[The establishment of a monolayer culture system of guinea pig chief cells and an enzyme immunoassay system for guinea pig pepsinogen].
For the purpose of studying pepsinogen secretion from gastric chief cells, we established a monolayer culture system of guinea pig chief cells and an enzyme immunoassay (EIA) system specific for guinea pig pepsinogen. Dispersed chief cells were obtained from gastric mucosa of a guinea pig using collagenase, GEDTA, and Percoll solution, suspended in DMEM/F-12 (1/1 containing 10% FCS) media, and cultured for 70hr. Then the monolayer culture system was established. Pepsinogen was purified from gastric mucosa of a guinea pig using DEAE-Sephacel and Sephacryl S-200 columns. Antibody to pepsinogen was raised by immunizing rabbit with the purified pepsinogen. A two-site EIA system was then established using beta-galactosidase-labeled Fab' antibody. The EIA system showed sensitivity to measure above 1.5ng of guinea pig pepsinogen, and the monolayer culture system responded well to secretagogues. These systems are useful for studying pepsinogen secretion.
Okayama
N
N
1st Department of Internal Medicine, Nagoya City University Medical School.
Itoh
M
M
Joh
T
T
Kasugai
K
K
Kawai
T
T
Yokoyama
Y
Y
Takeuchi
T
T
Katoh
T
T
Miyamoto
T
T
D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005753:Gastric Mucosa / Q000166:cytology / Q000378:metabolism*; D006168:Guinea Pigs; D007124:Immunoenzyme Techniques; D008297:Male; D010435:Pepsinogens / Q000187:drug effects / Q000378:metabolism*; D011817:Rabbits
D010435:Pepsinogens; D002217:Carbachol