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