Establishment and characterization of female reproductive tract epithelial cell culture

Abstract

The oviductal and uterine epithelial cells have a crucial role, but are still poorly understood. Numerous studies have tried to isolate the epithelial cells from different organs in various models. The current study aimed to establish and characterize an in vitro monolayer culture of the oviduct and uterine horn epithelial cells by using two different techniques. Female reproductive epithelial cells from sows were cultured in follicular phase. Combined protocols to isolate the epithelial cells were performed. The viability and cell number were determined. Monolayers of epithelial cells from each group were cultured in four-well plates and were subjected to immunostaining using a Vector ABC Elite Kit. The immunohistochemical staining step was performed to evaluate the quality of the epithelial cells. Oviductal cells reached confluence faster than uterine horn cells. Cilia were seen in oviduct and uterine horn tissue culture. All the isolated cells reached confluence prior to harvesting. The number of cells was increased over the time of incubation. Monolayer culture using the trypsin/EDTA method took longer than culture with the collagenase method. Immunohistochemistry of epithelial cells showed strong staining for cytokeratin. Oviductal and uterus epithelial cells were cultured and established. Both techniques used in this experiment were useful and showed no significant differences. This cell culture model has the potential to study the secretory interactions of the female reproductive tract with spermatozoa, oocytesor embryos.