Intracecal Tumor Cell Injection: A Technique to Deliver Cancer Cells for Establishing Orthotopic Colorectal Cancer Mouse Model
Published: April 30, 2023
Abstract
Source: Kochall, S., et al. Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer J. Vis. Exp. (2017)
The video describes the technique for intracecal injection of cancerous cells to develop an orthotopic mouse model with highly uniform tumors. Therefore, the orthotopic mouse model allows realistic simulation of metastasis, providing a valuable tool to study the phenotype changes during tumor growth and dissemination.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Preparation of cell lines for injection NOTE: Use a volume of 20 µL with 100,000 cells for each injection. Use basement membrane matrix (BMM) to prevent leakage and ensure standardized injection. To ensure reproducible results, conduct cell line authentication assays (e.g.,&#…
Divulgaciones
The authors have nothing to disclose.
Materials
| Sevoflurane | AbbVie Germany GmbH & Co. KG | – | |
| Medical oxygen | Air Liquide Medical GmbH | – | |
| Buprenorphine | Temgesic | – | |
| Bepanthen – opthalmic ointment | Bayer Vital GmbH | 10047757 | |
| Normal saline 0.9% (E154) | Serumwerk Bernburg AG | 10013 | |
| Aqua ad injectabilia | Braun | 235144 | |
| 1 mL Syringe (without dead volume) – Injekt-F SOLO | Braun/neoLab | 194291661 | |
| 30G injection needle | BECTON DICKINSON | 304000 | |
| cellulose swabs | Lohmann & Rauscher Deutschland | 13356 | |
| Micro-Adson Forceps | FST – Fine Science Tools | 11018-12 | |
| Iris Scissor – ToughCut | FST – Fine Science Tools | 14058-11 | |
| Olsen-Hegar Needle Holder | FST – Fine Science Tools | 12002-12 | |
| AutoClip Kit | FST – Fine Science Tools | 12020-00 | |
| Table Top Research Anesthesia Machine w/O2 Flush and a Sevoflurane Vaporizer | Parkland Scientific | V3000PS/PK | |
| UltraMicro Pump with Micro4 Controller | World Precision Instruments | UMP3-4 | Equipment for highly controlled orthotopic injection |
| Footswitch for SYS-Micro4 Controller | World Precision Instruments | 15867 | Equipment for highly controlled orthotopic injection |
| Three-axis Manual Micromanipulator | World Precision Instruments | M325 | Equipment for highly controlled orthotopic injection |
| Magnetic Stand for Micromanipulator | World Precision Instruments | M10 | Equipment for highly controlled orthotopic injection |
| Steel Base Plate for M10 Magnetic Stand | World Precision Instruments | 5479 | Equipment for highly controlled orthotopic injection |
| Hot Plate 062 | Labotect | 13854 | |
| Isis – Hair shaver | AESCULAP – Braun | – | |
| Binocular Surgical Microscope | Parkland Scientific | VS-2Z | |
| Matrigel basement membrane matrix (BMM, phenol red free) | CORNING B.V. Life Sciences | 356231 |
Citar este artículo
Intracecal Tumor Cell Injection: A Technique to Deliver Cancer Cells for Establishing Orthotopic Colorectal Cancer Mouse Model. J. Vis. Exp. (Pending Publication), e20341, doi: (2023).