Conditional Immortalization of Human Cardiac Fibroblasts for Pro-Fibrotic and Anti-Fibrotic Drug Screening

Area of Science:

• Cardiovascular Biology
• Cell Biology
• Pharmacology

Background:

• Heart failure treatment is limited by a lack of suitable human cardiac fibroblast (CFB) models for pre-clinical drug screening.
• Existing models lack phenotypic stability and scalability, hindering anti-fibrotic therapy development.

Purpose of the Study:

• To establish and validate a conditionally immortalized CFB (iCFB) cell line.
• To create a robust platform for screening cardiac fibrosis drugs.

Main Methods:

• Generated a doxycycline-inducible iCFB cell line.
• Evaluated cellular characteristics via molecular, protein, and functional analyses.
• Assessed model utility by exposing it to pro-fibrotic stimuli (TGF-β1, angiotensin II, palmitic acid) and anti-fibrotic compounds (NS8593, pirfenidone).

Main Results:

• The iCFB line's proliferation was controlled by doxycycline; withdrawal induced a quiescent state.
• iCFBs maintained phenotypic fidelity and myofibroblast differentiation capacity up to population doubling 60.
• The model responded to pro-fibrotic stimuli and confirmed the efficacy of NS8593 and pirfenidone.

Conclusions:

• A novel iCFB model was developed with long-term expandability, high biological fidelity, and responsiveness to fibrotic signaling.
• This platform is suitable for mechanistic studies and accelerates anti-fibrotic drug discovery.