Preparing Organoids from Tumors: A 3D In Vitro Tumor Model

Published: April 30, 2023

Abstract

Source: Wadosky, K. M., et al. Generation of Tumor Organoids from Genetically Engineered Mouse Models of Prostate Cancer. J. Vis. Exp. (2019).

Organoids derived from tumors are a type of 3D cell culture model that can maintain the pathological features and organ-specific cell lineage of tumors. In the example protocol, we will see scientists generate organoids from a prostate tumor from a genetically engineered mouse model.

Protocol

Animal procedures described here were performed with the approval of the Institutional Animal Care and Use Committee (IACUC) at the Department of Laboratory Animal Resources, Roswell Park Comprehensive Cancer Center, Buffalo, New York. NOTE: Male mice to be dissected to isolate prostates or prostate tumors for generation of organoids should have at least reached the age of sexual maturity — about 8-10 weeks of age. Specific ages of mice can vary amongst stud…

Representative Results

Figure 1: Flow chart of the protocol for generating prostate tumor organoids. After dissecting the prostate tumor, mince the tissue into 1 mm pieces. Digest the tumor pieces in collagenase, collect the cells, and digest in trypsin to obtain a single cell suspension. After counting cells, resuspend in volume of matrix required for a 1.0 x 106 cell/mL cell concentration. Plate domes in dish …

Materials

0.25% Trypsin+2.21 mM EDTA Sigma 25-053
A83-01 MedChemExpress HY-10432
Advanced DMEM/F12+++ Gibco 12634
Analytical balance Mettler Toledo 30216623
B27 (50x) Gibco 17504044
Collagenase II Gibco 17101015
EHS Sarcoma matrix, Pathclear Lot#19814A10 Manufactured by Trevigen Requistitioned from the National Cancer Institute at the Frederick National Laboratory Holder of grants from the National Cancer Institute can request matrix
HEPES (1 M) Sigma 25-060
human recombinant Epidermal growth factor (EGF) PeproTech AF-100-15
L-glutamine (200 mM) Sigma 25-005
N-Acetyl-L-Cysteine Sigma A9165
Penicillin-Streptomycin Sigma P4333
Precision balance Mettler Toledo 30216561
Single-edge carbon razor blade Fisherbrand 12-640
Y-276632 (Rock Inhibitor) APExBIO A3008

Play Video

Citar este artículo
Preparing Organoids from Tumors: A 3D In Vitro Tumor Model. J. Vis. Exp. (Pending Publication), e20065, doi: (2023).

View Video