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Method Article
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April 30th, 2023
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.
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 are performed by the surgeon using sterile gloves and surgical tools with sterile technique.
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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 an.......
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| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| 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<br/> 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<br/> matrix, phenol red-free, LDEV-free | Corning | 356237 | Thaw on ice in 4°C overnight<br/> before use |
| Isoflurane | Henry Schein | 11695-0500-2 | Acquired from Northwestern University Center for Comparative Medicine (CCM) |
| Buprenorphine hyrochloride 0.3<br/> mg/mL | 12496-0757-5 | Controlled substance, acquired<br/> 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 |
