PROBLEM:
The purpose of the study was to develop a novel
method of amniochorionic membrane culture aimed at
maintaining tissue integrity.
METHOD:
METHOD: Amniochorionic
membranes were collected from women prior to labor, undergoing
elective cesarean section, with no history of infection or
pregnancy related complication. Fetal membranes were
maintained in culture for up to ten days.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a "house
keeping" gene and inflammatory cytokine mRNA was detected
by reverse transcriptase-polymerase chain reaction (RT-PCR) at
various periods of culture. mRNA localization was performed by
in situ hybridization.
RESULTS:
GAPDH gene expression was
seen throughout the culture period. Inflammatory cytokine
mRNA (IL-1 alpha, IL-1 beta and IL-6) were also detected
during culture. Cellular and tissue morphology appeared normal.
CONCLUSIONS:
The culture technique we propose is a simple
organ explant system which maintains the morphology and
autocrine/paracrine relationships within this tissue.