Senescent Alveolospheres: A Preliminary 3D Model for Exploring Epithelial Senescence and Pro-Fibrotic Signaling

Area of Science:

• Pulmonary Medicine
• Cell Biology
• Biotechnology

Background:

• Idiopathic pulmonary fibrosis (IPF) is a severe lung disease linked to the accumulation of senescent cells.
• Epithelial cell senescence impairs lung regeneration and promotes fibrosis, but studying it in vitro is challenging.
• Physiologically relevant models are needed to understand IPF pathogenesis and explore therapeutic targets.

Purpose of the Study:

• To develop a reproducible 3D alveolosphere model for studying epithelial senescence in IPF.
• To investigate epithelial features and pro-fibrotic signaling in senescent conditions.
• To establish a platform for evaluating senotherapeutic compounds.

Main Methods:

• Cultured alveolar epithelial cells to form alveolospheres with or without basement membrane components.
• Induced cellular senescence using doxorubicin exposure.
• Assessed cell viability, morphology, and expression of epithelial, senescent, pro-fibrotic, inflammatory, and matrix-modulating markers.

Main Results:

• The alveolosphere model demonstrated robust cell viability and maintained key epithelial characteristics.
• Induced senescence increased the expression of senescence-associated, pro-fibrotic, inflammatory, and matrix-modulating markers.
• The model successfully recapitulated features relevant to IPF pathogenesis.

Conclusions:

• A cost-effective and reproducible 3D alveolosphere platform was established for studying epithelial senescence in IPF.
• This model can investigate the association between senescence and pro-fibrotic signaling.
• The platform may facilitate preliminary evaluation of senotherapeutic agents for IPF.