Cortisol in Fetal Fluids and the Fetal Adrenal: MATERIALS AND METHODS(2)

Cortisol Production

Adrenals from fetuses between Day 23 and Day 26 of gestation were placed in 24-well culture dishes in 1 ml of Medium 199 (Sigma, St. Louis, MO) and were sealed in an incubation chamber, gassed with 95% O2:5% CO2, and maintained at 37°C.Tissueswere allowedto equilibrate for 1 h before pretreatment samples (100 ^l) were taken. Adrenal pairs (left and right) were divided and randomly assigned to control or treatment groups. One adrenal always served as a control (no treatment), and the other received either 100 ng/ml synthetic ACTH (Synacthen; Ciba-Geigy, Pendle Hill, NSW, Australia) or 100 ng/ml PGE2 (Sigma). After treatment, the incubation chamber was re-gassed and returned to 37°C.Final sanples(200 |xl) were taken after 24 h and stored at -20°CunSllaseayed.Fenaladrenaletreaal ed with ACTH were from Day 24.4 ± 1.0 (n = 10) and those treated with PGF2 were from Day 24.7 ± 0.9 (n = 4). Cheap Diskus Advair

Cortisol Assay

netal and neonatal adrenal glands were thawed and homogenized in 12 X 75-mm glass tubes in 100 ^l 1.0 M NaOH using a glass stirring rod. The homogenate was neutralized with 100 ^l 1.0 M HCl, and a 10-^l aliquot was assayed for protein content, since significant dehydration of the glands occurs after freezing because of their minute size of less than 0.7 mg each. Cortisol was determined by RIA using a double-antibody, [125I]cortisol assay kit from Pantex (Santa Monica, CA).

This entry was posted in Adrenal and tagged Adrenal, Cortisol, Fetal Fluids, Parturition.