A Two-Step Protocol for Isolation and Maintenance of Lung Cancer Primary 3D Cultures

Area of Science:

• Oncology
• 3D Cell Culture
• Cancer Biology

Background:

• Lung cancer remains a leading cause of cancer mortality, necessitating novel therapeutic strategies.
• Patient-derived organoids (PDOs) are valuable tools for studying lung cancer biology and predicting treatment responses.
• Current challenges in establishing lung cancer PDOs include low success rates and contamination by normal cells.

Purpose of the Study:

• To develop a robust and reliable protocol for establishing and expanding lung cancer organoid cultures.
• To improve the success rate of tumoral PDO establishment and ensure faithful representation of patient tissues.
• To provide guidelines for quality control in lung cancer organoid culture.

Main Methods:

• A two-step protocol was developed for establishing both short-term and long-term 3D lung cancer cultures.
• An optimized medium formulation was created to enhance PDO culture success.
• Rigorous quality control measures were implemented to minimize contamination and ensure culture fidelity.

Main Results:

• Cancer tissue-originated spheroids (CTOSs) demonstrated a 100% success rate and allowed for simultaneous isolation of tumor-infiltrating leukocytes (TILs).
• Patient-derived organoids (PDOs) showed improved establishment success rates with the optimized protocol and quality controls.
• The protocol offers flexibility for different experimental needs and settings.

Conclusions:

• The developed protocol enhances the reliability and success rate of lung cancer organoid culture establishment.
• Robust lung cancer PDO culture protocols are crucial for advancing cancer research and clinical applications.
• This method supports the use of organoids as predictive tools in personalized lung cancer therapy.