Journal of Assisted Reproduction and Genetics

, Volume 12, Issue 2, pp 78–87

Production of insulin-like growth factor binding proteins (IGFBPS) by human endometrial stromal cells is stimulated by the presence of embryos

  • Hung -Ching Liu
  • Carol Mele
  • Diana Catz
  • Nicole Noyes
  • Zev Rosenwaks
Clinical Assisted Reproduction



To identify IGFBPs among endometrial secretory products and study their role in implantation and early embryo development.


Two-cell CB6F1mouse embryos were cultured alone or with human endometrial stromal cells in RPMI 1640 plus 10% fetal calf serum (FCS) with or without addition of IGF-I (20 Μg/ml), IGF receptor antibody (0.1 Μg/ml), progesterone (P) (20 ng/ml) and relaxin (R) (20 Μg/ml). On the designated day, the medium was changed to protein-free RPMI and incubated for 16 h. Both conditioned medium and conditioned protein-free medium were then collected for protein analysis and immunoradiometric assay. Cells were fixed with 4% paraformaldehyde for immunohistochemical staining.


IGFBP1 (31 kDa), IGFBP2 (36 kDa), IGFBP3 (45 kDa and 50 kDa) and an unknown IGFBP (25 kDa) were identified in conditioned medium of human endometrial stromal cells cultured alone or cocultured with mouse embryos. Secretion of IGFBPs by endometrial stromal cells was stimulated in the presence of mouse embryos as well as by P and R. IGFBP3 appears to be more responsive to embryonic signals. On the other hand, the secretion of IGFBP1 was greatly stimulated by P and R. Immunolocalization revealed that all three BPs were present in both embryonic and endometrial cells and their immunological staining was heavily increased by P and R.


Endometrial stromal cells were able to synthesize and secrete IGFBPs to modify IGF action on embryo development. Secretion of IGFBPs was stimulated by embryonic signals and was hormonally dependent. The fact that IGFBP3 was more responsive to embryonic signals suggests that it may be important in early implantation. On the other hand, IGFBP1 production was highly responsive to both P and R, suggesting that it may be important throughout pregnancy. In addition, the fact that IGFBPs were located in endometrial and embryonic cells may suggest that these secretory products have autocrine and/or paracrine effects on both types of cells.

Key words

embryo-endometrium coculture IGF embryonic signal IGF binding proteins 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Liu HC, Mele CA, Noyes N, Rosenwaks Z: Endometrial secretory proteins enhance early embryo development. J Assist Reprod Genet 1994;11:217–224PubMedGoogle Scholar
  2. 2.
    De M, Sanford TR, Wood GW: Interleukin-1, interleukin-6 and tumor necrosis factor alpha are produced in the mouse uterus during the estrous cycle and are induced by estrogen and progesterone. Dev Biol 1992;151:297–305PubMedGoogle Scholar
  3. 3.
    Jacobs AL, Carson DD: Uterine epithelial cell secretion of interleukin-1 α induces prostaglandin E2 (PGE2) and PGF{ie86-01} secretion by uterine stromal cells in vitro. Endocrinology 1993;132(1):300–308PubMedGoogle Scholar
  4. 4.
    Jacobs AL, Sehgal PG, Julian J, Carson DD: Secretion and hormonal regulation of interleukin-6 production by mouse uterine stromal and polarized epithelial cells cultured in vitro. Endocrinology 1992;131:1037–1046PubMedGoogle Scholar
  5. 5.
    Robertson SA, Mayrhofer G, Seamark RF: Uterine epithelial cells synthesize granulocyte-macrophage stimulating factor and interleukin-6 in pregnant and nonpregnant mice. Biol Reprod 1992;46:1069–1079PubMedGoogle Scholar
  6. 6.
    Huet-Hudson YM, Chakraborty C, De SK, Suzuki Y, Andrews GK, Dey SK: Estrogen regulates the synthesis of epidermal growth factor in mouse uterine epithelial cells. Mol Endocrinol 1990;4:510–523PubMedGoogle Scholar
  7. 7.
    Tamada H, McMaster MT, Flanders KC, Andrews GK, Dey SK: Cell type-specific expression of transforming growth factor-Β1 in mouse uterus during the periimplantation period. Mol Endocrinol 1990;4:965–972PubMedGoogle Scholar
  8. 8.
    Murphy LJ, Murphy LC, Friesen HG: Estrogen induces insulin-like growth factor expression in the rat uterus. Mol Endocrinol 1988;1:445–450Google Scholar
  9. 9.
    Croze F, Kennedy TG, Schroedter IC, Friesen HC, Murphy LJ: Expression of insulin-like growth factor-1 and insulin-like growth factor-binding protein-1 in the rat uterus during decidualization. Endocrinology 1990;127:1995–2000PubMedGoogle Scholar
  10. 10.
    Bell S, Jackson J, Ashmore J, Zhu H, Tseng L: Regulation of insulin-like growth factor binding protein-1 (hIGFBP-1) synthesis and secretion by progestin and relaxin in long term cultures of human endometrial stromal cells. J Clin Endoc Metab 1991;72:1014–1024Google Scholar
  11. 11.
    Rosenfield RG, Lamson G, Pham H, Oh YM, Conover C, DeLeon D, Donovan SM, Ocrant I, Giudice LC: Insulin-like growth factor binding proteins. Rec Progr Horm Res 1991;16:99–159Google Scholar
  12. 12.
    Knauer DJ, Smith GL: Inhibition of biological activity of multiplication-stimulating activity by binding to its carrier protein. Proc Natl Acad Sci USA 1990;77:7252Google Scholar
  13. 13.
    Rutanen EM, Pekonen F, Makinen T: Soluble 34K binding protein inhibits the binding of insulin-like growth factor-I to its cell receptors in human secretory phase endometrium: evidence for autocrine/paracrine regulation of growth factor action. J Clin Endocrinol Metab 1988;66:173PubMedGoogle Scholar
  14. 14.
    Lai Y, Stein D, Soon Y, Tang Y, Grifo J, Malter H, Talansky B, Cohen J, Liu HC, Rosenwaks Z: Evaluation of vero cell coculture system for mouse embryos in various media. Hum Reprod 1992;7:276–280PubMedGoogle Scholar
  15. 15.
    Hossenlopp P, Seurin D, Segovia-Quinson B, Hardouin S, Binoux M: Analysis of serum insulin-like growth factor binding proteins using western blotting: Use of the method for titration of binding proteins and competitive binding studies. Anal Biochem 1986;154:138–143PubMedGoogle Scholar
  16. 16.
    Daugaday WH, Parker KA, Browsky S, Trivedi B, Kapadia W: Endocrinology 1982;110:575–581PubMedGoogle Scholar
  17. 17.
    Han VKM, D'Ercole AJ, Lund PK: Science (Wash DC) 1987;263:193–197Google Scholar
  18. 18.
    Strech RD, Pinter JE: The embryonic pattern of rat insulinlike growth factor-I gene expression suggests a role in induction and early growth of the liver. Endocrinology 1992;131:2030–2032PubMedGoogle Scholar
  19. 19.
    Xia P, Tekpetey F, Armstrong D: Effect of IGF-I on pig oocyte maturation, fertilization, and early embryonic development in vitro, and on granulosa and cumulus cell biosynthetic activity. Mol Reprod Dev 1994;38:373–379PubMedGoogle Scholar
  20. 20.
    Harvey MB, Kaye PL: Insulin stimulates protein synthesis in compacted mouse embryos. Endocrinology 1988;116:261–263Google Scholar
  21. 21.
    Harvey MB, Kaye PL: Insulin increases the cell number of the inner cell mass and stimulates morphological development of mouse blastocysts in vitro. Development 1990;110:963–967PubMedGoogle Scholar
  22. 22.
    Giudice LC: Growth factors and growth modulators in human uterine endometrium: their potential relevance to reproductive medicine. Fertil Steril 1994;61:1–17PubMedGoogle Scholar
  23. 23.
    Bell SC, Pat SR, Jackson JA, Waites GT: Major secretory protein of human decidualized endometrium in pregnancy is an insulin-like growth factor binding protein. J Endocrinol 1988;118:317–328PubMedGoogle Scholar
  24. 24.
    Giudice LC, Dsupin BA, Irwin JC: Steroid and peptide regulation of insulin-like growth factor binding proteins secreted by human endometrial stromal cells is dependent on stromal differentiation. JCEM 1992;75:1235–1247PubMedGoogle Scholar
  25. 25.
    Boomsma R, Mavrogianis P, Fazieabas A, Jaffe R, Verhage H: Detection of insulin-like growth factor binding protein-1 in cat implantation sites. Biol Reprod 1994;51:392–399PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1995

Authors and Affiliations

  • Hung -Ching Liu
    • 1
  • Carol Mele
    • 1
  • Diana Catz
    • 1
  • Nicole Noyes
    • 1
  • Zev Rosenwaks
    • 1
  1. 1.The Center for Reproductive Medicine and InfertilityCornell University Medical CollegeNew York

Personalised recommendations