Lung-tumor Tissue Explant Culture: A Procedure to Generate and Culture Precision-cut Lung Tumor Slices

Published: April 30, 2023

Abstract

Source: Nagaraj, A. S., et al. Establishment and Analysis of Tumor Slice Explants as a Prerequisite for Diagnostic Testing. J. Vis. Exp. (2018)

Organotypic primary tissue explant cultures, which include precision-cut slices, represent the three-dimensional (3-D) tissue architecture as well as the multicellular interactions of native tissue. This video describes the culturing of murine lung tumor-derived tissue slices using a rotating incubation unit, a system that enables intermittent exposure of tissues to oxygen and nutrients. The protocol describes the methodology to generate and culture precision-cut lung tumor slices for drug response studies.

Protocol

1.Generation of Precision-cut Lung Tumor Slices Transfer the tumor-bearing lungs in HBSS + P/S into a 10 cm tissue culture plate. Separate the lung lobes using sterile scissors and forceps, and select lobes with tumors on the surface for slicing. NOTE: Tumors >3 mm in size are suitable for slicing. Since normal lung tissue surrounding the big tumor compromises the slicing due to differences in tissue stiffness, tumor tissue undergoing slicing should be separated away from the norma…

Representative Results

Figure 1: Schematic representation of the workflow for establishment and analysis of murine NSCLC tumor-derived slice explants. (A) Schematic describing the collection and preparation of tumor-bearing lungs for slicing. Lung lobes are harvested from a mouse and tumor tissue is dissected away from normal tissue. The black arrowhead and asterisk indicate approximately 4 mm and…

Divulgazioni

The authors have nothing to disclose.

Materials

Hank’s Balanced salt solution (HBSS)  Sigma H6648
Penicillin Streptomycin solution Thermo Fischer Scientific  15140-122
Cyanoacrylate adhesive (GLUture) Abbott  32046-90-1
Leica VT1200 S vibrating blade microtome Leica Biosystems 14048142066
Titanium grids Albamma Research and Development MA0036
Slicing blade VWR  PERS60-0138
Slice incubation unit Albamma Research and Development MD2500
6-well plate Sarstedt  83.1839
10 cm tissue culture plate Sarstedt 83.1802

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Citazione di questo articolo
Lung-tumor Tissue Explant Culture: A Procedure to Generate and Culture Precision-cut Lung Tumor Slices. J. Vis. Exp. (Pending Publication), e20254, doi: (2023).

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