Organoid Research FAQs

Answer: After reviewing a large amount of literature, we found that for constructing organoids of primary tumors and metastatic cancers, the organoid medium suitable for primary tumors is used, such as for the primary and metastatic sites of colorectal cancer, and for breast cancer. Therefore, it is better to use the organoid medium from the primary site.

Answer: The morphology of metastatic organoids is more similar to that of the primary cancer.

Answer: If the organoids are relatively large, between 50-100um, you can add drugs; for smaller organoids, around 50um is sufficient. Generally, 2-3 days after passage, you can add drugs and culture for another 3-4 days before testing.

Answer: Place them on ice for 30 minutes, once solidified, directly place them in 70% ethanol and store overnight at 4°C.

Answer: Tissue can be preserved using a gradient freezing method with cryopreservation solution and stored in liquid nitrogen until conditions are suitable for revival and culture.

Answer: Similar to the method for malignant effusions, centrifuge to recover cells, embed in a matrix gel, and add organoid medium.

Answer: Regardless of the type of contamination, whether bacterial, fungal, or other, the organoids must be discarded. At the same time, pour disinfectants like 84 into the culture dish to prevent spread.

Answer: Flow cytometry is possible, or you can directly pick the organoids you want and expand them. Generally, organoids are visible to the naked eye when they grow to 100um. Under a laminar flow hood, use a 200μL pipette tip to directly pick the organoids and then proceed with passage operations.

Answer: Therefore, generally after plating, drug screening is not performed immediately. Instead, 1-2 media change cycles (3-6 days) are allowed to re-evaluate the condition of each well and select appropriate wells for the experiment.

Answer: Some after 24 hours, some after 48 hours, such as chemotherapy drugs. The time points for each drug are different and should be determined based on the specific drug being studied and by testing time gradients.

Answer: Excluding the impact of culture operation skills, for untreated liver cancer tissue, the success rate of culturing hepatocellular carcinoma organoids is higher compared to treated ones. This may be because the activity of liver cancer cells decreases after treatment, and it is also related to the optimization of the nutrient system.