What Are the Differences in Culture Conditions for Different Tumor Organoids?

Tumor organoids are 3D models that recapitulate the characteristics of tumors. The success of their culture lies in customized culture conditions. Given the distinct signaling pathways and stemness requirements across different cancer types, a universal culture medium is not applicable. This Q&A will clarify the key differences.

Q1: Is there a universal culture foundation for tumor organoids?

A: A common framework exists, but no universal medium formula The basic environment is consistent: 37°C incubator with 5% CO₂. Matrigel is mostly used as the matrix. The basal medium is mainly Advanced DMEM/F12, supplemented with antibiotics, HEPES, and N-acetylcysteine to prevent contamination and stabilize the environment. Growth factors and pathway inhibitors are the core variables.

Q2: How to culture colorectal cancer organoids?

A: Dependent on the Wnt pathway, with the most mature formula Key supplements: EGF, Noggin, R-Spondin1, Wnt3a, plus the TGF-β inhibitor A83–01 and nicotinamide. Colorectal epithelial stem cells are highly dependent on Wnt activators, serving as a standard template for digestive tract organoids with a high culture success rate.

Q3: What are the differences between gastric and pancreatic cancer organoids?

A: Both belong to the digestive tract but require additional supplementary factors

• Gastric cancer: Retains core factors for colorectal cancer, with extra supplementation of FGF10 and Gastrin I to enhance the stemness of gastric epithelial cells and maintain glandular structure.

• Pancreatic cancer: Omits Wnt3a, adds FGF10 and HGF, increases R-Spondin1 concentration, inhibits excessive stromal proliferation, and adapts to the pancreatic tumor microenvironment.

Q4: What is special about lung and breast cancer organoids?

A: Independent of the digestive tract system, with highly targeted factors

• Non-small cell lung cancer: Uses Hepes-buffered DMEM as the basal medium, supplemented with EGF, FGF2, Noggin, and a ROCK inhibitor to reduce apoptosis; Wnt activators are not required.

• Breast cancer: Adjusted by subtype. Triple-negative breast cancer requires EGF, FGF2, and heparin. Hormone receptor-positive types exclude Wnt factors and add estrogen analogs to match tumor hormone dependence.

Q5: What are the key precautions to avoid failures in culture?

A: Strictly control sample ex vivo time and ensure precise factor concentrations Solid tumor samples should be processed within 1 hour after isolation to avoid ischemic necrosis. Growth factors should be prepared fresh before use; concentration deviations directly lead to organoid apoptosis or abnormal differentiation.

Founded in 2020, Suzhou XianJueBio Co., Ltd. is an innovative biotechnology enterprise focusing on the R&D, translation, and promotion of new technologies. It has been certified as a National High-Tech Enterprise and a National Science and Technology Small and Medium-sized Enterprise.

Its products cover personalized precision medicine, new drug development, regenerative medicine, and other fields. Relying on three core technology systems — tumor organoids, iPSCs-induced differentiation, and disease-related exosome capture and diagnosis — the company has incubated a variety of medical diagnostic and scientific research products.

It owns multiple technical service platforms including organoid model development, tumor organoid drug sensitivity testing, and scientific research translation/new drug development. With internationally advanced technologies and R&D teams, Suzhou XianJueBio provides organoid culture media (primary, iPSC), extracellular matrix biomimetic hydrogels, cytokines, intelligent working systems for organoid microphysiological models, vascularized microphysiological model chips, exosome diagnostic chips, and other products.