Patient-derived Xenograft Modeling: A Technique to Generate Melanoma Mouse Models

Published: April 30, 2023

Abstract

Source: Min Xiao et al. A Melanoma Patient-Derived Xenograft Model. J. Vis. Exp. (2018)

In this video, human-derived melanoma cells are implanted subcutaneously into the flank region of an immunocompromised mouse. The PDX model allows preclinical investigation of melanoma cells that better recapitulate the tumor heterogeneity and melanoma aggressiveness observed in de vivo conditions.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Tumor tissue processing for mouse implantation Surgical excision or surgical biopsy tissue processing Transfer the tissue to a sterile Petri dish and separate the tumor tissue from the surrounding normal tissue as much as possible. Remove necrotic tissue (usually identified as pale-whitish ti…

Representative Results

Figure 1: Alternative implantation methods. (A) Tumors can be processed into either chunks, a slurry suspension, or as a single-cell suspension. (B) All three methods will allow for the growth of tumors in vivo. Shown here are mice subcutaneously implanted with tumor and imaged 12 days after implantation. (C) Shown are tumor growth curves fo…

Divulgaciones

The authors have nothing to disclose.

Materials

1 M Hepes SIGMA-ALDRICH CORPORATION Cat # H0887-100ML
100x PenStrep  Invitrogen Cat # 15140163
1x HBSS-/- (w/o Ca++ or Mg++) MED Cat # MT21-023-CV
2.5% Trypsin  SIGMA-ALDRICH CORPORATION Cat # T4549-100ML 10 mL aliquots stored at –20oC
BSA SIGMA-ALDRICH CORPORATION Cat # A9418-500G
Chlorhexidine Fisher Scientific Cat# 50-118-0313
Collagenase IV (2,000 u/mL) Worthington  Cat #4189 make up in HBSS-/- from Collagenase IV powder stock (Worthington #4189, u/mg indicated on bottle and varies with each lot); freeze 1
DMSO SIGMA-ALDRICH CORPORATION Cat # C6295-50ML
DNase SIGMA-ALDRICH CORPORATION Cat # D4527
EGTA (ethylene glycol bis(2-aminoethyl ether)-N,N,N’N’-tetraacetic acid) Merck Cat # 324626.25
FBS INVITROGEN LIFE TECHNOLOGIES Cat # 16000-044
Fungizone INVITROGEN LIFE TECHNOLOGIES Cat # 15290-018
Gentamicin FISHER SCIENTIFIC Cat # BW17518Z
Isoflurane HENRY SCHEIN ANIMAL HEALTH Cat # 050031
Leibovitz's L15 media  Invitrogen Cat # 21083027
Matrigel Corning Cat # 354230 Artificial extracellular matrix
Meloxicam HENRY SCHEIN ANIMAL HEALTHRequisition # ::Henry Schein Cat # 025115 1-5mg/kg, as painkiller
NOD/SCID/IL2-receptor null (NSG) Mice The Wistar Institute, animal facility Breeding
PVA (polyvinyl alcohol) SIGMA-ALDRICH CORPORATION Cat # P8136-250G
RPMI 1640 Medium (Mod.) 1X with L-Glutamine Fisher Scientific Cat# MT10041CM
Scalpel Feather Cat # 2976-22
Wound clips MikRon Cat #427631

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Citar este artículo
Patient-derived Xenograft Modeling: A Technique to Generate Melanoma Mouse Models. J. Vis. Exp. (Pending Publication), e20349, doi: (2023).

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