Possible role of prostaglandin E2 in human amniotic epithelial cell death: an in vitro study.

Abstract

To understand the optimal conditions necessary for successful transplantation of normal human amniotic epithelial (HAE) cells in vivo, the factors which influence the cell death were investigated. In this study, the role of prostaglandin E2 (PGE2) in HAE cells in vitro was examined. Normal HAE cells. The HAE cells were incubated with PGE2 100 nM, 500 nM or 1,000 nM for 8 h. The PGE2 treated and untreated cells were collected at 2 h, 4 h and 8 h intervals and stained using trypan blue and terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL) methods to detect necrosis and apoptosis respectively. Prostaglandin E2 induced more HAE cell death at 8 h treatment. The percentages of cell death induced by PGE2 100 nM, 500 nM and 1,000 nM at 8 h were 14.0 +/- 0.35 (n=9), 14.9 +/- 0.64 (n=9) and 18.3 +/- 0.65 (n=9) respectively compared to the control (7.0 +/- 0.35; n = 9) (P < 0.001) determined using trypan blue staining method. The apoptotic cell death induced by the above doses of PGE2 at 8 h were 5.7 +/- 0.6 (n=9), 7.14 +/- 0.84 (n=9) and 8.6 +/- 0.67 (n=9) respectively compared to the control (0.86 +/- 0.05; n=9) (P<0.001) determined using TUNEL method. The PGE2 induced high level cell death was via necrosis rather than apoptosis. These results indicate that PGE2 is an important mediator in HAE cell death.