Kisspeptin-1 receptor (Kiss-1R) expression has been reported in the placenta and a possible involvement of the Kiss-1R/Kisspeptins (Kiss-1R/Kps) system in regulating trophoblast invasion and proliferation has been hypothesized. The aim of the present study was to investigate whether Kiss-1R activation by Kisspeptin 10 (Kp-10) could modulate in vitro proliferation and progesterone (P4) secretion of bovine primary placental cell lines isolated from cotyledons of foetuses in the first pregnancy trimester. The involvement of Kiss-1R in observed cell responses was also analyzed. Uteri from cows at the first trimester of pregnancy were obtained from local abattoirs. Fetal cotyledon fragments were digested with collagenase in Low Glucose Dulbecco's Modified Eagle's Medium (LG-DMEM) and cells were collected and cultured in a LG-DMEM based expansion medium in presence of 0.01, 0.1, 1 and 10 µM Kp-10. Control cells were cultured in the absence of Kp-10. The total number of living cells was obtained every 3 days for each culture passage (P) from P1 to P10. Cells were tested for Kiss-1R mRNA expression analysis by Real Time RT-PCR and culture media were analyzed for P4 concentration by radioimmunoassay. Kp-10 modulated in vitro proliferation and P4 secretory activity of epithelial cell lines isolated from cotyledons recovered from bovine fetuses in the first trimester of pregnancy. Markedly different effects, inhibitory or stimulatory, of Kp-10 on cell proliferation were found in different cell lines and observed cell responses were found as being related to Kiss-1R mRNA levels and P4 secretion. Inhibition of cell proliferation matched with not significant variation of Kiss-1R expression but increased P4 secretion, whereas stimulation of cell proliferation was found as being related to Kiss-1R up-regulation and decreased P4 levels. These results allow to hypothesize that the Kiss-1R/Kps system could play a role in the fine developmental regulation of the bovine placental cotyledonal cell proliferation and P4 secretion in the first trimester of pregnancy.
Effects of Kisspeptin-10 on in vitro proliferation and Kisspeptin receptor(Kiss-1R) expression in primary epithelial cell cultures isolated from bovine placental cotyledons of embryos/fetuses at the first trimester of pregnancy
F Pizzi;
2014
Abstract
Kisspeptin-1 receptor (Kiss-1R) expression has been reported in the placenta and a possible involvement of the Kiss-1R/Kisspeptins (Kiss-1R/Kps) system in regulating trophoblast invasion and proliferation has been hypothesized. The aim of the present study was to investigate whether Kiss-1R activation by Kisspeptin 10 (Kp-10) could modulate in vitro proliferation and progesterone (P4) secretion of bovine primary placental cell lines isolated from cotyledons of foetuses in the first pregnancy trimester. The involvement of Kiss-1R in observed cell responses was also analyzed. Uteri from cows at the first trimester of pregnancy were obtained from local abattoirs. Fetal cotyledon fragments were digested with collagenase in Low Glucose Dulbecco's Modified Eagle's Medium (LG-DMEM) and cells were collected and cultured in a LG-DMEM based expansion medium in presence of 0.01, 0.1, 1 and 10 µM Kp-10. Control cells were cultured in the absence of Kp-10. The total number of living cells was obtained every 3 days for each culture passage (P) from P1 to P10. Cells were tested for Kiss-1R mRNA expression analysis by Real Time RT-PCR and culture media were analyzed for P4 concentration by radioimmunoassay. Kp-10 modulated in vitro proliferation and P4 secretory activity of epithelial cell lines isolated from cotyledons recovered from bovine fetuses in the first trimester of pregnancy. Markedly different effects, inhibitory or stimulatory, of Kp-10 on cell proliferation were found in different cell lines and observed cell responses were found as being related to Kiss-1R mRNA levels and P4 secretion. Inhibition of cell proliferation matched with not significant variation of Kiss-1R expression but increased P4 secretion, whereas stimulation of cell proliferation was found as being related to Kiss-1R up-regulation and decreased P4 levels. These results allow to hypothesize that the Kiss-1R/Kps system could play a role in the fine developmental regulation of the bovine placental cotyledonal cell proliferation and P4 secretion in the first trimester of pregnancy.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
