Generation and viability evaluation of human precision-cut placental slices for ex vivo studies.

INTRODUCTION

Exposure to medication during pregnancy can compromise placental function, impacting maternal and fetal health. This study aims to evaluate the viability of human precision-cut placental slices (hPCPSs) as a potential in vitro model, offering a new platform for experimental investigations in placenta.

METHODS

Decidual and villous hPCPSs of healthy, full-term placentas were prepared with a Krumdieck slicer and were cultured and analyzed at 0 h, 24 h, 48 h, and 72 h. Slice viability was assessed using ATP/protein content, lactate dehydrogenase (LDH) leakage and PrestoBlue assay. Progesterone, estradiol, estrone and human chorionic gonadotropin secretion were measured in culture supernatants. Morphological changes were microscopically evaluated via Hematoxylin and Eosin (H&E) staining. To evaluate the impact of delayed processing, slices from freshly processed placentas and stored at 4 °C for 24 h were also compared.

RESULTS

At 72 h, ATP decreased only in stored villous hPCPSs (P < 0.01), whereas PrestoBlue indicated reduced viability in all groups except fresh villous (all P < 0.05). LDH leakage peaked at 24 h across decidual and villous slices, irrespective of storage (P < 0.05). Hormone levels peaked at 24 h and subsequently decreased, with significance mainly in progesterone and estrone (P < 0.05). H&E showed progressive karyorrhexis and syncytiotrophoblast detachment across all groups.

CONCLUSION

Our studies established a reproducible protocol for generating hPCPSs, enabling separate evaluation of decidua and villi. Furthermore, our results underscore the importance of viability monitoring and suggest that hPCPSs maintain viability up to 48 h, even after short-term cold storage.