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Updated: Jan 3, 2026
In Vitro Assays to Evaluate the Migration, Invasion, and Proliferation of Immortalized Human First-trimester Trophoblast Cell Lines
Published on: March 5, 2019
Macrophage migration inhibitory factor is involved in endovascular trophoblast cell function in vitro
Aleksandra Vilotic1, Milica Jovanovic Krivokuca1, Ivana Stefanoska1
1Laboratory for Biology of Reproduction, Institute for the Application of Nuclear Energy, INEP, University of Belgrade, Banatska 31b, 11080 Belgrade, Serbia.
Abstract:
Macrophage migration inhibitory factor (MIF) is a multifunctional cytokine abundantly present at the feto-maternal interface proposed to play a role in establishment of pregnancy. We have previously shown that pharmacological inhibition of enzymatic activity of MIF decreases extravillous trophoblast invasion and migration in vitro. This study aimed to further elucidate potential role of endogenous trophoblast MIF, and to assess its importance for endovascular trophoblast cell function in particular. Attenuation of MIF by siRNA reduced HTR-8/SVneo cell invasion through Matrigel (59 % of control), expression of integrin α1 (86 % of control) and levels of MMP2 and MMP9 (87 % and 57 % of control, respectively). MIF specific siRNA reduced the ability of HTR-8/SVneo to differentiate in to endothelial-like phenotype, as determined by Matrigel tube formation assay. The total tube length was decreased to 68.6 %, while the number of branching points was reduced to 57.8 % of control. HTR-8/SVneo cell capacity to integrate into HUVEC monolayers was reduced by knock-down of MIF. This could be partly caused by reduced N-cadherin expression to 63 % of control, which decreased with knock-down of MIF, as the expression of this protein was recently shown essential for trophoblast-endothelial interaction. These novel findings indicate a novel role for trophoblast MIF in spiral artery remodeling process.
Insights
Macrophage migration inhibitory factor (MIF) is crucial for pregnancy establishment. Reducing MIF in trophoblast cells impairs invasion, migration, and spiral artery remodeling, highlighting its role in early pregnancy development.
Area of Science:
- Reproductive immunology
- Cell biology
- Biochemistry
Background:
- Macrophage migration inhibitory factor (MIF) is a cytokine found at the feto-maternal interface.
- Previous research indicated MIF inhibition affects trophoblast invasion and migration.
- The role of endogenous trophoblast MIF in pregnancy establishment requires further investigation.
Purpose of the Study:
- To investigate the role of endogenous trophoblast MIF in trophoblast cell function.
- To assess the importance of MIF for endovascular trophoblast cell function.
- To elucidate MIF's role in spiral artery remodeling.
Main Methods:
- Used siRNA to attenuate MIF expression in HTR-8/SVneo cells.
- Assessed cell invasion using Matrigel assays.
- Evaluated differentiation into an endothelial-like phenotype via Matrigel tube formation assays.
- Measured trophoblast cell integration into HUVEC monolayers.
Main Results:
- MIF knockdown significantly reduced HTR-8/SVneo cell invasion, integrin α1 expression, and MMP2/MMP9 levels.
- MIF depletion impaired trophoblast differentiation into an endothelial-like phenotype, reducing tube formation and branching.
- Reduced N-cadherin expression in MIF-knockdown cells diminished their ability to integrate into endothelial monolayers.
Conclusions:
- Trophoblast-derived MIF plays a critical role in regulating trophoblast cell invasion and migration.
- MIF is essential for trophoblast differentiation into an endothelial-like phenotype and interaction with maternal endothelium.
- These findings suggest a novel function for trophoblast MIF in the spiral artery remodeling process during pregnancy establishment.
