Organoids Construction and Characterization
I. Organoid construction process:
Second, the key factors for successful organoid construction:
· Accurate selection of tumor lesions to ensure the amount of tumor cells, freshness and cleanliness of the sample
· Accurate selection of digestion reagents and precise control of digestion time to prevent excessive digestion.
· Collect as many single cells and small cell clusters as possible after digestion and remove incompletely digested tissue.
· Appropriate cell implantation density and high percentage of viable cells.
· Reasonable choice of matrix gel concentration and curing method to reduce cell wall sticking after gel planting.
· Appropriate organoid culture reagents to improve the success rate of organoid construction.
· Daily monitoring of organoid growth status and timely treatment to prevent overgrowth of organoids.
III. Organoid identification:
Histomorphology
Source Tissue/Organoid
Colon cancer tumor-like organs and source tumor tissues have high morphological similarity (HE staining)
Immunofluorescence
CEA/CKA/E-cad
Gastric cancer-like organs express the same biomarkers CEA, E-cad, and CK7 as the source tumor tissue