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

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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).

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