Mouse Lung Preparation for Flow Cytometry: Generating Cell Suspension from Lung Tissue for Flow Cytometric Analysis

Published: April 30, 2023

Abstract

Source: Moll, H. P., et al. Orthotopic Transplantation of Syngeneic Lung Adenocarcinoma Cells to Study PD-L1 Expression. J. Vis. Exp. (2019).

This video describes the preparation of mouse lung cell suspension for flow cytometric analysis to study PD-L1 expression of lung adenocarcinoma cells.

Protocol

All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Lung Preparation for Flow Cytometry At the desired experimental endpoint, sedate the mouse by a subcutaneous injection of a mixture of ketamine (100 mg/kg of body weight) and xylazine (10 mg/kg of body weight) and euthanize it by cervical dislocation. Soak the carcass in 70% ethanol and secure the mouse on a dissect…

Divulgaciones

The authors have nothing to disclose.

Materials

Mouse lung adenocarcinoma cell line isolated in-house
C57Bl/6 mice F1 of the cross of the two backgrounds may be used (8-12 weeks)
129S mice
RPMI 1640 Medium Life Technologies 11544446
Fetal Calf Serum Life Technologies 11573397
Penicillin/Streptomycin Solution Life Technologies 11548876
L-Glutamine Life Technologies 11539876
Ketasol (100 mg/mL Ketamine) Ogris Pharma 8-00173
Xylasol (20 mg/mL Xylazine) Ogris Pharma 8-00178
Blunt forceps Roboz RS8260
Student Iris Scissors Fine Science Tools 91460-11
DNase I (RNase-Free) New England Biolabs M0303S
Collagenase Type I Life Technologies 17100017
ACK Lysing Buffer Lonza 10-548E

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Citar este artículo
Mouse Lung Preparation for Flow Cytometry: Generating Cell Suspension from Lung Tissue for Flow Cytometric Analysis. J. Vis. Exp. (Pending Publication), e20240, doi: (2023).

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