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Updated: Jan 13, 2026

Trophoblast Cell Recovery from Angiogenesis-Tube Formation Assay for Differentiation Marker Expression Analysis
Published on: November 8, 2024
The receptor activator of nuclear factor κB ligand (RANKL) regulates the biological behavior of trophoblasts through
YaLu Fu1, QiuYan Zhao1, YuChan Wang1
1Department of Reproductive Medicine, Affiliated Hospital of Shandong Second Medical University, Weifang, Shandong, 261000, People's Republic of China; School of Clinical Medicine, Shandong Second Medical University, Weifang, Shandong, 261021, People's Republic of China.
Introduction:
Abnormal function of trophoblasts is a primary cause of recurrent spontaneous abortion (RSA). However, the underlying molecular mechanisms remain unclear, and further explanation is urgently needed. This study aims to explore the effect and potential regulatory mechanisms of RANKL in the biological function of trophoblasts.
Methods:
Immunohistochemistry was conducted on villous tissues from RSA patients and people who underwent artificial abortion. Cell proliferation, migration, and invasion were assessed in HTR-8/SVneo and JEG-3 cells using CCK-8, scratch wound healing, and transwell assays. Protein expression was analyzed by western blotting.
Results:
The expression of RANKL was significantly lower in villus of RSA patients compared to healthy controls. Recombinant human RANKL (rhRANKL) promoted the proliferation, migration and invasion of trophoblasts, while rhOPG, α-RANKL and MK-2206 (Akt inhibitor) alleviated the effect of rhRANK. Moreover, it may exert the effect through the PI3K/Akt signaling pathway.
Conclusions:
Our results demonstrated that downregulation of RANKL in villous tissue was associated with RSA, and that it promoted trophoblasts proliferation, invasion, and migration via the PI3K/Akt pathway. This may help elucidate the molecular pathogenesis of impaired embryo adhesion and implantation capacity.
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