Intra-prostatic Injection of Cancer Cells: A Technique to Deliver Cancer Cells for Establishing Orthotopic Prostate Cancer Mouse Model

Abstract

Source: Anker, J. F. et al. Bioluminescent and Fluorescent Orthotopic Syngeneic Murine Model of Androgen-dependent and Castration-resistant Prostate Cancer. J. Vis. Exp. (2018)

In this video, we demonstrate the procedure for administering intra-prostatic injection of prostate cancer cells into a mouse model. Orthotopic murine models can be used to study the disease origin and response to treatment therapies in a non-invasive and clinically relevant environment.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Pre-operative Mouse Preparation NOTE: House mice in the university animal facility for at least one week before surgery to allow for adequate adaptation and minimize animal stress. Steps 1.1-1.6 are performed by the surgeon’s assistant. All subsequent surgical steps a…

Representative Results

Figure 1: Anterior prostate lobe, draining lymph nodes, and representative technique for intra-prostatic cell injections. Images of the (A) right anterior prostate lobe (black, *), attached right seminal vesicle (white), right testicle and fat pad (green), and bladder (yellow), (B) bilateral prostate-draining para-aortic lymph nodes (orange), (C)…

Materials

Myc-CaP cell line	ATCC	CRL-3255	Verify as mycoplasma-free before use
Micro-dissecting scissors	Roboz	RS-5910
Graege forceps	Roboz	RS-5135
Graefe tissue forceps	Roboz	RS-5150
Olsen-Hegar needle holder with suture cutters	FST	12002-12
50 µL Syringe 705 RN SYR	Hamilton	7637-01	Sterilize by ethylene oxide gas
28-gauge Needles Small Hub RN H	Hamilton	7803-02	Point style 4, Angle 45°, Length 0.75 in. Sterilized by ethylene oxide gas
RPMI	Gibco	18875-093
FBS	Gibco	10437-028
Pencillin/Streptomycin	Gibco	15140-122
PBS	Gibco	14190-144
Matrigel basement membrane matrix, phenol red-free, LDEV-free	Corning	356237	Thaw on ice in 4°C overnight before use
Isoflurane	Henry Schein	11695-0500-2	Acquired from Northwestern University Center for Comparative Medicine (CCM)
Buprenorphine hyrochloride 0.3 mg/mL		12496-0757-5	Controlled substance, acquired from Northwestern University CCM
26 5/8-gauge syringe	BD	309597	For meloxicam and buprenorphine injections
Ophthalmic ointment lubrication	Akron	17478-162-35
Betadine surgical scrub (povidoneiodine, 7.5%)	Purdue Products	67618-151-16
Sterile non-adhering pads	Moore Medical	10775
Sterile alcohol wipes	Fisher Scientific	22-363-750
Surgical microscope Stemi DV4	Zeiss
Sterile polyester tipped applicators	Puritan	25-806 1PD
5-0 vicryl absorbable reverse cutting needle sutures	eSutures	J493G
4-0 nylon monfilament nonabsorbable reverse cutting needle sutures	eSutures	699H
Glass bead sterilizer	Fine Science Tools	18000-45
Sterile saline 0.9% sodium chloride	Hospira	0409-4888-02