Isolation of parietal cells from guinea-pig gastric mucosa by a Percoll density gradient.
Isolation of parietal cells from guinea-pig gastric mucosa was performed by density gradient centrifugation using percoll, a new gradient medium composed of colloidal silica coated with polyvinylpyrolidone. The gastric mucosa of the oxyntic gland area was exfoliated, minced and digested with 2.0 mg/ml of dispase and 0.1 mg/ml of collagenase. Dispersed mucosal cells were applied to the percoll density gradient. Parietal cells were fractionated in the layer between 1.04 and 1.06 g/ml in density with purity 80 approximately 90%. For identification of parietal cells staining with tetranitrobluetetrazolium and electron microscopy were used in addition to common staining. The viability of the cells was over 95% assessed by trypan-blue exclusion. The parietal cells obtained responded actively to histamine stimulation when 14C-aminopyrine uptake was used as an functional index. In conclusion, percoll density gradient medium is suitable for yielding isolated parietal cells with both high purity and high viability.
D000632:Aminopyrine / Q000378:metabolism; D000818:Animals; D002469:Cell Separation; D002470:Cell Survival; D002499:Centrifugation, Density Gradient; D005753:Gastric Mucosa / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006168:Guinea Pigs; D006632:Histamine / Q000494:pharmacology; D008297:Male; D008854:Microscopy, Electron; D010295:Parietal Cells, Gastric / Q000166:cytology* / Q000378:metabolism; D011205:Povidone; D012822:Silicon Dioxide
D000632:Aminopyrine; C016039:Percoll; D012822:Silicon Dioxide; D006632:Histamine; D011205:Povidone