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 |
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).