Method for construction of pancreatic cancer organoid
Pancreatic cancer organoids prepared by a method of the present invention sufficiently reflect the interaction between cancer cells and endothelial cells, that is, cross-talk with the vascular niche, and thus they can show the characteristics of cancer-initiating cells (CICs) present in the organism environment, compared to conventional cancer organoids. Accordingly, the clinical applicability and reliability of the screened drug may be further remarkably increased.
1 . A method for preparing pancreatic cancer organoids containing cancer-initiating cells (CICs), comprising steps of:
a) dissociating a biological sample isolated from a pancreatic cancer patient and immobilizing the dissociated biological sample on a growth factor reduced basement membrane matrix;
b) forming pancreatic cancer organoids by culturing the sample immobilized on the growth factor reduced basement membrane matrix in step a);
c) dissociating the pancreatic cancer organoids formed in step b); and
d) co-culturing the dissociated pancreatic cancer organoids with vascular endothelial cells, or adding a vascular endothelial cell conditioned medium to the dissociated pancreatic cancer organoids, followed by culture in an environment excluding growth factors.
2 . The method according to claim 1 , wherein the co-culturing in step d) comprises mixing the pancreatic cancer organoids and the endothelial cells at a ratio of 1:1 to 1:6 and culturing the mixture.
3 . The method according to claim 1 , wherein the dissociating in step a) is performed using collagenase.
4 . The method according to claim 1 , wherein the pancreatic cancer is pancreatic ductal adenocarcinoma.
5 . The method according to claim 1 , wherein step b) of forming the organoids is performed by culturing the dissociated sample of step a) in a DMEM/F12 medium further containing at least one selected from the group consisting of antibiotics, glutamine, B27, N-acetyl-L-cysteine, gastrin, cell signaling inhibitors, and PGE2 (prostaglandin E2).
6 . A method according to claim 1 , further comprising the steps of:
e treating the pancreatic cancer organoid of step d) with a candidate substance; and
f observing cancer cells contained in the pancreatic cancer organoid containing endothelial cells after treatment with the candidate substance.
7 . The method according to claim 6 , wherein the pancreatic cancer is pancreatic ductal adenocarcinoma.