Steroid metabolism by cells from human chorion laeve isolated on Percoll gradients.
Collagenase-dispersed cells from human chorion laeve were examined on Percoll gradients. The 3 beta-hydroxysteroid dehydrogenase (a trophoblast marker) and steroid sulfatase activities of the cells were measured and a system was developed to isolate enriched preparations of the trophoblast cells. No cells were found to sediment at Percoll concentrations greater than 50%, and using continuous gradients of Percoll there appeared to be cells with different 3 beta-hydroxysteroid dehydrogenase (3 beta HSD): steroid sulfatase ratios sedimenting in different regions of the gradient. Cells with a high ratio were found in the denser region of the gradient. Continuous gradients provided inadequate separations of distinct populations of cells, thus to obtain a more reproducible system to isolate cells, discontinuous gradients of Percoll were studied. A discontinuous gradient composed of 5, 20, 40, and 60% Percoll was developed and three bands of cells were found sedimenting at the 20, 40 and 60% interfaces, respectively. The number and appearance of cells at the 20 and 60% interfaces varied from tissue to tissue. In contrast, the cells sedimenting at the 40% interface were less variable, a substantial number was found to be present in every tissue studied, they were similar in appearance to the trophoblast cells and had high 3 beta HSD:sulfatase ratios.
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D002469:Cell Separation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D002823:Chorion / Q000166:cytology / Q000378:metabolism*; D005260:Female; D006801:Humans; D011205:Povidone; D011247:Pregnancy; D012822:Silicon Dioxide; D013256:Steroids / Q000378:metabolism*; D013429:Sulfatases / Q000378:metabolism; D014048:Tolonium Chloride
D013256:Steroids; D014048:Tolonium Chloride; C016039:Percoll; D012822:Silicon Dioxide; D015096:3-Hydroxysteroid Dehydrogenases; D013429:Sulfatases; C038087:steroid 21-sulfatase; D011205:Povidone