A method for the isolation of human gastric mucous epithelial cells for primary cell culture: A comparison of biopsy vs surgical tissue

We have developed a method for the isolation and growth of normal human gastric mucous epithelial cells using biopsies or surgically resected tissues as the source of the cells. The attachment and growth of cells were dependent upon: (1) cell planting density, ∼50,000 cells/cm 2 ; (2) extracellular matrix (fibronectin); and (3) and the use of a porous filter. In all experiments we found better cells attachment and growth of human gastric mucous cells isolated from surgical specimens compared with those gastric mucous cells isolated from gastric biopsies. The initial cell viability (as measured by Trypan-blue) was the same in both populations of gastric mucous epithelial cells isolated from either gastric biopsies or surgical specimens. After 4–5 days in culture one could detect various amounts of mucin in all the cells using either periodic acid Schiff (PAS) staining or a specific anti-mucin antibody. A similar pattern of much straining was also found in primary cultures of guinea pig gastric mucous epithelial cells. Immunohistochemical staining for chief cells (anti-pepsinogen) or parietal cells (anti-H + /K + ATPasc) in the gastric mucous cuboidal-like epithelial cells with tight junctions, desmosomes,short microvilli, a filamentous terminal web, mucous granules, and basal lamina-like structure. We could not detect the presence of fibroblasts during the 7–9 days that the primary cells were in culture. This cell culture method will prove useful in the isolation of normal human gastric mucous epithelial cells for in vitro studies of gastric mucosal injury and repair.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/43235/1/11022_2004_Article_BF00127904.pd

Rapid effect of progesterone on transepithelial resistance of human fetal membranes: Evidence for non-genomic action

1. The factors that regulate human fetal membrane transport mechanisms are unknown. The aim of the present study was to investigate the effect of progesterone on transepithelial electrical resistance (R-TE) in the human amniochorion. 2. Fetal membranes from uncomplicated term pregnancies were obtained immediately after vaginal or Caesarean deliveries. Intact pieces were mounted as planar sheets separating an Ussing chamber. Progesterone (10(-4) to 10(-7) mol/L), mifepristone (10(-4) to 10(-8) mol/L) and combinations of progesterone plus mifepristone were applied to the chambers facing the fetal or maternal sides of the membrane. The R-TE was measured before and 1, 5, 10, 15, 20, 25, 30, 45 and 60 min after each solution was added (at 37 degrees C). The R-TE was calculated in Omega.cm(2), according to Ohm's law. 3. The mean (+/- SEM) basal value of R-TE before the application of any substance in all experiments was 29.1 +/- 0.4 Omega.cm(2). The net change in the R-TE (Delta R-TE) in relation to the basal value was calculated in each experiment. Progesterone, mifepristone and the combination of progesterone and mifepristone induced a rapid, surge-type increase in R-TE during the 1st min on both sides of the membrane. The combination of progesterone plus mifepristone exerted a synergistic action. The effect was stronger on the fetal side than on the maternal side for all substances tested (P < 0.05). The highest Delta R-TE during the 1st min on the fetal side was seen with the combination of progesterone plus mifepristone (4.0 +/- 0.3 Omega.cm(2)) and the lowest Delta R-TE occurred with mifepristone (1.5 +/- 0.1 Omega.cm(2)). 4. The present results demonstrated that the R-TE of human fetal membranes increases rapidly in response to progesterone. It is possible that changes in R-TE play a role in the control of membrane permeability during pregnancy