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.,&#…
Disclosures
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)
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).