective of this study was to evaluate the effects of multiple growth factors on the development of individually cultured murine embryos.

Methods

Embryos produced by in vitro fertilization using in vitro (IVM) or in vivo (IVO) matured oocytes from three strains of mice (CF1, Swiss Webster, B6D2F1) were cultured individually (10 μl) in the absence (control) or presence of growth factors (paf, epidermal growth factor [EGF], insulin-like growth factor 1 [IGF-1], and granulocyte-macrophage colony-stimulating factor [GM-CSF]). Blastocyst formation, hatching, and blastocyst cell numbers (trophectoderm, inner cell mass, and total) were evaluated on days 4 and 5 of culture. Post-hatching development of CF1 IVO embryos was also evaluated in vitro and in vivo.

Results

The presence of growth factors did not improve the proportion of embryos forming blastocysts or initiating hatching for any of the types of embryos tested. The only significant (P < 0.05) effect of growth factors was a decrease in the proportion of embryos that formed blastocysts by day 5 in CF1 IVM embryos. The presence of growth factors also did not affect blastocyst cell numbers. For CF1 IVO embryos, the presence of growth factors during culture did not affect the proportion of embryos that attached to fibronectin-coated dishes, the size of the resulting outgrowths, or in vivo development following transfer.

Conclusion

Combinations of paf, EGF, GM-CSF, and IGF-1 did not improve development of murine embryos cultured individually in a sequential medium containing a defined protein source.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Cortezzi SS, Cabral EC, Trevisan MG, Ferreira CR, Setti AS, DPdAF B, et al. Prediction of embryo implantation potential by mass spectrometry fingerprinting of the culture medium. Reproduction. 2013;145:453–62.

    Article  CAS  PubMed  Google Scholar 

  2. Krisher RL, Heuberger A, Paczkowski M, Stevens J, Popsil C, Prather RS, et al. Applying metabolomic analyses to the practice of embryology: physiology, development and ART. Reprod Fertil Dev. 2015;27:602–20.

    Article  CAS  PubMed  Google Scholar 

  3. Katz-Jaffe MG, McReynolds S, Gardner DK, Schoolcraft WB. The role of proteomics in defining the human embryonic secretome. Mol Hum Reprod. 2009;15:271–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Dominguez F, Gadea B, Mercader A, Esteban FJ, Pellicer A, Simón C. Embryologic outcome and secretome profile of implanted blastocysts obtained after coculture in human endometrial epithelial cells verus the sequential system. Fertil Steril. 2010;93:774–82.

    Article  CAS  PubMed  Google Scholar 

  5. Rubio I, Galán A, Larreategui Z, Ayerdi F, Bellver J, Herrero J, et al. Clinical validation of embryo culture and selection by morphokinetic analysis: a randomized, controlled trial of the EmbryoScope. Fertil Steril. 2014;102:1287–94.

    Article  PubMed  Google Scholar 

  6. Krisher RL, Schoolcraft WB, Katz-Jaffe MG. Omics as a window to view embryo viability. Fertil Steril. 2015;103:333–41.

    Article  PubMed  Google Scholar 

  7. Gardner DK, Meseguer M, Rubio C, Treff NR. Diagnosis of human preimplantation embryo viability. Hum Reprod Update. 2015;21:727–47.

    Article  CAS  PubMed  Google Scholar 

  8. Lane M, Gardner DK. Effect of incubation volume and embryo density on the development and viability of mouse embryos in vitro. Hum Reprod. 1992;7:558–62.

    Article  CAS  PubMed  Google Scholar 

  9. Ebner T, Shebl O, Moser M, Mayer RB, Arzt W, Tews G. Group culture of human zygotes is superior to individual culture in terms of blastulation, implantation and life birth. Reprod BioMed Online. 2010;21:762–8.

    Article  CAS  PubMed  Google Scholar 

  10. Gardner DK, Lane M, Spitzer A, Batt PA. Enhanced raates of cleavage and development for sheep zygotes cultured to the blastocyst stage in vitro in the absence of serum and somatic cells: amino acids, vitamins, and culturing embryos in groups stimulate development. Biol Reprod. 1994;50:390–400.

    Article  CAS  PubMed  Google Scholar 

  11. Wydooghe E, Heras S, Dewulf J, Piepers S, Van den Abbeel E, De Sutter P, et al. Reprod Fertil Dev. 2014;26:717–24.

    Article  CAS  PubMed  Google Scholar 

  12. Spindler RE, Wildt DE. Quality and age of companion felid embryos modulate enhanced development by group culture. Biol Reprod. 2002;66:167–73.

    Article  CAS  PubMed  Google Scholar 

  13. Spindler RE, Crichton EG, Agca Y, Loskutoff N, Critser J, Gardner DK, et al. Improved felid embryo development by group culture is maintained with heterospecific companions. Theriogenology. 2006;66:82–92.

    Article  PubMed  Google Scholar 

  14. Stokes PJ, Abeydeera LA, Leese HJ. Development of porcine embryos in vivo and in vitro; evidence for embryo ‘cross talk’ in vitro. Dev Biol. 2005;284:62–71.

    Article  CAS  PubMed  Google Scholar 

  15. Gopichandran N, Leese HJ. The effect of paracrine/autocrine interactions on the in vitro culture of bovine preimplantation embryos. Reproduction. 2006;131:269–77.

    Article  CAS  PubMed  Google Scholar 

  16. Kelley RL, Gardner DK. In vitro culture of individual mouse preimplantation embryos: the role of embryo density, microwells, oxygen, timing and conditioned media. Reprod BioMed Online. 2017; In Press; https://doi.org/10.1016/j.rbmo.2017.02.001.

  17. Armant DR. Blastocysts don’t go it alone. Extrinsic signals fine-tune the intrinsic developmental program of trophoblast cells. Dev Biol. 2005;280:260–80.

  18. Singh M, Chaudhry P, Asselin E. Bridging endometrial receptivity and implantation: network of hormones, cytokines, and growth factors. J Endocrinol. 2011;210:5–14.

    Article  CAS  PubMed  Google Scholar 

  19. Maillo V, Sánchez-Calabuig MJ, Loperaa-Vasquez R, Hamdi M, Guiterrez-Adaan A, Lonergan P, et al. Oviductal response to gametes and early embryos in mammals. Reproduction. 2016;152:R127–41.

    Article  CAS  PubMed  Google Scholar 

  20. Thouas GA, Dominguez F, Green MP, Viella F, Simon C, Gardner DK. Soluble ligands and their receptors in human embryo development and implantation. Endocr Rev. 2015;36:92–130.

    Article  CAS  PubMed  Google Scholar 

  21. O’Neill C. The potential roles for embryotrophic ligands in preimplantation embryo development. Hum Reprod Update. 2008;14:275–88.

  22. National Research Council (U.S.A.). Committee for the update of the guide for the care and use of laboratory animals. Guide for the care and use of laboratory animals. 8th ed. Washington D.C.: National Academies Press; 2011.

    Google Scholar 

  23. Herrick JR, Strauss KJ, Schneiderman A, Rawlins M, Stevens J, Schoolcraft WB, et al. The beneficial effects of reduced magnesium during the oocyte-to-embryo transition are conserved in mice, domestic cats, and humans. Reprod Fertil Dev. 2015;27:323–31.

    Article  CAS  PubMed  Google Scholar 

  24. Herrick JR, Paik T, Strauss KJ, Schoolcraft WB, Krisher RL. Building a better mouse embryo assay: effects of mouse strain and in vitro maturation on sensitivity to contaminants of the culture environment. J Assist Reprod Genet. 2016;33:237–45.

    Article  PubMed  Google Scholar 

  25. Bakhtari A, Ross PJ. DPPA3 prevents cytosine hydroxymethylation of the maternal pronucleus and is required for normal development in bovine embryos. Epigenetics. 2014;9:1271–9.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Steele KH, Hester JM, Stone BJ, Carrico KM, Spear BT, Fath-Goodin A. Nonsurgical embryo transfer device compared with wurgery for embryo transfer in mice. J Am Assoc Lab Anim Sci. 2013;52:17–21.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Stone BJ, Steele KH, Fath-Goodin A. A rapid and effective nonsurgical artificial insemination protocol using the NSET™ device for sperm transfer in mice without anesthesia. Transgenic Res. 2015;24:775–81.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. O’Neill. Autocrine mediators are required to act on the embryo by the 2-cell stage to promote normal development and survival of mouse preimplantation embryos in vitro. Biol Reprod. 1998;58:1303–9.

    Article  PubMed  Google Scholar 

  29. Dardik A, Schultz RM. Blastocoel expansion in the preimplantation mouse embryo: stimulatory effect of TGF-α and EGF. Development. 1991;113:919–30.

    CAS  PubMed  Google Scholar 

  30. Desai N, Lawson J, Goldfarb J. Assessment of growth factor effects on post-thaw development of cryopreserved mouse morulae to the blastocyst stage. Hum Reprod. 2000;15:410–8.

    Article  CAS  PubMed  Google Scholar 

  31. O’Neill C. Evidence for the requirement of autocrine growth factors for development of mouse preimplantation embryos in vitro. Biol Reprod. 1997;56:229–37.

  32. Spanos S, Becker DL, Winston RML, Hardy K. Anti-apoptotic action of insulin-like growth factor-I during human preimplantation embryo development. Biol Reprod. 2000;63:1413–20.

    Article  CAS  PubMed  Google Scholar 

  33. Chin PY, Macpherson AM, Thompson JG, Lane M, Robertson SA. Stress response genes are suppressed in mouse preimplantation embryos by granulocyte-macrophage colony-stimulating factor (GM-CSF). Hum Reprod. 2009;24:2997–3009.

    Article  CAS  PubMed  Google Scholar 

  34. Sjöblom C, Wikland M, Robertson SA. Granulocyte-macrophage colony-stimulating factor promotes human blastocyst development in vitro. Hum Reprod. 1999;14:3069–76.

    Article  PubMed  Google Scholar 

  35. Robertson SA, Sjöblom C, Jasper MJ, Norman RJ, Seamark RF. Granulocyte-macrophage colony-stimulating factor promotes glucose transport and blastomere viability in murine preimplantation embryos. Biol Reprod. 2001;64:1206–15.

    Article  CAS  PubMed  Google Scholar 

  36. Wood SA, Kaye PL. Effects of epidermal growth factor on preimplantation mouse embryos. J Reprod Fertil. 1989;85:575–82.

    Article  CAS  PubMed  Google Scholar 

  37. Radonjic-Lazovic G, Roudebush WE. The effect of short- vs. long-term platelet-activating factor exposure on mouse preimplantation embryo development. Early Pregnancy. 1995;1:196–200.

    CAS  PubMed  Google Scholar 

  38. de Moraes AAS, Hansen PJ. Granulocyte-macrophage colony-stimulating factor promotes development of in vitro produced bovine embryos. Biol Reprod. 1997;57:1060–5.

    Article  PubMed  Google Scholar 

  39. Chi MMY, Schlein AL, Moley KH. High insulin-like growth factor 1 (IGF-1) and insulin concentrations trigger apotosis in the mouse blastocyst via down-regulation of the IGF-1 receptor. Endocrinology. 2000;141:4784–92.

    Article  CAS  PubMed  Google Scholar 

  40. Jousan FD, Hansen PJ. Insulin-like growth factor-I as a survival factor for the bovine preimplantation embryo exposed to heat shock. Biol Reprod. 2004;71:1665–70.

    Article  CAS  PubMed  Google Scholar 

  41. Campbell JM, Nottle MB, Vassiliev I, Mithcell M, Lane M. Insulin increases epiblast cell number of in vitro cultured mouse embryos via the PI3K/GSK3/p53 pathway. Stem Cells Dev. 2012;21:2430–41.

    Article  CAS  PubMed  Google Scholar 

  42. Jin XL, O’Neil C. Systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro. Reprod Biol Endocrinol. 2014;12:35.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Heo JS, Han HJ. PKC and MAPKs pathway mediate EGF-induced stimulation of 2-deoxyglucose uptake in mouse embryonic stem cells. Cell Physiol Biochem. 2006;17:145–58.

    Article  CAS  PubMed  Google Scholar 

  44. Navarrete Santos A, Ramin N, Tonack S, Fischer B. Cell lineage-specific signaling of insulin and insulin-like growth factor I in rabbit blastocysts. Endocrinology. 2008;149:515–24.

    Article  PubMed  Google Scholar 

  45. Jousan FD, Oliveira LJ, Hansen PJ. Short-term culture of in vitro produced bovine preimplantation embryos with insulin-like growth factor-I prevents heat shock induced apoptosis through activation of the phosphatidyl 3-kinase/Akt pathway. Mol Reprod Dev. 2008;75:681–8.

    Article  CAS  PubMed  Google Scholar 

  46. Bonilla AQS, Ozawa M, Hansen PJ. Timing and dependence upon mitogen-activated protein kinase signaling for pro-development actions of insulin-like growth factor 1 on the preimplantation bovine embryo. Growth Hormon IGF Res. 2011;21:107–11.

    Article  CAS  Google Scholar 

  47. Jeong W, Kim J, Bazer FW, Song G. Proliferation-stimulating effect of colony-stimulating factor 2 on porcine trophectoderm cells is mediated by activation of phosphatidylinositol 3-kinase and extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase. PLoS One. 2014;9:e88731.

    Article  PubMed  PubMed Central  Google Scholar 

  48. Chatot CL, Lewis JL, Torres I, Ziomek CA. Development of 1-cell embryos from different strains of moce in CZB medium. Biol Reprod. 1990;42:432–40.

    Article  CAS  PubMed  Google Scholar 

  49. Hadi T, Hammer MA, Algire C, Richards T, Baltz JM. Similar effects of osmolarity, glucose, and phosphate on cleavage past the 2-cell stage in mouse embryos from outbred and F1 hybrid females. Biol Reprod. 2005;72:179–87.

    Article  CAS  PubMed  Google Scholar 

  50. Highet AR, Bianco-Miotto T, Pringle KG, Peura A, Bent S, Zhang J, et al. A novel embryo culture media supplement that improves pregnancy rates in mice. Reproduction. 2017;153:327–40.

    Article  CAS  PubMed  Google Scholar 

  51. Kawamura K, Chen Y, Shu Y, Cheng Y, Qiao J, Behr B, et al. Promotion of human early embryonic development and blastocyst outgrowth in vitro using autocrine/paracrine growth factors. PLoS One. 2012;7:e49328.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Dobbs KB, Khan FA, Sakatani M, Moss JI, Ozawa M, Ealy AD, Hansen PJ. Regulation of pluripotency of inner cell mass and growth and differentiation of trophectoderm of the bovine embryo by colony stimulating factor 2. Biol Reprod 2013;89:141,1–10.

  53. Ziebe S, Loft A, Povlsen BB, Erb K, Agerholm I, Aasted M, et al. 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;99:1600–9.

    Article  CAS  PubMed  Google Scholar 

  54. Wei Z, Park KW, Day BN, Prather RS. Effect of epidermal growth factor on preimplantation development and its receptor expression in porcine embryoss. Mol Reprod Dev. 2001;60:457–62.

    Article  CAS  PubMed  Google Scholar 

  55. Karagenc L, Lane M, Gardner DK. Granulocyte-macrophage colony-stimulating factor stimulates mouse blastocyst inner cell mass development only when media lack human serum albumin. Reprod BioMed Online. 2005;10:511–8.

    Article  CAS  PubMed  Google Scholar 

  56. Yoon J, Juhn KM, Ko JK, Yoon SH, Ko Y, Lee CY, et al. Effects of oxygen tension and IGF-I on HIF-1α protein expression in mouse blastocysts. J Assist Reprod Genet. 2013;30:99–105.

    Article  PubMed  Google Scholar 

  57. Kurzawa R, Glabowski W, Baczkowski T, Wiszniewska B, Marchlewicz M. Growth factors protect in vitro cultured embryos from the consequences of oxidative stress. Zygote. 12:231–40.

  58. Gardner DK. Dissection of culture media for embryos: the most important and less important components and characteristics. Reprod Fertil Dev. 2008;20:9–18.

    Article  PubMed  Google Scholar 

  59. Biggers JD, Summer MC. Choosing a culture medium: making informed choices. Fertil Steril. 2008;90:473–83.

    Article  PubMed  Google Scholar 

  60. Donjacour A, Liu X, Lin W, Simbulan R, Rinaudo PF. In vitro fertilization affects growth and glucosemetabolism in a sex-specific manner in an outbred mouse model. Biol Reprod 2014;90:80,1-10.

  61. Summers MC, Bhatnagar PR, Lawitts JA, Biggers JD. Fertilization in vitro of mouse ova from inbred and outbred strains: complete preimplantation embryo development in glucose-supplemented KSOM. Biol Reprod. 1995;53:431–7.

    Article  CAS  PubMed  Google Scholar 

  62. Takeuchi M, Seki M, Furukawa E, Takahashi A, Saito K, Kobayashi M, et al. Improvement of implantation potential in mouse blastocysts derived from IVF by combined treatment with prolactin, epidermal growth factor and 4-hydroxyestradiol. Mol Hum Reprod. 2017;23:557–70.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Erik Strait, Caitlyn Graham, and Brittany Winters for excellent care of the mice used for this study.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jason R. Herrick.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Herrick, J.R., Greene-Ermisch, A.F., Schoolcraft, W.B. et al. Exogenous growth factors do not affect the development of individually cultured murine embryos. J Assist Reprod Genet 35, 523–531 (2018). https://doi.org/10.1007/s10815-017-1103-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-017-1103-3

Keywords

Navigation