Suppression of Apoptotic Cell Death Progressed in Vitro with Incubation of the Chorion Laeve Tissues of Human Fetal Membrane by Glucocorticoid.

Abstract

In the trophoblast layer of the chorion laeve of human fetal membranes obtained by cesarean section at the month of normal parturition, cells with condensed nuclei could be observed by histochemical examination. Incubating fetal membranes at 37 degrees C in vitro in cultivation medium, the frequency of cells with condensed nuclei increased in the chorion laeve, associating with an increase in DNA fragmentation and the population of in situ TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-X nick end labeling) staining-positive cells. The progressed apoptotic cell death in the chorion laeve in vitro was suppressed by incubation of the tissue in the presence of glucocorticoids, cortisone or hydrocortisone, which was also demonstrated by DNA fragmentation analysis and in situ TUNEL staining. These results reveal that a substantial proportion of trophoblast cells in the chorion laeve of human fetal membranes are induced to undergo apoptosis at the end of pregnancy, and that the apoptosis progresses rapidly in vitro as the incubation period increases. It is suggested that certain hormones such as glucocorticoid, may be related to the regulation of the apoptosis in human fetal membranes.

Cytotoxicity and Apoptotic Inducibility of Vitex agnus-castus Fruit Extract in Cultured Human Normal and Cancer Cells and Effect on Growth.

“…9) Extracted DNA was dispersed in a suitable volume of lysis buffer, and about 20 mg/20 ml of DNA was then loaded onto a 2% agarose (Agarose X, Nippon Gene, Tokyo, Japan) gel electrophoresis column. Gels were stained with ethidium bromide (Sigma Chemical Co.) and viewed under UV light.…”

Apoptosis in Cultured Human Fetal Membrane Cells Infected with Influenza Virus.

“…18) Extracted DNA was dissolved in TE buffer [10 mM Tris-HCl, pH 8.0, 1 mM ethylenediaminetetraacetic acid, disodium salt (EDTA-2Na)], and the DNA concentration was determined by staining with Hoechst 33258 as described. 19) Extracted DNA (10 mg) and a Ready-Load TM 100 bp DNA Ladder (GibcoBRL, MD, U.S.A.) as a DNA size marker were resolved by electrophoresis on 2.0% agarose gel (Agarose X, Wako Pure Chemical Industry, Japan) using TBE buffer (89 mM Tris, 89 mM boric acid, 2 mM EDTA-2Na).…”

Apoptosis Induced by Influenza Virus-Hemagglutinin Stimulation may be Related to Fluctuation of Cellular Oxidative Condition.

“…16) Confluence monolayer cells in a 25 cm 2 bottle were washed once with sterilized Hanks' balanced salt solution (GibcoBRL) and then inoculated with 1 ml of fractionated IV-HA solution. These cells were incubated at 37°C for 60 min rocking every 15 min.…”