Chemically-defined medium formulation and adaptation method for supporting growth of endothelial cells

Area of Science:

• Cell Biology
• Biotechnology
• Translational Research

Background:

• Chemically-defined (CD) media offer advantages like consistency and transparency for in vitro applications.
• Serum-containing media are common, but protocols for adapting sensitive cells, like endothelial cells, to CD media are lacking.
• Standardized cell culture is crucial for reliable bioassays and drug testing.

Purpose of the Study:

• To develop a streamlined protocol for adapting human umbilical vein endothelial cells (HUVECs) to a custom CD medium.
• To optimize weaning strategies and identify effective surface coatings for serum-free cell culture.
• To implement AI-based image analysis for reproducible monitoring of cell growth during adaptation.

Main Methods:

• Evaluating gradual and stepwise adaptation approaches for HUVECs to CD medium.
• Testing various defined extracellular matrix proteins (fibronectin, laminin, collagen IV) as surface coatings.
• Utilizing a trainable AI-based image analysis tool for real-time confluence assessment.

Main Results:

• Fibronectin significantly enhanced HUVEC attachment and viability during CD medium adaptation compared to laminin and collagen IV.
• The developed protocol successfully adapted HUVECs to CD medium, maintaining cell health across multiple passages.
• AI-based image analysis provided quantifiable and reproducible data on cell growth and confluence.

Conclusions:

• A reproducible, modular framework for adapting HUVECs to CD medium was established, prioritizing cell viability and experimental utility.
• Fibronectin coating is critical for successful serum-free adaptation of HUVECs.
• The methodology supports the development of quantitative, ethically aligned bioassays and advances standardized cell culture practices.