title: Effects of cell storage and passage on basal and oxytocin-regulated prostaglandin secretion by equine endometrial epithelial and stromal cells.
abstract: Cell cultures are useful for determining the responses of specific cell types to various factors under controlled conditions and for obtaining a better understanding of in vivo physiologic processes. The aims of the present study were (i) to establish methodologies for isolation, culture and cryopreservation of equine endometrial epithelial and stromal cells; and (ii) to determine the effect of passage and cryopreservation on endometrial cell physiology, based on their basal and oxytocin (OT)-stimulated prostaglandin (PG) release. Epithelial and stromal cells were obtained by enzymatic digestion of equine endometrium collected from Days 2-5 of the estrous cycle (n = 16). Primary epithelial and stromal cells, as well as cryopreserved cells were stimulated with OT (10(-7)m) for 24 h. The concentrations of PGE(2) and PGF(2α) in the culture medium were measured by enzyme-linked immunosorbent assay (EIA). Oxytocin increased PGE(2) and PGF(2α) release by primary cultures of unfrozen epithelial cells until passage I (P < 0.01) and by the primary culture of unfrozen and cryopreserved/thawed stromal cells until passage IV (P < 0.01). Cryopreserved/thawed stromal cells cultured up to passage IV and unfrozen epithelial cells derived from passage I have physiological properties similar to those observed in primary culture and may be successfully used for in vitro studies of PG secretion.
authors.affiliation: Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research, Olsztyn, Poland.
authors.forename: A Z
authors.initials: AZ
authors.lastname: Szóstek
authors.forename: M J
authors.initials: MJ
authors.lastname: Siemieniuch
authors.forename: A M
authors.initials: AM
authors.lastname: Galvão
authors.forename: K
authors.initials: K
authors.lastname: Lukasik
authors.forename: D
authors.initials: D
authors.lastname: Zieba
authors.forename: G M
authors.initials: GM
authors.lastname: Ferreira-Dias
authors.forename: D J
authors.initials: DJ
authors.lastname: Skarzynski
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000662:veterinary*; D015925:Cryopreservation / Q000662:veterinary; D004717:Endometrium / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005260:Female; D006736:Horses; D010121:Oxytocin / Q000494:pharmacology*; D011453:Prostaglandins / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
chemical_list: D011453:Prostaglandins; D010121:Oxytocin