Organ culture of amniochorionic membrane in vitro.

PROBLEM: The purpose of the study was to develop a novel method of amniochorionic membrane culture aimed at maintaining tissue integrity.

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.