Gastric Fundic Glands Isolation: A Method to Isolate Fundic Glands from Human Stomach Tissue
Transcripción
In humans, the fundus is the uppermost part of the stomach. Fundic epithelium consists of numerous gastric glands that secrete gastric juice, which aids in digestion.
To isolate the gastric fundic glands, begin by taking human fundic tissue in a beaker. Add a suitable buffer to the beaker and wash the fundic tissue vigorously. Washing removes the contaminating debris and blood from the fundic tissue.
Transfer the clean tissue to a Petri plate. Using sterile gauze, wipe away the protective mucous layer, thus exposing the underlying epithelial layer. Using forceps, forcefully scrape the uppermost epithelial layer from the tissue.
Transfer the scraped epithelium to another Petri plate and mince it into smaller pieces. Add the minced pieces to a flask containing a buffer supplemented with collagenase enzymes. Incubate at the desired temperature with constant agitation to prevent clumping.
Collagenase digests the extracellular matrix proteins present in the epithelial layer, releasing the dissociated fundic glands into suspension. Filter the suspension into a fresh tube to remove any undissociated clumps.
Keep the glandular filtrate undisturbed on ice to sediment the glands. Discard the supernatant containing cell debris and resuspend the glandular sediment in a fresh buffer. Centrifuge and store the glandular pellet until further use.
