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Modulation of mouse sperm-egg interaction, early embryonic development and trophoblastic outgrowth by activated and
S C Juneja1, T L Pfeifer, X M Tang
1Departments of Obstetrics and Gynecology and Pediatrics, University of Florida, 32610, Gainesville, FL, USA.
Abstract:
Exposure of mouse spermatozoa and oocytes duringin vitro fertilization (IVF) to lipopolysaccharide (LPS) and phorbol myristate acetate (PMA) activated macrophages (U937 cell line), but not unactivated macrophages cultureconditioned medium or control medium (RMPI+DMEM with 0.5% FBS) resulted in inhibition of IVF (87.2%), first cleavage (90.8%) and total blastocyst formation 97.5%). The direct coculture of the activated macrophages with 2-cell stage embryos resulted in arrested development (91.2%), an effect that was significantly diminished in the presence of monolayer of human endometrial stromal cells in the coculture (58.3%). In contrast, the majority of 2-cell embryos developed to blastocysts when exposed to unactivated macrophages, or macrophage-stromal cell cocultures (94.1%). The majority of 2-cell embryos cultured in control medium (DMEM/Ham's F12 with 2% FBS) developed to morulae (96.2%), then underwent growth arrest and degeneration. Furthermore, culturing blastocyst stage embryos in the above groups resulted in a significant enhancement of trophoblast outgrowth, particularly in coculture with activated macrophages as compared to any other group (P<0.005). There was a significant increase in the levels of TGF-β, GM-CSF, IL-1α, IL-1β, TNF-α, PGE(2), TXB(2) and LTB(4) released into the culture conditioned medium of activated macrophages compared to unactivated macrophages (P<0.001). These results suggest that the secretory products of activated macrophages, among them those determined in this study, in a stage-specific manner can directly effect sperm-egg interaction, early embryonic development and trophoblastic outgrowth. This data provides further support for the hypothesis that in endometriosis-associated infertility, continuous exposure of spermatozoa, oocytes and early embryos to activated macrophage-derived factors may play a vital role in their survival during transportation and fertilization as well as development during early embryonic stage.
Insights
Activated macrophages significantly inhibit in vitro fertilization (IVF) and early embryo development, but enhance trophoblast outgrowth. Macrophage factors may contribute to endometriosis-associated infertility by affecting gametes and embryos.
Area of Science:
- Reproductive Biology
- Immunology
- Cell Biology
Background:
- Endometriosis is linked to infertility, with activated macrophages implicated.
- Macrophage-derived factors may impact gamete and embryo function.
Purpose of the Study:
- To investigate the effects of activated macrophages on in vitro fertilization (IVF), early embryonic development, and trophoblast outgrowth.
- To identify specific macrophage-derived factors involved in these processes.
Main Methods:
- Exposure of mouse spermatozoa, oocytes, and embryos to activated and unactivated macrophages (U937 cell line) and their conditioned media.
- Coculture of embryos with macrophages and/or human endometrial stromal cells.
- Analysis of IVF rates, cleavage, blastocyst formation, and trophoblast outgrowth.
- Quantification of cytokine and prostaglandin levels in conditioned media.
Main Results:
- Activated macrophages significantly inhibited IVF, first cleavage, and blastocyst formation.
- Direct coculture with activated macrophages arrested embryo development, an effect reduced by stromal cells.
- Unactivated macrophages and macrophage-stromal cell cocultures supported normal embryo development.
- Activated macrophages significantly enhanced trophoblast outgrowth.
- Elevated levels of TGF-β, GM-CSF, IL-1α, IL-1β, TNF-α, PGE(2), TXB(2), and LTB(4) were observed in activated macrophage media.
Conclusions:
- Secretory products of activated macrophages affect sperm-egg interaction, early embryonic development, and trophoblast outgrowth in a stage-specific manner.
- These findings support the hypothesis that macrophage-derived factors contribute to endometriosis-associated infertility by impacting gamete and embryo viability and development.
