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Transcriptomic Profiling Reveals Mechanisms of Anoikis Resistance in Spheroid-Cultured Human Umbilical Cord

Yuma Iwata1,2, Tomofumi Kodama2, Takahiro Ishikawa1,2

  • 1Department of Dentistry and Oral Surgery, Aichi Medical University, 1-1 Yazako-Karimata, Nagakute, 480-1195, Japan.

Tissue Engineering and Regenerative Medicine
|January 20, 2026
PubMed
Abstract

Insights

Spheroid culture enhances mesenchymal stem cell survival by resisting anoikis, a cell death pathway. This is mediated by PI3K/Akt and hypoxia signaling, improving stem cell therapy potential.

Area of Science:

  • Cell Biology
  • Regenerative Medicine
  • Stem Cell Biology

Background:

  • Mesenchymal stem cells (MSCs) show therapeutic promise but suffer poor survival post-transplantation due to anoikis.
  • Anoikis, triggered by cell detachment, limits the efficacy of MSC-based therapies.
  • Spheroid culture may improve MSC survival, but the underlying mechanisms against anoikis are not fully understood.

Purpose of the Study:

  • To investigate the mechanisms by which spheroid culture confers anoikis resistance in human umbilical cord-derived MSCs (UC-MSCs).
  • To identify key signaling pathways involved in spheroid-mediated anoikis resistance.

Main Methods:

  • Human UC-MSCs were cultured as spheroids and monolayers.
  • RNA sequencing compared gene expression profiles between spheroid and monolayer cultures.
  • Functional assays utilized PI3K/Akt and HIF-1 pathway inhibitors to assess their role in anoikis resistance.

Main Results:

  • Spheroid UC-MSCs displayed significantly enhanced anoikis resistance compared to monolayer cultures.
  • Transcriptomic analysis revealed a regulated apoptotic threshold with downregulated executioner genes.
  • Activation of PI3K/Akt and HIF-1α pathways was identified as crucial for anoikis resistance.

Conclusions:

  • Spheroid formation induces a survival-promoting gene expression profile in UC-MSCs.
  • PI3K/Akt and hypoxia signaling pathways are key mediators of spheroid-enhanced anoikis resistance.
  • Understanding these mechanisms can guide strategies to improve stem cell therapy outcomes.
Keywords:
AnoikisApoptosisMesenchymal stem cellsSignaling pathwaySpheroidsUmbilical cord

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