An In Vitro Culture Model of Bovine Oviduct Epithelial Cells on Collagen Hydrogels

MILAGROS PEÑA; GARCIA, DANIELA CELESTE; EUGENIA MARIELA ROLDÁN-OLARTE; PABLO ALBERTO VALDECANTOS

Buenos Aires

Congreso; Reunión Conjunta de Sociedades de Biociencias; 2017

Sociedades de Biociencias

The epithelium lining of the mammalian oviduct has critical roles in the female fertility facilitating the gamete transport, oocyte fertilization, and early embryo development. Ciliated and non-ciliated secretory cells are the two major cell types of the oviduct epithelium. Different in vitro systems for the culture of oviduct epithelial cells (OEC) have been established; however, OEC grown on solid supports dedifferentiate into non-polarized cells; free-floating OEC-vesicles with active cilia on their external surface can be cultured for short terms; polarized OEC can be obtained for long term periods on both, permeable membrane inserts at air-liquid interfaces, and in Matrigel with cells orientated with the apical side into the organoid´s lumen. The aim of this work was to establish and characterize an in vitro culture of functional polarized bovine oviduct epithelial cells (BOEC) grown at an air-liquid interface on free-floating collagen hydrogels disc (10 mm x 2 mm). Type I collagen was extracted from rat tail tendons and sterile collagen hydrogels (2 mg/mL) were obtained. BOEC sheets free of stromal cells were obtained by gently squeezing of the oviducts. Epithelial sheets were immediately cultured on the top of collagen hydrogels submerged in DMEM supplemented with 10% Fetal Bovine Serum. After 48 h, cells adhered to the hydrogel surface; then, the collagen gels were maintained with the top surface above of the culture medium in an air-liquid interphase. Cultures were controlled under an inverted microscope for the presence of active cilia beating with medium changes at every 48 h. After three weeks, epithelial 3-D cultures were processed for histochemical staining and RT-PCR. Both, the epithelial phenotype mimicking the oviduct epithelium lining, and the expression of the oviduct specific glycoprotein 1 (OVGP1) are indicative of the potential of this culture system for being used as a model for the in vitro study of the oviduct epithelial function.