Individual Lobe Extraction from Prostate: A Technique to Harvest Individual Prostate Lobes from Mouse Prostate
Abstract
Source: Nath, D. et al. Identification, Histological Characterization, and Dissection of Mouse Prostate Lobes for In Vitro 3D Spheroid Culture Models. J. Vis. Exp. (2018)
This video describes a technique to harvest individual lobes from the prostate gland of a mouse model. Individual lobes can then be used for histological analysis and cancer research.
Protocol
All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Gross Anatomy of the Prostate and Individual Lobe Microdissection
NOTE: This is depicted in Figures 1(i-n) and Figure 2
- Flip the tissue with a pair of blunt forceps so that the dorsal side faces up, showing the dorsal lobes, which resemble a butterfly’s wings.
- Collect the dorsal lobes by holding the lobe with forceps and snipping at the base with scissors (Figure 1j).
- Flip over the remaining tissue to the ventral side.
- Collect the lateral lobes, which are small and usually wrap the urethra on the side, which are wedged between the anterior, ventral, and dorsal lobes (Figure 1k).
- Next, collect the ventral lobes, which are larger than the lateral lobes and lie on the urethra ventrally (Figure 1l).
- Last, harvest the anterior lobes, the largest of the four, by cutting and discarding the urethra (Figure 1m).
- Process the tissue pieces according to experimental needs.
Representative Results
Figure 1: Dissection of the mouse prostate. Step-by-step images for the dissection of the mouse prostate from the UGS: (a) removing fat, (b) removing the urinary bladder, (c) removing the vas deferens, (d) ventral view of the prostate with the urethra, (e) dorsal view of the prostate with the urethra, (f-h) removing the seminal vesicles, (i) ventral view of the prostate, (j) dissecting a dorsal lobe, (k) dissecting a lateral lobe, (l) dissecting a ventral lobe, (m) dissecting an anterior lobe, and (n) the dissected prostate lobes: anterior = AP, dorsal = DP, lateral = LP and ventral =VP.
Figure 2: Anatomy of the mouse prostate lobes. Representative images from 3-month-old mouse UGS after removal of the fat, bladder, and vas deferens. Images show prostate lobes with seminal vesicles and urethra, with dorsal (a and c) and ventral (b and d) views. Top panels (a and b) show untouched images, while bottom panels (c and d) show the lobes outlined in blue (dorsal), red (anterior), yellow (ventral), and green (lateral).
開示
The authors have nothing to disclose.
Materials
| Mouse surgical instruments (Mouse Dissecting kit) | World Precision Instruments | MOUSEKIT | |
| Dissection microscope | |||
| RPMI medium | Thermofisher Scientific | 11875093 | |
| Dissection medium (DMEM + 10%FBS) | Thermofisher Scientific | 11965-084 | |
| Fetal Bovine Serum | Thermofisher Scientific | 10438018 | |
| PBS (Phosphate buffered saline) | Thermofisher Scientific | 10010031 | |
| Collagenase | Thermofisher Scientific | 17018029 | Make 10x stock (10mg/ml) in RPMI, filter sterilize, aliquot and store at -20 °C |
| Syringes and Needles | Fisher Scientific |
