Prostate Organoid Assay: A Matrix Gel Ring-based Ex Vivo Culture Technique to Study the Differentiation Capacity of Prostate Epithelial Cells

Published: April 30, 2023

Abstract

Source: Crowell, P. D. et al. Evaluating the Differentiation Capacity of Mouse Prostate Epithelial Cells Using Organoid Culture. J. Vis. Exp. (2019)

In this video, we demonstrate the culturing of isolated mouse prostate epithelial cells using a matrix gel-ring based culture technique. This approach can provide complementary information about the differentiation capacity of prostate epithelial cells to form 3D glandular structures in response to genetic or pharmacological manipulation.

Protocol

1. Plating Sorted Prostate Epithelial Cells into Primary Mouse Organoid Culture —  TIMING: 2-3 H (Excluding Poly-HEMA-coated Plate Preparation) NOTE: Plates are coated with Poly-HEMA to prevent 2D colony formation on the surface of the well beneath the matrix gel. Prepare Poly-HEMA-coated plates 1 day prior to plating sorted basal or luminal prostate epithelial cells into mouse organoid culture. Thaw 1 mL aliquots of reduced growth factor matrix …

Açıklamalar

The authors have nothing to disclose.

Materials

Fetal Bovine Serum (FBS)  Sigma  F8667
B-27 Supplement (50x), Serum Free  Thermo Fisher Scientific  17504044
Advanced DMEM/F-12  Thermo Fisher Scientific  12634010
µ-Dish 35 mm, high   Ibidi 81156
Matrigel GFR Membrane Matrix  Corning CB-40230C
Penicillin-Streptomycin (10,000 U/ mL)  Thermo Fisher Scientific  15-140-122
Poly(2-hydroxyethyl methacrylate) (Poly-HEMA)  Sigma  P3932-25G
Y-27632 dihydrochloride (ROCK inhibitor)  Selleck Chemical  S1049-50MG
A83-01  Tocris  2939
(DiHydro)testosterone (5αAndrostan-17β-ol-3-one)  Sigma  A-8380
GlutaMAX Thermo Fisher Scientific  35050061
Normocin  Thermo Fisher Scientific  ant-nr-1
Recombinant Human EGF, Animal Free PeproTech  AF-100-15
Recombinant Human Noggin PeproTech  120-10C
RPMI 1640 Medium, HEPES (cs of 10) Thermo Fisher Scientific 22400105
Halt Phosphatase Inhibitor Thermo Fisher Scientific 78428
Complete Protease Inhibitor Cocktail Sigma  11836145001
Triton X-100 Sigma X100-5ML
Sonic Dismembrator Thermo Fisher Scientific FB120

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Bu Makaleden Alıntı Yapın
Prostate Organoid Assay: A Matrix Gel Ring-based Ex Vivo Culture Technique to Study the Differentiation Capacity of Prostate Epithelial Cells. J. Vis. Exp. (Pending Publication), e20389, doi: (2023).

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