Individual Lobe Extraction from Prostate: A Technique to Harvest Individual Prostate Lobes from Mouse Prostate

프로토콜

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

1. 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.
2. Collect the dorsal lobes by holding the lobe with forceps and snipping at the base with scissors (Figure 1j).
3. Flip over the remaining tissue to the ventral side.
4. 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).
5. Next, collect the ventral lobes, which are larger than the lateral lobes and lie on the urethra ventrally (Figure 1l).
6. Last, harvest the anterior lobes, the largest of the four, by cutting and discarding the urethra (Figure 1m).
7. Process the tissue pieces according to experimental needs.

결과

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-...

자료

Name	Company	Catalog Number	Comments
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